,, 曹梦竹, 游义琳, 黄卫东中国农业大学食品科学与营养工程学院/北京市葡萄科学与酿酒技术重点实验室,北京 100083Research Advance on the Application of Non-Saccharomyces in Winemaking
ZHAN JiCheng,, CAO MengZhu, YOU YiLin, HUANG WeiDongCollege of Food Science and Nutritional Engineering, China Agricultural University/Beijing Key Laboratory of Viticulture and Enology, Beijing 100083通讯作者:
责任编辑: 赵伶俐
收稿日期:2019-12-20接受日期:2020-02-20网络出版日期:2020-10-01
| 基金资助: |
Received:2019-12-20Accepted:2020-02-20Online:2020-10-01
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战吉宬, 曹梦竹, 游义琳, 黄卫东. 非酿酒酵母在葡萄酒酿造中的应用[J]. 中国农业科学, 2020, 53(19): 4057-4069 doi:10.3864/j.issn.0578-1752.2020.19.018
ZHAN JiCheng, CAO MengZhu, YOU YiLin, HUANG WeiDong.
考古证据表明,葡萄酒人工酿造已有8 000多年的悠久历史。葡萄酒的酿造离不开酵母。从葡萄园和酒厂分离得到的酵母中除酵母属（Saccharomyces）（又常称酿酒酵母属）外均为非酿酒酵母。葡萄酒酿造中常见的酵母属酵母有4个种：酿酒酵母Saccharomyces cerevisiae、贝酵母Saccharomyces bayanus（原葡萄汁酵母Saccharomyces uvarum）、巴斯德酵母Saccharomyces pastorianus和奇异酿酒酵母Saccharomyces paradoxus[1]。其中酿酒酵母是主导酒精发酵最重要的菌株,能够将糖转化成酒精和二氧化碳,并通过二级代谢产生风味物质,实现快速、完全、高效的发酵。
非酿酒酵母（non-Saccharomyces）因常从发酵停滞或变质的酒及果实中分离到,并与乙酸、乙醛、硫化氢等异味物质的产生有关,早期被认为是有害菌株[2]。但随着研究者们对酿酒酵母的深入研究发现,在酿酒过程中可以利用非酿酒酵母来改善葡萄酒的风味物质组成,如增加甘油、总酸、挥发性酯类含量,降低乙酸含量,形成稳定的吡喃花青素等,进而改善酒的整体品质[3]。葡萄酒中常见的非酿酒酵母属主要包括有孢圆酵母属（Torulaspora）、有孢汉逊酵母属（Hanseniaspora）、假丝酵母属（Candida）、接合酵母属（Zygosaccharomyces）、毕赤酵母属(Pichia) 、裂殖酵母属（Schizosaccharomyces）、美奇酵母属（Metschnikowia）、类酵母属（Saccharomycodes）、伊萨酵母属（Issatchenkia）、隐球酵母属（Cryptococcus）、酒香酵母属（Brettanomyces）、德巴利酵母属（Debaryomyces）、克鲁维酵母属（Kluyveromyces）、克勒克酵母属（Kloeckera）、红酵母属（Rhodotorula）、德克酵母属（Dekkera）、汉逊酵母属（Hansenula）、洛德酵母属（Lodderomyces）、拟威尔酵母属（Williopsis）、固囊酵母属（Citeromyces）、接合囊酵母属（Zygoascus）、Kazachstania等[4,5]。需要注意的是,酵母分有性繁殖和无性繁殖两种繁殖方式,在葡萄酒酿造中绝大部分以无性繁殖为主。非酿酒酵母名称繁多,有性型和无性型通常对应不同属种。KURTZMAN[5]对此进行了总结,对应关系见表1。
Table 1
表1
表1部分非酿酒酵母有性型、无性型和别名[5]
Table 1
| 有性型 Anamorphic | 无性型 Teleomorphic | 别名 Synonym |
|---|---|---|
| 高糖固囊酵母 Citeromyces matritensis | 球形假丝酵母 Candida globosa | |
| 耐热克鲁维酵母 Kluyveromyces thermotolerans | Lachancea thermotolerans | |
| 布鲁塞尔德克酵母Dekkera bruxellensis | 布鲁塞尔酒香酵母Brettanomyces bruxellensis | |
| 季也蒙有孢汉逊酵母Hanseniaspora guilliermondii | 蜜蜂克勒克酵母Kloeckera apis | |
| 罕见有孢汉逊酵母Hanseniaspora occidentalis | 日本克勒克酵母Kloeckera javanica | |
| 嗜高压有孢汉逊酵母Hanseniaspora osmophila | 皮质克勒克酵母Kloeckera cortices | |
| 葡萄汁有孢汉逊酵母Hanseniaspora uvarum | 柠檬形克勒克酵母Kloeckera apiculata | |
| 葡萄酒有孢汉逊酵母Hanseniaspora vineae | 非洲克勒克酵母Kloeckera Africana | |
| 美极梅奇酵母Metschnikowia pulcherrima | 铁红假丝酵母Candida pulcherrima | Torulopsis pulcherrima |
| 季也蒙毕赤酵母Pichia guilliermondii | 季也蒙假丝酵母Candida guilliermondii | |
| 发酵毕赤酵母Pichia fermentans | 郎比可假丝酵母Candida lambica | |
| 膜醭毕赤酵母Pichia membranifaciens | 粗壮假丝酵母Candida valida | |
| 东方伊萨酵母Issatchenkia occidentalis | Candida sorbosa | |
| 陆生伊萨酵母Issatchenkia terricola | 陆生毕赤酵母Pichia terricola | |
| 戴尔有孢圆酵母Torulaspora delbrueckii | 软假丝酵母Candida colliculosa | Saccharomyces rosei |
| 异常毕赤酵母Pichia anomala | 菌膜假丝酵母Candida pelliculosa | 异常威克汉姆酵母Wickerhamomyces anomalus/异常汉逊酵母Hansenula anomala |
| Candida zemplinina | Starmerella bacillaris | |
| 星型假丝酵母Candida stellata | 星形球拟酵母Torulopsis stellata |
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多数研究结果表明,葡萄酒自然发酵初期由克勒克酵母、有孢汉逊酵母和假丝酵母等主导发酵,细胞浓度甚至可达106—108 CFU/mL,中期发现有美奇酵母和毕赤酵母,末期可检测到酒香酵母、克鲁维酵母、裂殖酵母、有孢圆酵母、接合酵母等[6]。虽然因产地和浆果状态不同,不同文献结果有所差异,但均说明酒精发酵过程中非酿酒酵母广泛存在且数量不低。一些品酒师认为自然发酵酒比目前广泛使用的商业活性干酵母（酿酒酵母）接种发酵酒具有更加优良的风味特征。研究人员因此开始关注和评估非酿酒酵母在葡萄酒发酵中的作用。由于具有优良酿酒特性的转基因酿酒酵母不允许被商业化使用,而非酿酒酵母的使用不涉及监管、法律和道德问题,因此非酿酒酵母单独发酵或和酿酒酵母混合发酵成为一条目前可行的改善葡萄酒风味的途径。不仅如此,有研究还表明一些非酿酒酵母的参与有助于保障葡萄酒安全[7]。
1 非酿酒酵母的酿造特性
葡萄酒发酵是一个复杂的微生物反应过程,葡萄醪的理化条件和其中的微生物相互作用,进而影响酒的品质。目前葡萄酒行业商业非酿酒酵母的使用多以混合发酵方式进行,即酿酒酵母和非酿酒酵母混合发酵的方式,包括同时接种发酵和顺序接种发酵。非酿酒酵母的使用不要求其持续存在于发酵过程中,可以只利用其在发酵前期的一些功能,如水解芳香化合物前体,合成酯类、醇类、甘油等风味物质,后期发酵可任其自然消亡,由酿酒酵母主导完成发酵。非酿酒酵母具有独特的代谢通路和较强的酶活,可以在降低酒精生成的同时增加甘油、萜烯和酯类含量,释放甘露糖蛋白或多糖改善口感,增强颜色稳定性[7]。不同属种的非酿酒酵母具体贡献不同,了解各个属种的发酵特性、根据发酵目的选择合适的接种菌株极为重要。1.1 有孢圆酵母属（Torulaspora）
有孢圆酵母属包括戴尔有孢圆酵母（Torulaspora delbrueckii）、T. franciscae、T. globosa、T. maleeae、T. microellipsoides和T. pretoriensis等。公认的对葡萄酒感官特征有积极影响的T. delbrueckii是目前应用最多、商业化程度最高的非酿酒酵母菌种。已实现商业化的有Prelude?（http://www.chr-hansen.com）、Biodiva?（http://www.lallemandwine.com）、Zymaflore?Alpha（https://www.laffort.com/es/productos/levaduras-zymaflore）、Vinifer NS TD（http://www.agrovin.com）和PrimaFLLA?VB BIO（http://www.sud-et-bio.com）。T. delbrueckii发酵能力较强,纯种发酵可获得7.4%—9.38%（v/v）酒精度,耐酒精和高渗透压,高产果味脂肪酸乙酯如丙酸乙酯、辛酸乙酯、癸酸乙酯、甘油、萜烯和硫醇,低产高级醇、挥发酸和乙醛,能释放较多的甘露糖蛋白和多糖,对花色苷吸收较少。可能的副作用包括少量4-乙基苯酚的产生,较高的琥珀酸浓度和组氨酸（生物胺前体）的轻微增加[8]。T. delbrueckii适合高糖分葡萄如晚收葡萄和冰酒的发酵,也可用于起泡基酒的发酵[9];而T. delbrueckii和酿酒酵母混合发酵则可降低酒精含量,降低60%乙醛和0.08—0.27 g·L-1乙酸,降解20%—25%苹果酸,降低70—90 mg·L-1高级醇,但可提高0.1—1 g·L-1甘油产量,增加2-苯乙醇和2-苯乙酸乙酯、乳酸乙酯、乙酸异戊酯、乙酸己酯、己酸乙酯和辛酸乙酯等乙酯类化合物浓度[10-13] ,释放较高浓度的萜烯如α-松油醇、芳樟醇、香叶醇[14],增加100—300 ng·L-1 3-巯基己醇（3MH）和70 ng·L-1 4-巯基-4-甲基-2-戊酮（4MMP）等硫醇含量[15],提高芳香型葡萄品种如琼瑶浆（Gewurztraminer）、雷司令（Riesling）葡萄酒的品种香气,增加花色苷总量和颜色强度[16],提高起泡酒发泡性和泡沫持久性[17]。概括来说,T. delbrueckii与酿酒酵母混合发酵可使酿成的葡萄酒花香和果香增强,植物性香气降低,香气复杂性增加,有时甚至会出现少见的干果/油酥糕点香气。值得注意的是,不同文献的报道结果会出现矛盾,这主要与菌株特异性和发酵条件特异性等有关。菌株特异性在所有属种的发酵中都非常常见,表明菌株选择具有重要意义。发酵条件的特异性则表明具体应用前尚需小试才行。1.2 有孢汉逊酵母属（Hanseniaspora）
有孢汉逊酵母属主要包括葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）、葡萄酒有孢汉逊酵母（Hanseniaspora vineae）、季也蒙有孢汉逊酵母（Hanseniaspora guilliermondii）、仙人掌有孢汉逊酵母（Hanseniaspora opuntiae）、嗜高压有孢汉逊酵母（Hanseniaspora osmophila）、罕见有孢汉逊酵母（Hanseniaspora occidentalis）等。目前上述各种多有研究,相比酿酒酵母纯种发酵,H. uvarum纯种发酵产生更低的挥发酸,更高的甘油、有机酸、醛和次级醇,感官品评得分更高[18]。与酿酒酵母同时接种混合发酵则能显著提高C-13降异戊二烯、萜烯和乙酯的含量,添加胞外提取物也有类似的效果,能显著提高品种香气[19]。顺序接种丹麦葡萄品种Solaris则产生较高水平的甘油、乙酸庚酯和2-苯乙酸乙酯[20]。另外,TRISTEZZA等[21]在实验室、小试和工厂水平都发现混合接种有助于提高葡萄酒感官品质,降低挥发酸。但顺序接种过量的H. uvarum也会提高乙酸酯和挥发酚浓度,给赤霞珠（Cabernet Sauvignon）酒带来指甲油气味;H. vineae和酿酒酵母顺序接种的葡萄酒更干（发酵更彻底）,甘油、乙酰、乙酯含量显著增加,醇和脂肪酸含量减少,香气复杂性增加,香蕉、梨、柠檬、番石榴等果味强度显著提高[22]。LLEIXA等[23]还发现H. vineae能够在马卡贝奥（Macabeo）葡萄果实发酵中占据主导地位,酿造出更富含果香和花香的葡萄酒,苯乙酸乙酯浓度可达酿酒酵母的50倍;H. guilliermondii和酿酒酵母混合发酵可显著增加2-苯乙醇和2-苯乙酸乙酯含量,赋予葡萄酒果香与花香。2-苯乙醇可达6.3 mg·L-1（酿酒酵母纯种发酵不超过1.12 mg·L-1）,2-乙酸苯乙酯可达11.1 mg·L-1（酿酒酵母纯种发酵仅可产生0.22—0.25 mg·L-1）[24];H. opuntiae和酿酒酵母同时接种增加了88.61%的酯类和21.40%的萜烯,乙酸苯乙酯增产14.60倍,β-大马士酮增产8.85%,感官品评果香、花香和黄油属性显著增强[25];H. osmophila和酿酒酵母混合发酵的2-苯乙酸乙酯为酿酒酵母纯种发酵的3—9倍,感官品评果味更强,乙酸和乙酸乙酯浓度在合理范围内[26]。从各种应用研究的结果来看,该属发酵能力和耐酒精能力较差,无法独立完成发酵,主要在发酵前期占主导地位,进入中后期会逐渐凋亡。优点是发酵过程中能产生大量带有果香和花香属性的乙酸酯,特别是2-苯乙酸乙酯,使香气的复杂性增加;可能的副作用是产生过量的乙酸乙酯。1.3 假丝酵母属（Candida）
假丝酵母属已经鉴定的种很多,在KURTZMAN等[5] 2011版的书中记录了314种。葡萄酒应用研究中发现的主要包括星形假丝酵母（Candida stellata）、Candida zemplinina、铁红假丝酵母（Candida pulcherrima）、季也蒙假丝酵母（Candida guilliermondii）等。其中,C. stellata可耐受10%—12%乙醇,高产甘油（高达10—14 g·L-1,酿酒酵母只产生4—7 g·L-1）、琥珀酸和酯,低产乙酸和高级醇。可能的副作用为高产2,3-丁二醇,过浓的黄油、奶酪味道会掩盖果香;C. zemplinina在葡萄酒发酵过程中也比较常见。分子分类学检查发现41株原描述为C. stellata的菌株大部分实际为C. zemplinina及其相关种[27]。C. zemplinina和酿酒酵母混合发酵的最大特征是高甘油和低乙醇含量[28]。ENGLEZOS等[29]也发现C. zemplinina和酿酒酵母混合发酵能产生更多的甘油和令人愉悦的酯,可以改善巴贝拉（Barbera）葡萄酒的口感和香气特征。RANTSIOU等[30]发现混合发酵在降低乙酸含量的同时保持高水平的甘油和乙醇产量,建议用于甜酒生产;C. pulcherrima与酿酒酵母混合发酵可促进酒精、酯类和萜类的生成,果香和花香表现最好[31]。总的来说,假丝酵母为好氧型酵母,发酵初期可大量存活,发酵能力和耐酒精能力较强,显著特点是高产甘油。1.4 接合酵母属（Zygosaccharomyces）
接合酵母属主要包括拜尔接合酵母（Zygosaccharomyces bailii）、Zygosaccharomyces florentinus、发酵型接合酵母（Zygosaccharomyces fermentati）、鲁氏接合酵母（Zygosaccharomyces rouxii）、Zygosaccharomyces kombuchaensis等。研究发现,Z. bailii混合发酵霞多丽（Chardonnay）能产生高水平乙酯,带有令人愉悦的果香和花香,还可以高效分解苹果酸[32];Z. fermentati混合发酵桑娇维塞（Sangiovese）,实验室规模下多糖和2-苯乙醇含量增加,挥发酸浓度居于二者纯种发酵之间;酒厂规模下顺序接种显著增加甘油和酯类含量,感官品评花香较高,收敛性较低[33];Z. kombuchaensis则可以提高丽波拉（Ribolla）葡萄酒的香气强度和水果特性[34]。从上述研究结果看,接合酵母具有增加果香和花香的能力,部分接合酵母还可以降解苹果酸,可用于发酵降酸。1.5 毕赤酵母属（Pichia）
毕赤酵母属主要包括发酵毕赤酵母（Pichia fermentans）、异常毕赤酵母（Pichia anomala）、膜璞毕赤酵母（Pichia membranifaciens）、库德里阿兹威氏毕赤酵母（Pichia kudriavzevii）、季也蒙毕赤酵母（Pichia guilliermondii）等。研究发现,相比酿酒酵母纯种发酵,P. fermentans和酿酒酵母混合发酵有较强的增香酿造潜力,可提高26%的品种香气、39%发酵香气,提高40%中链脂肪酸乙酯含量[35]。MA等[36]发现混合发酵过程中C2—C8酯酶活性更高,可显著增加乙酸酯、乙酯、脂肪酸和其他挥发性物质含量,特别是中链脂肪酸及其相应酯的含量。添加P. fermentans胞外酶也可显著增加多种芳香化合物的释放,如萜烯、C13-降异戊二烯和C6化合物,感官分析具有浓郁的酸甜果香。CANAS等[37]发现P. anomala和酿酒酵母顺序接种佳利酿（Carignan）增加了乙酸酯、乙酯和直链醇浓度,有机酸浓度降低,感官品评果香和花香更浓且更受欢迎。总的来讲,毕赤酵母对风味贡献主要是影响酯类含量,具有增香潜力,能够改善葡萄酒的果香和花香。1.6 裂殖酵母属（Schizosaccharomyces）
裂殖酵母属中研究比较多的主要是粟酒裂殖酵母（Schizosaccharomyces pombe）和日本裂殖酵母（Schizosaccharomyces japonicas）等。2013年,裂殖酵母被OIV同意使用。发酵能力与酿酒酵母相当,但易产生异味物质。S. pombe最显著的特征是能通过丙二醇发酵,近100%降解苹果酸,适合寒冷气候产区的降酸发酵[12]。可将糖类高效转化为酒精,提高丙酮酸和乙烯基酚类吡喃花青素含量,增加多糖释放,降低尿素等生物胺前体含量,适合生产具有增强口感和降低酸度的天然改性葡萄酒。可能的副作用包括高产乙酸（高达1 g?L-1）和硫化氢[38]。S. pombe纯种发酵和混合发酵均可使苹果酸浓度从初始5.5 g·L-1降至小于0.5 g·L-1,乙酸浓度小于0.4 g·L-1（此处存在菌株特异性）。混合发酵酒中尿素含量降低10倍,感官品评发酵香气强度最大且更受欢迎[39]。IVIT等[40]发现瓶内带渣二次发酵起泡红葡萄酒增加了吡喃花色苷含量和色度。DOMIZIO等[41]发现酒精发酵过程中S. pombe释放的多糖为酿酒酵母的3—7倍,S. japonicas释放的多糖含量更高,同时也证实了高水平丙酮酸的存在。S. japonicas的混合发酵也对最终乙醇浓度、降低总酸、增加挥发性化合物和多糖浓度有重大贡献,具有良好的应用前景[42]。该属目前最受到关注的点是其降酸功能,寒冷气候区降酸困难时可以考虑尝试一下。1.7 梅奇酵母属（Metschnikowia）
梅奇酵母属中与葡萄酒酿造最密切相关的是美极梅奇酵母（Metschnikowia pulcherrima）,具有高β-葡萄糖苷酶活性。纯种发酵能够增加α-松油醇、橙花醇和香叶醇含量,降低乙醇、乙酸和硫化氢产量[43]。混合发酵可降低马卡贝奥起泡酒挥发酸浓度,增加中链脂肪酸、高级醇、酯、萜烯和甘油产量,提升泡沫持久性和烟熏、花香等香气性[44],提高雷司令葡萄酒中的柑橘、葡萄、梨等果香和总体得分[45]。同时接种美乐（Merlot）可降低1.0%酒精,总酯、总高级醇和总硫化合物浓度较高,感官品评红果和果香得分较高[46],可能的副作用为与酿酒酵母拮抗导致发酵延迟[7]。1.8 德巴利酵母属（Debaryomyces）
德巴利酵母属主要包括Debaryomyces vanriji和Debaryomyces pseudopolymorphus等。具有高β-葡萄糖苷酶活性,能够增加萜烯含量[47]。接种D. vanriji发酵的酒富含脂肪酸、酯类和萜烯,发酵过程中β-葡萄糖苷酶活性更高[48],和酿酒酵母混合发酵佩德罗-吉梅内斯（Pedro Gimenez,阿根廷白葡萄品种）也增加了酯和脂肪酸浓度[47]。1.9 伊萨酵母属（Issatchenkia）
伊萨酵母属主要包括东方伊萨酵母（Issatchenkia orientalis）和陆生伊萨酵母（Issatchenkia terricola）等,具有良好的苹果酸降解能力。HONG等[49]使用I. orientalis处理含有高水平苹果酸的葡萄酒,苹果酸含量由8.96 mg·mL-1降至0.75 mg·mL-1,降解率为91.6%。混合发酵有效提高了香气复杂性,虽然醇类含量相比酿酒酵母纯种发酵减少8%,但酯类含量增加30%,醇类和酯类种类也增加[50]。I. terricola在适当条件下可以同时降解苹果酸和柠檬酸,降解率均达90%以上[51]。该属也是降酸实践比较理想的选择。1.10 克鲁维酵母属（Kluyveromyces）
克鲁维酵母主要包括耐热克鲁维酵母（Kluyveromyces thermotolerans）、乳酸克鲁维酵母（Kluyveromyces lactis）、马克思克鲁维酵母（Kluyveromyces marxianus）等。与葡萄酒最密切相关的是K. thermotolerans,高产乳酸（纯种发酵可达9.6 g·L-1）,低产醋酸和异味,适合在温带葡萄栽培区酸化葡萄酒使用[52]。混合接种可显著降低pH,降低挥发酸,提高总酸,增加甘油和2-苯乙醇含量,感官品评显示香辛料香气和总酸显著增加[53]。也可以将S. pombe和K. thermotolerans共同使用,降解苹果酸,产生乳酸,所酿酒果味特征更加明显,乙酸和生物胺含量更低,这种新的红葡萄酒酿造技术有望取代传统苹乳发酵[54]。1.11 隐球酵母属（Cryptococcus）
隐球酵母属中与葡萄酒最密切相关的是浅黄隐球酵母（Cryptococcus flavescents）,混合发酵可提高蛇龙珠（Cabernet Gernischt）葡萄酒的香气,相比酿酒酵母纯种发酵,总醇、总酸、总酯含量可增加2倍多,萜烯含量增加4倍,且只在混合发酵中存在异戊醇、2-庚醇、丙酸乙酯、己酸乙酯、乙酸丙酯、β-大马酮等物质[55]。1.12 类酵母属（Saccharomycodes）
类酵母属中与葡萄酒最密切相关的是路德类酵母（Saccharomycodes ludwigii）,常在发酵结束时或葡萄酒储存期间被分离,耐高酒精、高SO2、高产多糖和芳香族化合物等物质。PALOMERO等[56]发现S. ludwigii在发酵过程中比酿酒酵母多释放300%多糖,酒糟陈酿过程中多释放200%多糖。相比酿酒酵母纯种发酵,混合发酵可降低酒精含量,增加芳香族化合物产量。如DOMIZIO等[57]发现混合发酵降低了1.74%酒精,增加21.8%甘油、20.8%乙酸异戊酯、200% 2-苯乙醇、33%乙醛和10倍乙酸乙酯产量,产生低于二者纯种培养的0.32 g·L-1挥发酸。1.13 酒香酵母属（Brettanomyces）/德克酵母属（Dekkera）
酒香酵母属中与葡萄酒最密切相关的是布鲁塞尔酒香酵母（Brettanomyces bruxellensis）。酒香酵母对营养的需求很低,耐受高乙醇、高酸、高盐和高糖,对葡萄酒的污染主要发生在酿造过程后期或储存期间。产生乙基酚类物质是其导致葡萄酒腐败的主要原因,有氧条件下可高产乙酸[58],造成红葡萄酒的异味。B. bruxellensis乙烯基还原酶活性很高,可以将羟基肉桂酸转化成挥发酚并产生带有焦糖味的四氢吡啶、带有丁香或熏肉味的4-乙基苯酚、4-乙基愈创木酚和带有汗臭味的异戊酸等物质[59]。4-乙基苯酚的产生主要依赖于B. bruxellensis的存在,而与葡萄品种无关[60]。这些物质在合理范围内能增加酒的复杂度,但过量则会带来牛棚、马厩和老鼠骚味。发酵和保存过程要严格防止污染,选择合适发酵剂接种是避免酒精发酵和苹乳发酵中腐败微生物发展的关键[61]。1.14 其他酵母属
洛德酵母属中与葡萄酒最密切相关的是长孢洛德酵母（Lodderomyces elongisporus）。混合发酵可赋予葡萄酒温带酸果、甜果和香料属性[62]。混合发酵爱格丽（Ecolly）能有效地增加β-大马酮、里哪醇等香气物质含量,增加乙酸酯、脂肪酸、C6-C12脂肪酸乙酯和苯乙基类化合物等发酵香气物质含量,感官品评甜果、酸果、花香等属性小幅度提升,但也增加了生青味[63]。土星拟威尔酵母（Williopsis saturnus）和酿酒酵母混合发酵爱人（Airen）葡萄酒提高了乙酸异戊酯含量[64],混合发酵埃米尔（Emir）葡萄酒具有较高浓度的乙酸、乙酸乙酯和乙酸异戊酯[65]。
Kazachstania gamospora发酵丽波拉葡萄酒可增加乙酸乙酯和乙酯含量,发酵长相思（Sauvignon Blanc）和西拉（Syrah）增加了200倍丙酸苯乙酯含量,Kazachstania aerobia和酿酒酵母混合发酵长相思增加了乙酸乙酯含量[66]。
2 非酿酒酵母对葡萄酒风味的影响机制
酵母的各种代谢产物是影响葡萄酒感官质量的重要因素。非酿酒酵母可以通过不同的代谢通路改变酒精、甘油、挥发性香气物质、甘露糖蛋白/多糖、花色苷等物质的含量,从而影响葡萄酒的颜色、香气和口感（统称风味）。非酿酒酵母一个最显著的特征是具有催化挥发性香气化合物从非挥发性前体释放的高酶活性,还可以通过自身代谢途径或释放胞外酶转化酿酒酵母代谢产物来影响葡萄酒风味。2.1 酒精
在初始糖度为22%—24%的典型发酵中,约95%的糖会被转化为乙醇和CO2,1%转化为细胞物质,其余4%转化为其他终端产物,包括甘油、酸、醇、酯等[67]。非酿酒酵母死亡较早会导致使用非酿酒酵母纯种发酵酒度较低,另外,低葡萄糖-乙醇转化率也会显著降低酒精产量,例如生成1度酒精,酿酒酵母需要17—18 g糖,而非酿酒酵母需要20—22 g糖。这或许与酵母的碳流流向和Crabtree效应有关。耐渗透压能力弱的细胞内碳流会分流流向甘油的合成,从而降低乙醇产量。Crabtree效应是指细胞在高浓度葡萄糖和有氧条件下三羧酸循环和氧化磷酸化受到抑制,转而通过糖酵解产生ATP、乙醇或乳酸。酿酒酵母为Crabtree阳性酵母,在高葡萄糖浓度和有氧条件下合成乙醇;季也蒙毕赤酵母为Crabtree阴性酵母,其糖代谢倾向于呼吸代谢和菌体生产,乙醇合成能力很弱[68]。2.2 甘油
甘油是重要的代谢产物,不挥发,具有轻微甜味和粘稠性,可以增强葡萄酒的圆润感和甜度,提升酒体强度。酵母合成甘油的作用一是抵御外界环境渗透压,二是维持细胞内氧化还原平衡[69]。甘油-丙酮酸途径使甘油增加的同时轻微增加丙酮酸浓度,有利于花青素与丙酮酸结合形成稳定色素Vitisin A [70]。非酿酒酵母普遍耐渗透压能力更弱,因此,甘油合成能力更强。2.3 挥发性香气物质
葡萄酒香气分为一类香气（品种香）、二类香气（发酵香）和三类香气（陈酿香）。非酿酒酵母主要对葡萄酒的一类香气和二类香气有重要影响。形成一类香气的化合物主要包括甲氧基吡嗪、萜烯、C13-降异戊二烯和挥发性硫醇等,呈现花香和果香类香气。其中甲氧基吡嗪具有草本香气,是氨基酸代谢产物[71]。萜烯和C13-降异戊二烯都具有果香花香特征,多以非挥发性糖苷结合态存在,只有降解成挥发性游离态时才能增强感官香气。糖苷主要是葡萄糖苷和二糖苷,二糖苷由阿拉伯糖、阿呋喃糖或鼠李糖取代远端葡萄糖生成。α-L-阿拉伯糖苷酶、α-L-鼠李糖苷酶或β-D-脱氧葡萄糖苷酶首先切割糖苷键,释放相应的单萜烯基-β-D-葡萄糖苷,再由β-葡萄糖苷酶作用释放单萜[72]。浆果自带的β-葡萄糖苷酶受pH、葡萄糖和乙醇抑制,而非酿酒酵母普遍具有比酿酒酵母更高的β-葡萄糖苷酶活性且不受抑制。部分非酿酒酵母还具有阿拉伯糖苷酶、鼠李糖苷酶、脱氧葡萄糖苷酶活性,有利于产生更多的萜烯和降异戊二烯,增强香气[73]。硫醇具有热带水果的特殊香气,主要包括4-巯基-4-甲基-2-戊酮（4MMP）、3-巯基己醇（3MH）和乙酰化形成的3-巯基己基乙酸酯（3MHA）。4MMP和3MH与半胱氨酸或谷胱甘肽结合,以无味的非挥发性前体形式存在,在碳硫裂解酶作用下生成硫醇[74]。形成二类级香气的化合物主要包括挥发性脂肪酸、高级醇、酯类、醛类、挥发酚和含硫化合物,是在发酵过程中形成的香气,因此也叫发酵香,主要由酵母完成。酒中90%的挥发脂肪酸是乙酸,0.2—0.7 g·L-1被认为是最佳浓度,接近风味阈值则令人不快。乙酸的减少还通常伴随着芳香族化合物如乙醛、乙酸乙酯、1-丙醇、正丁醇、1-己醇、2,3-丁二醇和甘油的显著增加[75]。高级醇浓度低于300 mg·L-1时有助于增加葡萄酒的香气复杂性,超过400 mg·L-1则对香气有负面影响[76],通常非酿酒酵母的高级醇合成比酿酒酵母要少。2-苯乙醇在低浓度下具有强烈的玫瑰香味,部分非酿酒酵母合成这种物质的能力更强。酯类中最丰富的是乙酸酯类。乙酸乙酯含量小于50 mg·L-1时有怡人的白兰地和苹果香气,150—200 mg·L-1则会有异味,假丝酵母、毕赤酵母等非酿酒酵母比酿酒酵母产生更多的乙酸乙酯且浓度在限值内。有孢汉逊酵母是2-苯乙酸乙酯和乙酸异戊酯的强生产者,毕赤酵母和红酵母也可产生高水平的乙酸异戊酯[77]。醛类具有苹果气味和花香,感官阈值低且90%以上为乙醛,非酿酒酵母产乙醛能力具有属种特异性和菌株特异性。挥发酚主要是白葡萄酒中的乙烯苯酚和红葡萄酒中的乙基苯酚,一般不受欢迎。酒香酵母、季也蒙毕赤酵母高产乙基苯酚,但H. guilliermondii、H. osmophila和P. membranifaciens均不能脱羧阿魏酸生成挥发酚[78]。含硫化合物有不良的臭鸡蛋和煮白菜气味,感官阈值低、化学反应活泼,难以去除,通常会对葡萄酒的质量产生消极影响。2.4 甘露糖蛋白/多糖
多糖可以改善葡萄酒口感和酒体,增强甜度、圆润度、芳香持久性、蛋白质和酒石酸稳定性,通过与单宁相互作用降低涩度,保护酚类化合物免受氧化,促进苹乳发酵,改善起泡酒的泡沫质量,吸附赭曲霉毒素A[79]。非酿酒酵母释放甘露糖蛋白和多糖的能力普遍优于酿酒酵母[56]。DOMIZIO等[80]发现不同单位重量的酵母释放的多糖比重不同,试验中所选用的八个非酿酒酵母表现出比酿酒酵母更高的释放多糖的能力,甘露糖蛋白组成也存在细微差异。另外,多数非酿酒酵母在发酵中死亡较早。酵母死亡会释放甘露糖蛋白和多糖,也会导致多糖释放增加,并对后续发酵和葡萄酒的口感产生影响。非酿酒酵母一般还具有高果胶酶、蛋白酶、脂肪酶活性。果胶酶可增加葡萄汁得率,蛋白酶能促进澄清,脂肪酶能分解葡萄或酵母自溶的脂肪,有助于葡萄酒酿造和增香。245株非酿酒酵母胞外果胶酶、蛋白酶、β-葡聚糖酶、苔聚糖酶、β-葡萄糖苷酶、纤维素酶、木糖酶、淀粉酶和亚硫酸盐还原酶活性表明其产酶数量和种类具有菌株特异性[81]。
综上,非酿酒酵母的作用机制一是高产多种胞外水解酶,最重要的是β-葡萄糖苷酶,促进萜烯和C13-降异戊二烯释放,增强品种香气。二是独特的生理代谢途径会增加甘油、醇、酯等次级代谢产物种类和含量。三是释放更多的甘露糖蛋白/多糖改善口感。
3 酵母之间的竞争作用
发酵初期的高糖、低pH、SO2、重金属离子,发酵中的厌氧环境、不断降低的营养物质和增高的酒精含量、各种毒性化合物和细胞接触等都是发酵环境对酵母菌群的选择压力,会改变菌群动态从而影响发酵走向。促进酵母生长的化学因素包括碳源、氮源、少量氧气、维生素、矿物质、麦角固醇、不饱和脂肪酸、脂质、镁离子等。抑制酵母生长的化学因素包括酒精、SO2、乙酸、脂肪酸、硫、农药残留、嗜杀因子等。影响优势酵母的第一个因素是快速高浓度乙醇的产生。酿酒酵母产生高浓度乙醇抑制其他菌群生长是形成优势地位的主要因素,被认为是由毒性代谢物介导的抑制作用最显著的例证。乙醇会破坏细胞膜结构完整性进而改变通透性造成细胞死亡,且乙醇毒性会随着pH的降低而增加。第二个因素是SO2。但其有效性会受葡萄汁类型和起始菌种数量影响,50—100 mg·L-1的SO2对红葡萄酒中的非酿酒酵母没有影响,但会抑制白葡萄酒中除C. guilliermondii和接合酵母属以外的某些非酿酒酵母[82]。第三个因素是温度。酿酒酵母能耐受较高的温度,可在38℃下继续发酵,而多数非酿酒酵母在超过25℃时已经受到抑制。第四个因素是低氧环境。相比酿酒酵母,非酿酒酵母一般对低氧环境更不耐受,如K. thermotolerans和T. delbrueckii对低氧含量的低耐受性也是导致其竞争力相对较低的部分原因[83]。少量的氧会间接促进发酵,因为氧气的存在导致甾醇和长链不饱和脂肪酸的生物合成,有利于酵母细胞膜的制造及正常功能的行使。第五个因素是对有限营养物的竞争。碳源和氮源作为最重要的营养物质,其含量和可同化程度均影响酵母生长。葡萄醪中可被利用的糖主要是还原糖如葡萄糖和果糖,另外还有少量甘露糖、多糖和糖蛋白经降解后可被利用。果糖偏好型的非酿酒酵母可以避免与酿酒酵母对糖的竞争,增加生存优势。氮源限制会增加发酵时间和非酿酒酵母在后期发酵中的比例。对氮源的竞争主要是对氨基酸的竞争,酵母对氨基酸的利用存在偏好性,顺序接种中的后接种酵母常会由于氮源耗尽而不能良好繁殖。如在I. orientalis和酿酒酵母混合体系中,I. orientalis代谢精氨酸、亮氨酸、半胱氨酸和苏氨酸的速率较酿酒酵母快,导致酿酒酵母对这几种氨基酸的吸收不足,而且I. orientalis生成的组氨酸对酿酒酵母的生长也造成抑制[50]。第六个因素是毒性中链脂肪酸和有机酸的影响。I. orientalis特异性生成 2.52 g·L-1乳酸,酿酒酵母特异性生成1.14 g·L-1乙酸,但混合体系中 I. orientalis可吸收代谢酿酒酵母产生的乙酸,而I. orientalis产生的乳酸则对酿酒酵母的生长有抑制作用[50]。第七个因素是细胞接触。利用交互式光捕获系统发现被酿酒酵母包围生长的H. uvarum增代时间要比自由状态长15%[84]。利用双室发酵罐研究也发现酿酒酵母和T. delbrueckii之间的物理接触/接近导致T. delbrueckii的快速死亡。当与酿酒酵母物理分离共培养时,T. delbrueckii比常规混合培养具有更高的存活力[85]。细胞间的紧密接触也会造成细胞絮凝和水平基因转移。第八个因素是群体感应。这种传感机制基于信号小分子的产生、分泌和检测,其浓度与培养基中分泌微生物的丰度相关。碳酸氢盐、乙醛、氨、法尼醇、色酚和苯乙醇已被鉴定为群体感应分子[86]。T. delbrueckii和酿酒酵母在混合共发酵双隔离生物反应器系统中,群体感应或许导致了酵母形态从丝状到菌丝体的转变[87]。酵母对群体感应活性分子的识别和相应基因表达的改变值得进一步探究。第九个因素是嗜杀因子。嗜杀因子通常是蛋白质或糖蛋白,是酵母在竞争环境下特异性分泌的活性外毒素,可以杀死亲缘关系较近的酵母菌、丝状真菌或细菌,在葡萄酒发酵环境下具有较高的嗜杀活性。嗜杀因子与细胞壁上的受体如葡聚糖、甘露聚糖、几丁质等结合后转位至细胞膜,与细胞膜上受体相互作用,破坏细胞膜完整性,使细胞裂解死亡;也可以与相应受体或酶结合,扰乱细胞分裂周期,使细胞分裂停留或造成细胞死亡[88]。嗜杀性酿酒酵母K1、K2、K3、K28和K3GR1杀菌谱相对较窄,只对特定菌种起作用,包括众多属种在内的嗜杀性非酿酒酵母K4—K11则具有广谱抗菌能力。酿酒酵母在混合发酵过程中产生阳离子多肽（甘油醛磷酸脱氢酶的一个片段）,可以杀死B. bruxellensis、K. thermotolerans等非酿酒酵母[89]。Z. bailii分泌蛋白毒素Zygocin,通过破坏细胞膜功能行使广谱嗜杀性[90]。M. pulcherrima产生铁-二肽复合色素pulcherrimin,通过耗尽培养基中的铁来干扰其他微生物的生长[91]。目前嗜杀因子的产生机制和作用机制尚未完全明晰,在今后一段相当长时间内都将是值得探索的研究重点。
上述提到的种种效应最终会导致酵母之间的正、中性或负相互作用。如SADOUDI等[92]发现M. pulcherrima与酿酒酵母存在协同效应（正相互作用）,混合发酵的芳香族化合物水平均高于纯种发酵;T. delbrueckii和酿酒酵母存在中性相互作用,混合培养的香气分布非常接近纯种发酵;C. zemplinina与酿酒酵母可能存在负相互作用。这些相互作用都与生物量无关。K. thermotolerans与酿酒酵母也存在协同效应,混合发酵比纯种发酵均产生更多的甘油和苯乙醇[53]。
4 小结与展望
一般而言,相比酿酒酵母纯种发酵,非酿酒酵母和酿酒酵母混合发酵可以获得风味更佳且更能反映特定葡萄酒产区独特风格的葡萄酒。目前已商业化的非酿酒酵母有戴尔有孢圆酵母、耐热克鲁维酵母、美极梅奇酵母、核果梅奇酵母、克鲁维毕赤酵母和粟酒裂殖酵母等。目前生产中应用最多的是通过添加非酿酒酵母与酿酒酵母混合发酵以增加萜烯或酯类物质的种类和含量以及增加多糖释放等,从而增加葡萄酒中的果香和花香,并改善葡萄酒的口感。除此之外,粟酒裂殖酵母降解苹果酸和耐热克鲁维酵母高产乳酸的特性也非常值得关注。需要注意的是,非酿酒酵母对发酵和成酒质量的影响取决于葡萄品种、成熟度、酵母属种、菌株、接种时间、接种率和酿造方式（冷浸渍、发酵温度、苹乳发酵、陈酿等）等,过多的影响因素意味着商业化应用需要更多的实践尝试。原生微生物能更好地适应本地葡萄醪化学环境,进一步根据酿造性能和感官品质筛选优良的本土酵母,开发新的工业用酵母产品,有助于我国葡萄酒发展的本土化、特色化。目前从实验室、小试、中试到工厂大规模生产水平的混合发酵试验逐渐增多。但酒庄的具体实践还需在不同的葡萄果实、酵母种类、酿造方式和目标葡萄酒类型之间进行尝试才行。关于非酿酒酵母基因多样性和遗传学研究也在迅速发展,基因组学、转录组学、蛋白组学、代谢组学和高通量技术结合生物信息学分析将有助于更好理解酵母相互作用、代谢机理和种群动态学。相信随着人们对非酿酒酵母的深入认识,非酿酒酵母在葡萄酒酿造中的应用前景广阔。
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文中引用次数倒序
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URLPMID:25388474
DOI:10.1016/j.foodres.2015.06.030URLPMID:28455011 [本文引用: 1]
One of the main opportunities in the use of non-Saccharomyces yeasts is its great intraspecific variability in relation to the synthesis of secondary products of fermentation. Thus, mixed or sequential fermentation with non-Saccharomyces can increase the synthesis of certain metabolites that are important for colour stability, such as acetaldehyde and pyruvic acid (vitisin precursors) or vinylphenols (vinylphenolic pyranoanthocyanin precursors). Furthermore, the selection and use of non-Saccharomyces yeast strains with good yields in the production of certain volatile compounds (ethyl lactate, 2,3-butanediol, 2-phenylethyl acetate), with limited formation of higher alcohols, is a way to improve the aromatic profile of red wine. The main aim of this work was to evaluate the influence of sequential and mixed fermentations with Schizosaccharomyces pombe and Torulaspora delbrueckii strains on red wine's sensory quality. Anthocyanins and aromatic profiles, as well as glycerol and organic acid content, were analysed in the red wines obtained. Results show that, in general, mixed fermentations can promote an increment in polyols synthesis, while sequential fermentations can enhance the herbaceous aroma. Moreover, the use of T. delbrueckii in mixed fermentations allowed an increase to the fruity character of red wine. The use of S. pombe in sequential fermentations increased the stability of the colouring matter by favouring vitisins and vinylphenolic pyranoanthocyanin formation.
DOI:10.1016/j.ijfoodmicro.2015.04.037URLPMID:26001522 [本文引用: 1]
The aim of this work was to study ester formation and the aromatic impact of Torulaspora delbrueckii when used in association with Saccharomyces cerevisiae during the alcoholic fermentation of must. In order to evaluate the influence of the inoculation procedure, sequential and simultaneous mixed cultures were carried out and compared to pure cultures of T. delbrueckii and S. cerevisiae. Our results showed that mixed inoculations allowed the increase, in comparison to S. cerevisiae pure culture, of some esters specifically produced by T. delbrueckii and significantly correlated to the maximal T. delbrueckii population reached in mixed cultures. Thus, ethyl propanoate, ethyl isobutanoate and ethyl dihydrocinnamate were considered as activity markers of T. delbrueckii. On the other hand, isobutyl acetate and isoamyl acetate concentrations were systematically increased during mixed inoculations although not correlated with the development of either species but were rather due to positive interactions between these species. Favoring T. delbrueckii development when performing sequential inoculation enhanced the concentration of esters linked to T. delbrueckii activity. On the contrary, simultaneous inoculation restricted the growth of T. delbrueckii, limiting the production of its activity markers, but involved a very important production of numerous esters due to more important positive interactions between species. These results suggest that the ester concentrations enhancement via interactions during mixed modalities was due to S. cerevisiae production in response to the presence of T. delbrueckii. Finally, sensory analyses showed that mixed inoculations between T. delbrueckii and S. cerevisiae allowed to enhance the complexity and fruity notes of wine in comparison to S. cerevisiae pure culture. Furthermore, the higher levels of ethyl propanoate, ethyl isobutanoate, ethyl dihydrocinnamate and isobutyl acetate in mixed wines were found responsible for the increase of fruitiness and complexity.
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DOI:10.1016/j.ijfoodmicro.2017.06.028URLPMID:28668728 [本文引用: 1]
In last years, non-Saccharomyces yeasts have emerged as innovative tools to improve wine quality, being able to modify the concentration of sensory-impact compounds. Among them, varietal thiols released by yeasts, play a key role in the distinctive aroma of certain white wines. In this context, Torulaspora delbrueckii is in the spotlight because of its positive contribution to several wine quality parameters. This work studies the physiological properties of an industrial T. delbrueckii strain, for the production of wines with increased thiol concentrations. IRC7 gene, previously described in S. cerevisiae, has been identified in T. delbrueckii, establishing the genetics basis of its thiol-releasing capability. Fermentations involving T. delbrueckii showed improvements on several parameters (such as glycerol content, ethanol index, and major volatile compounds composition), but especially on thiols release. These results confirm the potential of T. delbrueckii on wine improvement, describing new metabolic features regarding the release of cysteinylated aroma precursors.
DOI:10.1016/j.fm.2017.07.018URLPMID:28941909 [本文引用: 1]
Today, many non-Saccharomyces strains have been verified can be positive for the development of wine anthocyanin and aroma in different fermentation scenarios. Moreover, oenological tannins are widely used in wine industry to improve the colour profile and aroma complexity. The aim of this work is to analyze the fermentation characters of non-Saccharomyces strains and investigate the effects of pre-fermentative addition of oenological tannins on the wine components as well as sensory properties. For this purpose, five selected non-Saccharomyces strains and grape seed tannin were used to carry out the different fermentation trials. As a result, the grape seed tannin were less likely to influence growth kinetics of non-Saccharomyces strains. Schizosaccharomyces pombe has been proved can be effective to reduce the malic acid content while increase the level of vinylphenolic pyranoanthocyanin, which is positive for wine colour stability. Pre-fermentative use of oenological tannin was verified could be beneficial for the wines fermented with non-Saccharomyces regarding the improvement of wine colour, anthocyanin composition and the complexity of volatile compounds. Nevertheless, sensory analysis showed that oenological tannin could be less effective to modify the aroma impression of non-Saccharomyces wines.
DOI:10.1007/s00217-014-2404-8URL [本文引用: 1]
DOI:10.1016/j.fm.2012.12.011URLPMID:23498200 [本文引用: 1]
Several yeasts were isolated from Campbell Early grapes (Vitis labrusca cultivar Campbell Early), the major grape cultivar in Korea, grown in two different regions. PCR-RFLP analysis of the ITS I-5.8S-ITS II region showed that 34 isolates out of a total of 40 were in the same group. Phylogenetic analysis revealed that the major strain belonged to one species, Hanseniaspora uvarum, although they displayed some nucleotide mismatches between them. During spontaneous alcohol fermentation at 20 degrees C, the two grape musts containing 24 degrees Brix sugar exhibited similar fermentation patterns with differences in final alcohol production and yeast viable counts. PCR analysis of the yeasts randomly isolated during the fermentation using an intron splice site primer showed changes in yeast flora between 8 and 10 days of fermentation. We found that the dominant yeasts displaying various PCR patterns using the primer remained the same throughout the early stages of fermentation, as determined by molecular typing of their ITS regions using PCR-RFLP, and these yeasts were identical to those isolated from grape berries. Among the isolates, the strain designated SS6 was selected based on its potassium metabisulfite resistance, alcohol production (distillation method), and flavor (by sniffing test) of grape juice. When Campbell Early grape must was inoculated with H. uvarum SS6 cells, no differences in fermentation pattern were observed compared with that inoculated with cells of Saccharomyces cerevisiae W-3, an industrial wine yeast strain. However, SS6 wine showed higher levels of organic acid (especially lactic acid), aldehydes, and minor alcohols (except n-propyl alcohol), as well as a higher score in sensory evaluation, compared to those of W-3 wine.
DOI:10.1016/j.foodchem.2017.06.151URLPMID:28873596 [本文引用: 1]
Medium-chain fatty acid (MCFA) ethyl esters, as yeast secondary metabolites, significantly contribute to the fruity aroma of foods and beverages. To improve the MCFA ethyl ester content of wine, mixed fermentations with Hanseniaspora uvarum Yun268 and Saccharomyces cerevisiae were performed. Final volatiles were analyzed by gas solid phase microextraction-chromatography-mass spectrometry, and aroma characteristics were quantitated by sensory analysis. Results showed that mixed fermentation increased MCFA ethyl ester content by 37% in Cabernet Gernischt wine compared to that obtained by pure fermentation. Partial least-squares regression analysis further revealed that the improved MCFA ethyl esters specifically enhanced the temperate fruity aroma of wine. The enhancement of MCFA ethyl esters was attributed to the increased contents of MCFAs that could be induced by the presence of H. uvarum Yun268 in mixed fermentation. Meanwhile, the timing of yeast inoculations significantly affected the involving biomass of each strain and the dynamics of ethanol accumulation.
DOI:10.1007/s00217-016-2757-2URL [本文引用: 1]
DOI:10.3389/fmicb.2016.00670URLPMID:27242698 [本文引用: 1]
In oenology, the utilization of mixed starter cultures composed by Saccharomyces and non-Saccharomyces yeasts is an approach of growing importance for winemakers in order to enhance sensory quality and complexity of the final product without compromising the general quality and safety of the oenological products. In fact, several non-Saccharomyces yeasts are already commercialized as oenological starter cultures to be used in combination with Saccharomyces cerevisiae, while several others are the subject of various studies to evaluate their application. Our aim, in this study was to assess, for the first time, the oenological potential of H. uvarum in mixed cultures (co-inoculation) and sequential inoculation with S. cerevisiae for industrial wine production. Three previously characterized H. uvarum strains were separately used as multi-starter together with an autochthonous S. cerevisiae starter culture in lab-scale micro-vinification trials. On the basis of microbial development, fermentation kinetics and secondary compounds formation, the strain H. uvarum ITEM8795 was further selected and it was co- and sequentially inoculated, jointly with the S. cerevisiae starter, in a pilot scale wine production. The fermentation course and the quality of final product indicated that the co-inoculation was the better performing modality of inoculum. The above results were finally validated by performing an industrial scale vinification The mixed starter was able to successfully dominate the different stages of the fermentation process and the H. uvarum strain ITEM8795 contributed to increasing the wine organoleptic quality and to simultaneously reduce the volatile acidity. At the best of our knowledge, the present report is the first study regarding the utilization of a selected H. uvarum strain in multi-starter inoculation with S. cerevisiae for the industrial production of a wine. In addition, we demonstrated, at an industrial scale, the importance of non-Saccharomyces in the design of tailored starter cultures for typical wines.
DOI:10.1016/j.foodchem.2013.04.056URLPMID:23870989 [本文引用: 1]
Discovery, characterisation and use of novel yeast strains for winemaking is increasingly regarded as a way for improving quality and to provide variation, including subtle characteristic differences in fine wines. The objective of this work was to evaluate the use of a native apiculate strain, selected from grapes, Hanseniaspora vineae (H. vineae) 02/5A. Fermentations were done in triplicate, working with 225 L oak barrels, using a Chardonnay grape must. Three yeast fermentation strategies were compared: conventional inoculation with a commercial Saccharomyces cerevisiae strain, ALG 804, sequential inoculation with H. vineae and then strain ALG 804 and spontaneous fermentation. Yeast strain identification was performed during fermentation, in which the apiculate strain was found to be active, until 9% of alcohol in volume, for the co-fermentation and the spontaneous fermentation was completed by three native S. cerevisiae strains. Basic winemaking parameters and some key chemical analysis, such as concentration of glycerol, biogenic amines, organic acids, and aroma compounds were analysed. Sensory analysis was done using a trained panel and further evaluated with professional winemakers. Sequential inoculation with H. vineae followed by S. cerevisiae resulted in relatively dry wines, with increased aroma and flavour diversity compared with wines resulting from inoculation with S. cerevisiae alone. Wines produced from sequential inoculations were considered, by a winemaker's panel, to have an increased palate length and body. Characteristics of wines derived from sequential inoculation could be explained due to significant increases in glycerol and acetyl and ethyl ester flavour compounds and relative decreases in alcohols and fatty acids. Aroma sensory analysis of wine character and flavour, attributed to winemaking using H. vineae, indicated a significant increase in fruit intensity described as banana, pear, apple, citric fruits and guava. GC analysis of the relative accumulation of 23 compounds to significantly different concentrations for the three fermentation strategies is discussed in relation to aroma compound composition.
DOI:10.3389/fmicb.2016.02160URLPMID:28123382 [本文引用: 1]
In recent decades, the identification of small non-coding RNAs in bacteria has revealed an important regulatory mechanism of gene expression involved in the response to environmental signals and to the control of virulence. In the family Vibrionaceae, which includes several human and animal pathogens, small non-coding RNAs (sRNAs) are closely related to important processes including metabolism, quorum sensing, virulence, and fitness. Studies conducted in silico and experiments using microarrays and high-throughput RNA sequencing have led to the discovery of an unexpected number of sRNAs in Vibrios. The present review discusses the most relevant reports regarding the mechanisms of action of sRNAs and their implications in the virulence of the main human pathogens in the family Vibrionaceae: Vibrio parahaemolyticus, V. vulnificus and V. cholerae.
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DOI:10.1016/j.ijfoodmicro.2009.07.025URLPMID:19683823 [本文引用: 1]
The impact of mixed cultures of Hanseniaspora osmophila and Saccharomyces cerevisiae with different initial yeast ratios on wine composition has been examined. The mixed culture significantly affected sugar consumption, the main enological parameters and ester concentrations, with the exception of glycerol, isoamyl acetate and diethyl succinate levels. Remarkably, in wines obtained with mixed cultures the concentration of 2-phenylethyl acetate was approximately 3- to 9-fold greater than that produced by S. cerevisiae pure culture. Moreover sensory evaluation revealed a stronger fruity character in wines fermented with mixed cultures than in control wines. Independently of the mixed culture used, all wines showed concentrations of acetic acid and ethyl acetate within the ranges described for wines. Our data suggest that a mixed culture of H. osmophila and S. cerevisiae can be used as a tool to increase 2-phenylethyl acetate in wine and that its concentration can be controlled by modulating the initial yeast ratio in the culture.
DOI:10.1111/j.1567-1364.2007.00339.xURLPMID:18179579 [本文引用: 1]
Yeasts identified as Candida stellata are frequently associated with overripe and botrytized grapes and can survive in the fermenting must until the completion of vinification. The molecular taxonomic examination of 41 strains deposited in six culture collections or described in the literature as C. stellata revealed that most of those isolated from grapes or wines belonged to Candida zemplinina and related species. This confusion around the taxonomic position of the strains may account for the rather controversial descriptions of the oenological properties of C. stellata in the literature. Because the authors did not find it among strains newly isolated from botrytized grapes and wines, it was proposed that it is usually C. zemplinina rather than C. stellata that occurs on grapes and in wine fermentation.
DOI:10.1016/j.ijfoodmicro.2015.01.009URLPMID:25625909 [本文引用: 1]
Nowadays, the use of non-Saccharomyces yeasts in combination with Saccharomyces cerevisiae is a state-of-the-art strategy to improve complexity and enhance the analytical composition of the wines. This application has stimulated the interest of understanding how the non-Saccharomyces yeasts can contribute to the quality of the wines. The study presented here explores the potential use of Starmerella bacillaris (synonym Candida zemplinina) under winemaking conditions. Physiological and genetic characterizations of sixty-three isolates of Starm. bacillaris, previously isolated from four different varieties of grapes, were carried out. Both analyses revealed a low level of diversity between the isolates of Starm. bacillaris, while the fermentation trials in laboratory scale demonstrated the good enological performance of this species. The strong fructophilic character of this species and its ability to produce low quantities of ethanol and acetic acid and high amounts of glycerol were confirmed. The results, presented here, demonstrated a potential application of this non-Saccharomyces species in mixed wine fermentations with S. cerevisiae.
DOI:10.1016/j.lwt.2016.06.063URL [本文引用: 1]
URLPMID:22247148 [本文引用: 1]
DOI:10.1016/j.ijfoodmicro.2009.12.025URL [本文引用: 1]
DOI:10.1016/j.micres.2015.02.002URLPMID:25801972 [本文引用: 1]
Zygosaccharomyces bailii BCV 08, a yeast isolated from red wine barrels in Brazil, was evaluated as co-starter in fermentations with Saccharomyces cerevisiae. Z. bailii BCV 08 was preliminarily shown to produce high levels of esters, and the production was optimized in bench and bioreactor scales using grape must. White wine vinifications were conducted with mixed cultures containing different proportions of Z. bailii BCV 08 and an enological strain of S. cerevisiae. In all trials that contained Z. bailii BCV 08, the production of ethyl esters was enhanced in comparison to the vinification control. Our results clearly show the potential of Z. bailii BCV 08 as a mixed starter with S. cerevisiae in order to increase the aromatic complexity of wine.
DOI:10.1016/j.ijfoodmicro.2016.06.004URLPMID:27367967 [本文引用: 1]
Over the last few years the use of multi-starter inocula has become an attractive biotechnological practice in the search for wine with high flavour complexity or distinctive characters. This has been possible through exploiting the particular oenological features of some non-Saccharomyces yeast strains, and the effects that derive from their specific interactions with Saccharomyces. In the present study, we evaluated the selected strain Zygotorulaspora florentina (formerly Zygosaccharomyces florentinus) in mixed culture fermentations with Saccharomyces cerevisiae, from the laboratory scale to the winery scale. The scale-up fermentation and substrate composition (i.e., white or red musts) influenced the analytical composition of the mixed fermentation. At the laboratory scale, mixed fermentation with Z. florentina exhibited an enhancement of polysaccharides and 2-phenylethanol content and a reduction of volatile acidity. At the winery scale, different fermentation characteristics of Z. florentina were observed. Using Sangiovese red grape juice, sequential fermentation trials showed a significantly higher concentration of glycerol and esters while the sensorial analysis of the resulting wines showed higher floral notes and lower perception of astringency. To our knowledge, this is the first time that this yeasts association has been evaluated at the winery scale indicating the potential use of this mixed culture in red grape varieties.
DOI:10.1007/s10295-015-1620-yURLPMID:25903098 [本文引用: 1]
Consumer wine preferences are changing rapidly towards exotic flavours and tastes. In this work, we tested five non-conventional yeast strains for their potential to improve Ribolla Gialla wine quality. These strains were previously selected from numerous yeasts interesting as food production candidates. Sequential fermentation of Ribolla Gialla grape juice with the addition of the Saccharomyces cerevisiae T73 Lalvin industrial strain was performed. Zygosaccharomyces kombuchaensis CBS8849 and Kazachstania gamospora CBS10400 demonstrated positive organoleptic properties and suitable fermentation dynamics, rapid sugar consumption and industrial strain compatibility. At the same time, Torulaspora microellipsoides CBS6641, Dekkera bruxellensis CBS2796 and Dekkera anomala CBS77 were unsuitable for wine production because of poor fermentation dynamics, inefficient sugar consumption and ethanol production levels and major organoleptic defects. Thus, we selected strains of K. gamospora and Z. kombuchaensis that significantly improved the usually plain taste of Ribolla wine by providing additional aromatic complexity in a controlled and reproducible manner.
DOI:10.3864/j.issn.0578-1752.2018.11.015URL [本文引用: 1]
【Objective】It is aimed to evaluate the aroma enhancement of Ecolly dry white wine produced by simultaneous inoculation of one selected Pichia fermentans and Saccharomyces cerevisiae yeast, and to optimize fermentation process of Ecolly dry white wine.【Method】The selected strainP. fermentans (H5Y-28) was co-inoculated in synthetic grape must with a strainS. cerevisiae yeast (F5) at the ratios of 10:1, 4:1, 1:1, 1:4, 1:10, respectively; and single inoculation of S. cerevisiae was set as control. Yeasts biomass and viable cells count of all the samples were monitored during fermentation and the results were used to build yeasts growth kinetics. For fermentation test of Ecolly dry white wine, same ratios of P. fermentans (H5Y-28) and S. cerevisiae yeast (F5)were inoculated on Ecolly must, single inoculation of S. cerevisiae and single inoculation of S. cerevisiae with extracellular enzyme extract from 72h P. fermentans (H5Y-28) culture were used as controls. Volatile aroma compounds analysis and sensory analysis of Ecolly wine samples were conducted by SPME-GC–MS and a well-trained tasting panel in the next April.【Result】Yeasts growth kinetics showed that the amount of viable cells and the survival duration of the selected strain obviously increased with the higher inoculation ratio of P. fermentans in model fermentation, but 10:1 and 4:1 inoculation lowered the total amount of S. cerevisiae yeast. Sensory analysis of Ecolly dry white wine samples revealed that mixed fermentation could improve intensities of fruity and flower characteristics, particularly tropical fruity note. However, higher inoculation ratios of P. fermentans (10:1) caused intense scent of green trait while wine samples of 1:1 and 4:1 only induced weak scent of green trait. Volatile aroma compounds analysis showed that the total concentration of varietal aroma compounds increased significantly as the inoculation ratio of P. fermentans increased, especially terpenols and C13-norisoprenoids. Moreover, the addition of extracellular enzyme extract improved the total level of varietal aroma compounds, especially C13-norisoprenoids (90% higher than the single use of S. cerevisiae). In terms of fermentative aroma compounds, mixed fermentation remarkably elevated the concentration of acetate, C6-C12 fatty acids and their corresponding ethyl esters, isoamyl alcohol and phenyl ethanol. Compared with the wine sample of single use of S. cerevisiae, inoculation ratio of 1:1 improved the contents of varietal aroma compounds and fermentative aroma compounds by 26% and 39%, respectively, and the total content of medium-chain fatty acids ethyl esters in wine sample of 1:1 was notably 40% higher than the control (single use of S. cerevisiae). 【Conclusion】 Mixed fermentation at the ratio 1:1 of P. fermentans and S. cerevisiae has no negative effect on the growth of S. cerevisiae yeast,andhas a great potential to improve the quality of the wine aroma of Ecolly dry white wine.
DOI:10.3864/j.issn.0578-1752.2018.11.015URL [本文引用: 1]
【Objective】It is aimed to evaluate the aroma enhancement of Ecolly dry white wine produced by simultaneous inoculation of one selected Pichia fermentans and Saccharomyces cerevisiae yeast, and to optimize fermentation process of Ecolly dry white wine.【Method】The selected strainP. fermentans (H5Y-28) was co-inoculated in synthetic grape must with a strainS. cerevisiae yeast (F5) at the ratios of 10:1, 4:1, 1:1, 1:4, 1:10, respectively; and single inoculation of S. cerevisiae was set as control. Yeasts biomass and viable cells count of all the samples were monitored during fermentation and the results were used to build yeasts growth kinetics. For fermentation test of Ecolly dry white wine, same ratios of P. fermentans (H5Y-28) and S. cerevisiae yeast (F5)were inoculated on Ecolly must, single inoculation of S. cerevisiae and single inoculation of S. cerevisiae with extracellular enzyme extract from 72h P. fermentans (H5Y-28) culture were used as controls. Volatile aroma compounds analysis and sensory analysis of Ecolly wine samples were conducted by SPME-GC–MS and a well-trained tasting panel in the next April.【Result】Yeasts growth kinetics showed that the amount of viable cells and the survival duration of the selected strain obviously increased with the higher inoculation ratio of P. fermentans in model fermentation, but 10:1 and 4:1 inoculation lowered the total amount of S. cerevisiae yeast. Sensory analysis of Ecolly dry white wine samples revealed that mixed fermentation could improve intensities of fruity and flower characteristics, particularly tropical fruity note. However, higher inoculation ratios of P. fermentans (10:1) caused intense scent of green trait while wine samples of 1:1 and 4:1 only induced weak scent of green trait. Volatile aroma compounds analysis showed that the total concentration of varietal aroma compounds increased significantly as the inoculation ratio of P. fermentans increased, especially terpenols and C13-norisoprenoids. Moreover, the addition of extracellular enzyme extract improved the total level of varietal aroma compounds, especially C13-norisoprenoids (90% higher than the single use of S. cerevisiae). In terms of fermentative aroma compounds, mixed fermentation remarkably elevated the concentration of acetate, C6-C12 fatty acids and their corresponding ethyl esters, isoamyl alcohol and phenyl ethanol. Compared with the wine sample of single use of S. cerevisiae, inoculation ratio of 1:1 improved the contents of varietal aroma compounds and fermentative aroma compounds by 26% and 39%, respectively, and the total content of medium-chain fatty acids ethyl esters in wine sample of 1:1 was notably 40% higher than the control (single use of S. cerevisiae). 【Conclusion】 Mixed fermentation at the ratio 1:1 of P. fermentans and S. cerevisiae has no negative effect on the growth of S. cerevisiae yeast,andhas a great potential to improve the quality of the wine aroma of Ecolly dry white wine.
DOI:10.1016/j.lwt.2017.08.018URL [本文引用: 1]
DOI:10.1007/s00217-014-2220-1URL [本文引用: 1]
DOI:10.1016/j.ijfoodmicro.2016.05.023URLPMID:27261767 [本文引用: 1]
Schizosaccharomyces was initially considered as a spoilage yeast because of the production of undesirable metabolites such as acetic acid, hydrogen sulfide, or acetaldehyde, but it currently seems to be of great value in enology.o ced Nevertheless, Schizosaccharomyces can reduce all of the malic acid in must, leading to malolactic fermentation. Malolactic fermentation is a highly complicated process in enology and leads to a higher concentration of biogenic amines, so the use of Schizosaccharomyces pombe can be an excellent tool for assuring wine safety. Schizosaccharomyces also has much more potential than only reducing the malic acid content, such as increasing the level of pyruvic acid and thus the vinylphenolic pyranoanthocyanin content. Until now, few commercial strains have been available and little research on the selection of appropriate yeast strains with such potential has been conducted. In this study, selected and wild Sc. pombe strains were used along with a Saccharomyces cerevisiae strain to ferment red grape must. The results showed significant differences in several parameters including non-volatile and volatile compounds, anthocyanins, biogenic amines and sensory parameters.
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DOI:10.1007/s00217-017-3015-yURL [本文引用: 1]
DOI:10.1016/j.fm.2016.08.010URLPMID:27697163 [本文引用: 1]
The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3-7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. A higher content of polysaccharide was found in media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe. Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus. A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus. Analysis of the proteins released in the media revealed the presence of a glycoprotein with a molecular size around 32-33 kDa only for the species S. japonicus. Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N-glycan chains released in the media by both yeast species but differences between the two species were also observed. These observations suggest a possible role of rapid MALDI-TOF screening of N-glycans compositional fingerprint as a taxonomic tool for this genus. Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations.
DOI:10.3390/fermentation4010014URL [本文引用: 1]
DOI:10.3390/fermentation4010008URL [本文引用: 1]
DOI:10.3389/fmicb.2016.00610URLPMID:27199939 [本文引用: 1]
Montepulciano d'Abruzzo is a native grape variety of Vitis vinifera L., grown in central Italy and used for production of high quality red wines. Limited studies have been carried out to improve its enological characteristics through the use of indigenous strains of Saccharomyces cerevisiae. The main objective of the present work was to test two indigenous strains of S. cerevisiae (SRS1, RT73), a strain of Starmerella bacillaris (STS12), one of Hanseniaspora uvarum (STS45) and a co-culture of S. cerevisiae (SRS1) and S. bacillaris (STS12), in an experimental cellar to evaluate their role in the sensory characteristic of Montepulciano d'Abruzzo wine. A S. cerevisiae commercial strain was used. Fermentations were conducted under routine Montepulciano d'Abruzzo wine production, in which the main variables were the yeast strains used for fermentation. Basic winemaking parameters, some key chemical analysis and aroma compounds were considered. S. cerevisiae strain dynamics during fermentation were determined by molecular methods. The musts inoculated with the co-culture were characterized by a faster fermentation start and a higher content of glycerol after 3 days of fermentation, as well as the musts added with strains S. bacillaris (STS12) and H. uvarum (STS45). At the end of fermentation the parameters studied were quite similar in all the wines. Total biogenic amines (BA) content of all the wines was low. Ethanolamine was the predominant BA, with a concentration ranging from 21 to 24 mg/l. Wines were characterized by esters and alcohols. In particular, 2-phenylethanol, 3-methylbut-1-yl methanoate, and ethyl ethanoate were the major aroma volatile compounds in all wines. Statistical analysis highlighted the different role played by aroma compounds in the differentiation of wines, even if it was impossible to select a single class of compounds as the most important for a specific yeast. The present study represents a further step toward the use of tailored autochthonous strains to impart the specific characteristics of a given wine which are an expression of a specific terroir.
DOI:10.1007/s00217-015-2497-8URL [本文引用: 1]
DOI:10.1016/j.ijfoodmicro.2017.04.002URLPMID:28436828 [本文引用: 1]
Strategies for production of wines containing lower alcohol concentrations are in strong demand, for reasons of quality, health, and taxation. Development and application of wine yeasts that are less efficient at transforming grape sugars into ethanol has the potential to allow winemakers the freedom to make lower alcohol wines from grapes harvested at optimal ripeness, without the need for post-fermentation processes aimed at removing ethanol. We have recently shown that two non-conventional wine yeast species Metschnikowia pulcherrima and Saccharomyces uvarum were both able to produce wine with reduced alcohol concentration. Both species produced laboratory-scale wines with markedly different volatile aroma compound composition relative to Saccharomyces cerevisiae. This work describes the volatile composition and sensory profiles of reduced-alcohol pilot-scale Merlot wines produced with M. pulcherrima and S. uvarum. Wines fermented with M. pulcherrima contained 1.0% v/v less ethanol than S. cerevisiae fermented wines, while those fermented with S. uvarum showed a 1.7% v/v reduction in ethanol. Compared to S. cerevisiae ferments, wines produced with M. pulcherrima showed higher concentrations of ethyl acetate, total esters, total higher alcohols and total sulfur compounds, while wines fermented with S. uvarum were characterised by the highest total concentration of higher alcohols. Sensorially, M. pulcherrima wines received relatively high scores for sensory descriptors such as red fruit and fruit flavour and overall exhibited a sensory profile similar to that of wine made with S. cerevisiae, whereas the main sensory descriptors associated with wines fermented with S. uvarum were barnyard and meat. This work demonstrates the successful application of M. pulcherrima AWRI3050 for the production of pilot-scale red wines with reduced alcohol concentration and highlights the need for rigorous evaluation of non-conventional yeasts with regard to their sensory impacts.
DOI:10.1007/s10482-015-0578-0URLPMID:26386703 [本文引用: 2]
During certain wine fermentation processes, yeasts, and mainly non-Saccharomyces strains, produce and secrete enzymes such as beta-glucosidases, proteases, pectinases, xylanases and amylases. The effects of enzyme activity on the aromatic quality of wines during grape juice fermentation, using different co-inoculation strategies of non-Saccharomyces and Saccharomyces cerevisiae yeasts, were assessed in the current study. Three strains with appropriate enological performance and high enzymatic activities, BSc562 (S. cerevisiae), BDv566 (Debaryomyces vanrijiae) and BCs403 (Candida sake), were assayed in pure and mixed Saccharomyces/non-Saccharomyces cultures. beta-Glucosidase, pectinase, protease, xylanase and amylase activities were quantified during fermentations. The aromatic profile of pure and mixed cultures was determined at the end of each fermentation. In mixed cultures, non-Saccharomyces species were detected until day 4-5 of the fermentation process, and highest populations were observed in MSD2 (10% S. cerevisiae/90% D. vanrijiae) and MSC1 (1% S. cerevisiae/99% C. sake). According to correlation and multivariate analysis, MSD2 presented the highest concentrations of terpenes and higher alcohols which were associated with pectinase, amylase and xylanase activities. On the other hand, MSC1 high levels of beta-glucosidase, proteolytic and xylanolytic activities were correlated to esters and fatty acids. Our study contributes to a better understanding of the effect of enzymatic activities by yeasts on compound transformations that occur during wine fermentation.
DOI:10.1111/jfds.2002.67.issue-3URL [本文引用: 1]
DOI:10.1111/j.1472-765X.2010.02833.xURLPMID:20337931 [本文引用: 1]
AIMS: To test degradation of malic acid content in wine by immobilized Issatchenkia orientalis KMBL 5774 cells recently isolated from Korean wine pomace as a malic acid-degrading yeast. METHODS AND RESULTS: I. orientalis KMBL 5774 cells were immobilized using a mixture of oriental oak (Quercus variabilis) charcoal with sodium alginate. When the immobilized yeast cells were observed on a scanning electron microscope, cells were efficiently immobilized on the surface area of the charcoal. A Korean wine containing a high level of malic acid was treated with the immobilized yeast cells. The HPLC analysis of the malic acid content in the treated wine showed the malic acid content was reduced to 0.75 mg ml(-1) after treatment from the original content of 8.96 mg ml(-1), representing 91.6% of the malic acid was degraded during the treatment. CONCLUSIONS: The immobilization of the malic acid-degrading yeasts with oriental oak charcoal and sodium alginate is useful for degradation of malic acid in wines containing a high level of malic acid with no significant increase in other acids. SIGNIFICANCE AND IMPACT OF THE STUDY: Malic acid is sometimes detrimental to the quality of wines when present at high concentrations in some varieties. The immobilized I. orientalis KMBL5774 cells appear to be a promising candidate in view of developing biotechnological methods for reduction of malic acid contents in wine.
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The degradation potential of Issatchenkia terricola fermentation to L-malic acid and citric acid was explored. The maximum concentrations of SO2 and alcohol and the minimum pH level that the strain could tolerate were 450 mg/L, 5% and 2, respectively. The medium and fermentation conditions for the degradation of both acids by the strain were optimized based on deacidification rate. When the concentration of the determined optimum nitrogen source tryptone was 0.5 g/100 mL, the deacidification rate reached its maximum level, over 90%. The optimum inoculum size and incubation duration were 1.25 × 106－7.5 × 106 CFU/mL and 60 h, respectively, and the resulting degradation rates of 8－20 g/L L-malic acid and 8－12 g/L critic acid reached over 90%.
URL [本文引用: 1]
The degradation potential of Issatchenkia terricola fermentation to L-malic acid and citric acid was explored. The maximum concentrations of SO2 and alcohol and the minimum pH level that the strain could tolerate were 450 mg/L, 5% and 2, respectively. The medium and fermentation conditions for the degradation of both acids by the strain were optimized based on deacidification rate. When the concentration of the determined optimum nitrogen source tryptone was 0.5 g/100 mL, the deacidification rate reached its maximum level, over 90%. The optimum inoculum size and incubation duration were 1.25 × 106－7.5 × 106 CFU/mL and 60 h, respectively, and the resulting degradation rates of 8－20 g/L L-malic acid and 8－12 g/L critic acid reached over 90%.
DOI:10.1007/s11274-005-8220-3URL [本文引用: 1]
DOI:10.1016/j.fm.2012.10.004URLPMID:23200661 [本文引用: 2]
In the last few years there is an increasing interest on the use of mixed fermentation of Saccharomyces and non-Saccharomyces wine yeasts for inoculation of wine fermentations to enhance the quality and improve complexity of wines. In the present work Lachancea (Kluyveromyces) thermotolerans and Saccharomyces cerevisiae were evaluated in simultaneous and sequential fermentation with the aim to enhance acidity and improve the quality of wine. In this specific pairing of yeast strains in mixed fermentations (S. cerevisiae EC1118 and L. thermotolerans 101), this non-Saccharomyces yeast showed a high level of competitiveness. Nevertheless the S. cerevisiae strain dominated the fermentation over the spontaneous S. cerevisiae strains also under the industrial fermentation conditions. The different condition tested (modalities of inoculum, temperature of fermentation, different grape juice) influenced the specific interactions and the fermentation behaviour of the co-culture of S. cerevisiae and L. thermotolerans. However, some metabolic behaviours such as pH reduction and enhancement of 2-phenylethanol and glycerol, were shown here under all of the conditions tested. The specific chemical profiles of these wines were confirmed by the sensory analysis test, which expressed these results at the tasting level as significant increases in the spicy notes and in terms of total acidity increases.
DOI:10.3390/molecules20069510URLPMID:26016543 [本文引用: 1]
Most red wines commercialized in the market use the malolactic fermentation process in order to ensure stability from a microbiological point of view. In this second fermentation, malic acid is converted into L-lactic acid under controlled setups. However this process is not free from possible collateral effects that on some occasions produce off-flavors, wine quality loss and human health problems. In warm viticulture regions such as the south of Spain, the risk of suffering a deviation during the malolactic fermentation process increases due to the high must pH. This contributes to produce wines with high volatile acidity and biogenic amine values. This manuscript develops a new red wine making methodology that consists of combining the use of two non-Saccharomyces yeast strains as an alternative to the traditional malolactic fermentation. In this method, malic acid is totally consumed by Schizosaccharomyces pombe, thus achieving the microbiological stabilization objective, while Lachancea thermotolerans produces lactic acid in order not to reduce and even increase the acidity of wines produced from low acidity musts. This technique reduces the risks inherent to the malolactic fermentation process when performed in warm regions.The result is more fruity wines that contain less acetic acid and biogenic amines than the traditional controls that have undergone the classical malolactic fermentation.
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DOI:10.1016/j.foodchem.2008.05.098URL [本文引用: 2]
DOI:10.1016/j.ijfoodmicro.2011.03.020URLPMID:21531033 [本文引用: 1]
The use of non-Saccharomyces yeasts that are generally considered as spoilage yeasts, in association with Saccharomyces cerevisiae for grape must fermentation was here evaluated. Analysis of the main oenological characteristics of pure cultures of 55 yeasts belonging to the genera Hanseniaspora, Pichia, Saccharomycodes and Zygosaccharomyces revealed wide biodiversity within each genus. Moreover, many of these non-Saccharomyces strains had interesting oenological properties in terms of fermentation purity, and ethanol and secondary metabolite production. The use of four non-Saccharomyces yeasts (one per genus) in mixed cultures with a commercial S. cerevisiae strain at different S. cerevisiae/non-Saccharomyces inoculum ratios was investigated. This revealed that most of the compounds normally produced at high concentrations by pure cultures of non-Saccharomyces, and which are considered detrimental to wine quality, do not reach threshold taste levels in these mixed fermentations. On the other hand, the analytical profiles of the wines produced by these mixed cultures indicated that depending on the yeast species and the S. cerevisiae/non-Saccharomyces inoculum ratio, these non-Saccharomyces yeasts can be used to increase production of polysaccharides and to modulate the final concentrations of acetic acid and volatile compounds, such as ethyl acetate, phenyl-ethyl acetate, 2-phenyl ethanol, and 2-methyl 1-butanol. (C) 2011 Elsevier B.V.
DOI:10.1002/(ISSN)1097-0010URL [本文引用: 1]
DOI:10.1016/j.foodcont.2017.05.028URL [本文引用: 1]
DOI:10.1007/s00217-015-2559-yURL [本文引用: 1]
DOI:10.1007/s00253-017-8666-xURLPMID:29189899 [本文引用: 1]
Brettanomyces bruxellensis is a common and significant wine spoilage microorganism. B. bruxellensis strains generally detain the molecular basis to produce compounds that are detrimental for the organoleptic quality of the wine, including some classes of volatile phenols that derive from the sequential bioconversion of specific hydroxycinnamic acids such as ferulate and p-coumarate. Although B. bruxellensis can be detected at any stage of the winemaking process, it is typically isolated at the end of the alcoholic fermentation (AF), before the staring of the spontaneous malolactic fermentation (MLF) or during barrel aging. For this reason, the endemic diffusion of B. bruxellensis leads to consistent economic losses in the wine industry. Considering the interest in reducing sulfur dioxide use during winemaking, in recent years, biological alternatives, such as the use of tailored selected yeast and bacterial strains inoculated to promote AF and MLF, are actively sought as biocontrol agents to avoid the
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DOI:10.1111/j.1472-765X.2010.02822.xURLPMID:20214731 [本文引用: 1]
AIM: To examine the growth and survival of Williopsis saturnus strains along with wine yeast Saccharomyces cerevisiae in grape must. METHODS AND RESULTS: For this study, fermentations were performed in sterilized grape must at 18 degrees C. Inoculum level was 5 x 10(6) cells per ml for each yeast. The results showed that W. saturnus yeasts exhibited slight growth and survival depending on the strain, but they died off by day 5. Saccharomyces cerevisiae, however, dominated the fermentation, reaching the population of about 8 log CFU ml(-1). It was observed that ethanol formation was not affected. The concentrations of acetic acid, ethyl acetate and isoamyl acetate were found higher in mixed culture experiments compared to control fermentation. The results also revealed that higher alcohols production was unaffected in general. CONCLUSION: Fermentations did not form undesirable concentrations of flavour compounds, but production of higher levels of acetic acid in mixed culture fermentations may unfavour the usage of W. saturnus in wine making. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information on the behaviour of W. saturnus together with S. cerevisiae during the alcoholic fermentation.
DOI:10.1080/08905436.2012.724038URL [本文引用: 1]
DOI:10.1016/j.lwt.2015.05.018URL [本文引用: 1]
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DOI:10.1080/10408398.2012.658587URLPMID:24915378 [本文引用: 1]
This review discusses the factors that affect the concentrations of methoxypyrazines (MPs) and the techniques used to analyze MPs in grapes, musts, and wines. MPs are commonly studied pyrazines in food science due to their contribution of aroma and flavor to numerous vegetables such as peas and asparagus. They are described as highly odorous compounds with a very low olfactory threshold. The grape varietals that exhibit green or herbaceous aromas that are characteristic of MPs are predominantly Vitis vinifera cv. Cabernet Sauvignon and Sauvignon Blanc, but include others. The most extensively studied MPs include 3-isobutyl-2-methoxypyrazine, 3-isopropyl-2-methoxypyrazine, and 3-sec-butyl-2-methoxypyrazine. It outlines the significance of methoxypyrazines in grapes, musts, and wines in terms of the concentrations that are capable of contributing their sensory characteristics to wines. This review discusses methods for analyzing MPs including gas chromatography-mass spectroscopy (one or two dimension) and high-performance liquid chromatography, the appropriate extraction techniques, and the efficacy of these methods. Additionally, this review explores factors that affect pyrazine content of grapes, must, and wines, such as the effects of different viticultural practices, effects of light exposure and grape maturation, climate, soil, the multi-colored Asian lady beetle and the effects of different vinification processes.
DOI:10.1016/0008-6215(88)80012-0URL [本文引用: 1]
DOI:10.1016/S0740-0020(02)00105-3URL [本文引用: 1]
Abstract
The purpose of this study was to verify whether both wine yeasts belonging to different species (Metschnikowia pulcherrima (2), Debaryomyces hansenii, Kluyveromyces thermotolerans, Saccharomyces cerevisiae (3), Hanseniaspora uvarum, Pichia kluyveri) and a non-wine yeast (Candida molischiana), were able to hydrolyse the terpenoids, norisoprenoids and benzenoids glycosides occurring in Muscat must and transform the produced aglycons. The results obtained confirmed the role played by yeasts in releasing volatile compounds from non-volatile precursors. Among wine yeasts, Hanseniaspora uvarum was able to hydrolyse both glycoconjugated forms of pyranic and furanic oxides of linalool.[本文引用: 1]
DOI:10.1016/j.ijfoodmicro.2004.06.007URLPMID:15698691 [本文引用: 1]
The use of Pichia fermentans in pure cultures and sequential mixtures with Saccharomyces cerevisiae has been studied to improve the aromatic compounds and characteristics of a wine. P. fermentans has proved to be a good starter strains for must fermentation in the winemaking industry. It has shown the same level of sulphur tolerance and the same growth rate as S. cerevisiae. We have demonstrated that only 2 days of must fermentation with P. fermentans in sequential mixtures are enough to increase the following compounds in the wine both qualitatively and quantitatively: acetaldehyde, ethyl acetate, 1-propanol, n-butanol, 1-hexanol, ethyl caprilate, 2,3-butanediol and glycerol. Maintaining this non-Saccharomyces strain in contact with the must for longer periods quantitatively increases the flavour composition.
DOI:10.1007/BF01941764URL [本文引用: 1]
DOI:10.1016/j.fm.2008.04.015URL [本文引用: 1]
Abstract
Thirty-eight yeast strains belonging to the genera Candida, Hanseniaspora, Pichia, Torulaspora and Zygosaccharomyces were screened for ester formation on synthetic microbiological medium. The genera Hanseniaspora and Pichia stood out as the best acetate ester producers. Based on the ester profile Hanseniaspora guilliermondii 11027 and 11102, Hanseniaspora osmophila 1471 and Pichia membranifaciens 10113 and 10550 were selected for further characterization of enological traits. When growing on must H. osmophila 1471, which displayed a glucophilic nature and was able to consume more than 90% of initial must sugars, produced levels of acetic acid, medium chain fatty acids and ethyl acetate, within the ranges described for wine. On the other hand, it was found to be a strong producer of 2-phenylethyl acetate. Our data suggest that H. osmophila 1471 is a good candidate for mixed starters, although the possible interactions with S. cerevisiae deserve further research.[本文引用: 1]
DOI:10.1128/AEM.00302-08URLPMID:18606802 [本文引用: 1]
Stabilization against protein haze was one of the first positive properties attributed to yeast mannoproteins in winemaking. In previous work we demonstrated that deletion of KNR4 leads to increased mannoprotein release in laboratory Saccharomyces cerevisiae strains. We have now constructed strains with KNR4 deleted in two different industrial wine yeast backgrounds. This required replacement of two and three alleles of KNR4 for the EC1118 and T73-4 backgrounds, respectively, and the use of three different selection markers for yeast genetic transformation. The actual effect of the genetic modification was dependent on both the genetic background and the culture conditions. The fermentation performance of T73-4 derivatives was clearly impaired, and these derivatives did not contribute to the protein stability of the wine, even though they showed increased mannoprotein release in vitro. In contrast, the EC1118 derivative with both alleles of KNR4 deleted released increased amounts of mannoproteins both in vitro and during wine fermentation assays, and the resulting wines were consistently less susceptible to protein haze. The fermentation performance of this strain was slightly impaired, but only with must with a very high sugar content. These results pave the way for the development of new commercial strains with the potential to improve several mannoprotein-related quality and technological parameters of wine.
DOI:10.1016/j.fm.2014.04.005URLPMID:24929876 [本文引用: 1]
Eight non-Saccharomyces wine strains, previously selected for their ability to modulate the final concentrations of various volatile compounds and to persist with Saccharomyces cerevisiae in mixed inocula fermentations of grape juice, have been analyzed in the present work to test their ability to release mannoproteins. The eight strains were members of different genera originally isolated from grape: Hansensiaspora osmophila, Lachancea thermotolerans, Metschnikowia pulcherrima, Pichia fermentans, Saccharomycodes ludwigii, Starmerella bacillaris, Torulaspora delbrueckii and Zygosaccharomyces florentinus. A synthetic polysaccharide-free grape juice, was used to characterize the mannoproteins released during the alcoholic fermentation. Mannoproteins profiles were characterized by gel electrophoresis and carbohydrate composition was analyzed both by HPLC and by mass spectrometry. The eight non-Saccharomyces yeasts demonstrated a higher capacity to release polysaccharides compared to S. cerevisiae. The proteins released by the eight yeast strains showed a wide variety of protein sizes, ranging from 25 kDa to greater than 250 kDa. The mass spectrometric profile of the N-glycans ranged from 1600 to 4000 Da and was characteristic for each strain. Detailed investigation of the degree of polymerization of released N-glycans revealed variable composition from 8 to 15 units of monosaccharides.
DOI:10.1046/j.1365-2672.2001.01379.xURLPMID:11442729 [本文引用: 1]
AIMS: The objective of this study was to investigate what types of enzymes are being produced by non-Saccharomyces yeasts isolated from grapes in South Africa vineyards and clarified grape juice. These enzyme profiles could pave the way for attributing specific effects in wine to some of these enzymes produced by so-called wild yeasts associated with grape must. METHODS AND RESULTS: In this study 245 yeast isolates, belonging to the genera Kloeckera, Candida, Debaryomyces, Rhodotorula, Pichia, Zygosaccharomyces, Hanseniaspora and Kluyveromyces were screened for the production of extracellular pectinases, proteases beta-glucanases, lichenases, beta-glucosidases, cellulases, xylanases, amylases and sulphite reductase activity. These yeasts, representing 21 species, were previously isolated from grapes and clarified grape juice. The production of all extracellular hydrolytic enzymes screened for was observed except beta-glucosidase activity. The amount and range of enzymes produced varied with different isolates of the same species. CONCLUSION: This study clearly revealed the potential of non-Saccharomyces wine yeasts to produce a wide range of useful extracellular enzymes during the initial phase of wine fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: Enzymes produced by indigenous yeasts associated with grapes and juice might be harnessed to catalyse desired biotransformations during wine fermentation.
DOI:10.1046/j.1365-2621.2001.t01-1-00462.xURL [本文引用: 1]
DOI:10.1046/j.1365-2672.2001.01426.xURLPMID:11556922 [本文引用: 1]
AIMS: The effect of oxygen on the survival of Torulaspora delbrueckii and Kluyveromyces thermotolerans during mixed culture fermentations in grape juice with Saccharomyces cerevisiae was investigated. METHODS AND RESULTS: Fermentations were carried out in two simple fermentation systems differing in the availability of oxygen. At low available oxygen conditions, T. delbrueckii and K. thermotolerans began to die off after two days of mixed culture fermentation. In filtrates from 2-day-old mixed cultures, single cultures of T. delbrueckii and K. thermotolerans survived and actively produced ethanol to concentrations of approx. 65 and 70 g l-1, respectively, at low available oxygen conditions. Oxygen clearly increased the survival time and decreased the death rate of T. delbrueckii and K. thermotolerans in mixed cultures, whereas it did not affect the growth and survival of S. cerevisiae. CONCLUSION: Our results show that the deaths of T. delbrueckii and K. thermotolerans in mixed cultures at low available oxygen conditions are not due to toxic metabolites produced by the yeasts but rather to the lack of oxygen. Furthermore, they indicate that T. delbrueckii and K. thermotolerans are less tolerant to low available oxygen conditions than S. cerevisiae. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study reveals new knowledge on the mechanisms underlying the succession of yeasts during wine fermentations. This knowledge may be of importance when creating defined, mixed starter cultures for the controlled production of wines with a wide range of flavour compositions.
DOI:10.1016/j.femsle.2005.03.008URLPMID:15796993 [本文引用: 1]
Applying a newly developed user-interactive optical trapping system, we controllably surrounded individual cells of one yeast species, Hanseniaspora uvarum, with viable cells of another yeast species, Saccharomyces cerevisiae, thus creating a confinement of the former. Growth of surrounded and non-surrounded H. uvarum cells was followed under a microscope by determining their generation time. The average generation time of surrounded H. uvarum cells was 15% higher than that of non-surrounded cells, thereby showing that the confinement imposed by viable S. cerevisiae cells on H. uvarum inhibits growth of the latter. This study is the first to demonstrate that confinement is a determinant of growth in a microbial ecosystem.
DOI:10.1007/s00253-012-4660-5URLPMID:23292550 [本文引用: 1]
Alcoholic fermentation of grape must is a complex process, involving several yeast genera and species. The early stages in fermentation are dominated by non-Saccharomyces yeasts that are gradually replaced by the Saccharomyces cerevisiae species, which takes over the fermentation. Quantitative studies have reported the influence of non-Saccharomyces yeast species on wine quality and evaluated their biotechnological interest. The industrial yeast market, which, until recently, exclusively focused on S. cerevisiae, now offers S. cerevisiae/non-Saccharomyces (including Torulaspora delbrueckii) multi-starters. The development of these new mixed industrial starters requires a better understanding of the interaction mechanisms between yeast populations in order to optimize the aromatic impact of the non-Saccharomyces yeast while ensuring complete alcoholic fermentation thanks to S. cerevisiae. For this purpose, a new double-compartment fermentor was designed with the following characteristics: (1) physical separation of two yeast populations, (2) homogeneity of the culture medium in both compartments, (3) fermentation kinetics monitored by weight loss due to CO2 release, and (4) independent monitoring of growth kinetics in the two compartments. This tool was used to compare mixed inoculations of S. cerevisiae/T. delbrueckii with and without physical separation. Our results revealed that physical contact/proximity between S. cerevisiae and T. delbrueckii induced rapid death of T. delbrueckii, a phenomenon previously described and attributed to a cell-cell contact mechanism. In contrast, when physically separated from S. cerevisiae, T. delbrueckii maintained its viability and its metabolic activity had a marked impact on S. cerevisiae growth and viability. The double fermentor is thus a powerful tool for studying yeast interactions. Our findings shed new light on interaction mechanisms described in microorganism populations.
DOI:10.1146/annurev-food-022811-101219URLPMID:23190140 [本文引用: 1]
Microbial interactions are important for the success and safety of food fermentations. Although much indispensable work has described the microbial succession in various fermentations, little is known about how the microbes present interact. Here, we discuss the various mechanisms of microbial interaction from trophic interactions to cell signaling. We also examine the role these interactions play in the dominant microbiota associated with various food fermentations.
DOI:10.1007/s11274-014-1618-zURLPMID:24500666 [本文引用: 1]
Alcoholic fermentation by an oenological strain of Torulaspora delbrueckii in association with an oenological strain of Saccharomyces cerevisiae was studied in mixed and sequential cultures. Experiments were performed in a synthetic grape must medium in a membrane bioreactor, a special tool designed to study indirect interactions between microorganisms. Results showed that the S. cerevisiae strain had a negative impact on the T. delbrueckii strain, leading to a viability decrease as soon as S. cerevisiae was inoculated. Even for high inoculation of T. delbrueckii (more than 20x S. cerevisiae) in mixed cultures, T. delbrueckii growth was inhibited. Substrate competition and cell-to-cell contact mechanism could be eliminated as explanations of the observed interaction, which was probably an inhibition by a metabolite produced by S. cerevisiae. S. cerevisiae should be inoculated 48 h after T. delbrueckii in order to ensure the growth of T. delbrueckii and consequently a decrease of volatile acidity and a higher isoamyl acetate production. In this case, in a medium with a high concentration of assimilable nitrogen (324 mg L(-1)), S. cerevisiae growth was not affected by T. delbrueckii. But in a sequential fermentation in a medium containing 176 mg L(-1) initial assimilable nitrogen, S. cerevisiae was not able to develop because of nitrogen exhaustion by T. delbrueckii growth during the first 48 h, leading to sluggish fermentation.
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DOI:10.1007/s00253-013-5411-yURLPMID:24292082 [本文引用: 1]
Saccharomyces cerevisiae plays a primordial role in alcoholic fermentation and has a vast worldwide application in the production of fuel-ethanol, food and beverages. The dominance of S. cerevisiae over other microbial species during alcoholic fermentations has been traditionally ascribed to its higher ethanol tolerance. However, recent studies suggested that other phenomena, such as microbial interactions mediated by killer-like toxins, might play an important role. Here we show that S. cerevisiae secretes antimicrobial peptides (AMPs) during alcoholic fermentation that are active against a wide variety of wine-related yeasts (e.g. Dekkera bruxellensis) and bacteria (e.g. Oenococcus oeni). Mass spectrometry analyses revealed that these AMPs correspond to fragments of the S. cerevisiae glyceraldehyde 3-phosphate dehydrogenase (GAPDH) protein. The involvement of GAPDH-derived peptides in wine microbial interactions was further sustained by results obtained in mixed cultures performed with S. cerevisiae single mutants deleted in each of the GAPDH codifying genes (TDH1-3) and also with a S. cerevisiae mutant deleted in the YCA1 gene, which codifies the apoptosis-involved enzyme metacaspase. These findings are discussed in the context of wine microbial interactions, biopreservation potential and the role of GAPDH in the defence system of S. cerevisiae.
DOI:10.1016/S1567-1356(02)00126-5URLPMID:12702248 [本文引用: 1]
Abstract
Zygocin, a protein toxin produced and secreted by a killer virus-infected strain of the osmotolerant yeast Zygosaccharomyces bailii, kills a great variety of human and phytopathogenic yeasts and filamentous fungi. Toxicity of the viral toxin is envisaged in a two-step receptor-mediated process in which the toxin interacts with cell surface receptors at the level of the cell wall and the plasma membrane. Zygocin receptors were isolated and partially purified from the yeast cell wall mannoprotein fraction and could be successfully used as biospecific ligand for efficient one-step purification of the 10-kDa protein toxin by receptor-mediated affinity chromatography. Evidence is presented that zygocin-treated yeast cells are rapidly killed by the toxin, and intensive propidium iodide staining of zygocin-treated cells indicated that the toxin is affecting cytoplasmic membrane function, most probably by lethal ion channel formation. The presented findings suggest that zygocin has potential as a novel antimycotic in combating fungal infections.DOI:10.1111/jam.12446URLPMID:24443784 [本文引用: 1]
Aims In the present study, it was investigated the antagonistic behaviour of Metschnikowia pulcherrima, as biocontrol agent, against the main wine yeast species involved in the winemaking process. Methods and Results Seven strains of M.pulcherrima were evaluated for the antimicrobial activity against 114 yeast strains belonging to Pichia, Candida, Hanseniaspora, Kluyveromyces, Saccharomycodes, Torulaspora, Brettanomyces and Saccharomyces genera. Results showed both different inter-generic and intra-generic responses to the antimicrobial action of M.pulcherrima strains. Interestingly, the antimicrobial activity of M.pulcherrima did not have any influence on the growth of Saccharomyces cerevisiae. Instead, M.pulcherrima displayed a broad and effective antimicrobial action on undesired wild spoilage yeasts, such as Brettanomyces/Dekkera, Hanseniaspora and Pichia genera. Fermentation trials carried out in synthetic grape must confirmed the antimicrobial activity of M.pulcherrima, determining the early death of the non-Saccharomyces co-inoculated cultures. Conclusions The antimicrobial activity of M.pulcherrima does not seem due to proteinaceous compounds such as killer phenomenon, but to the pulcherriminic acid (the precursor of pulcherrimin pigment) that depletes iron present in the medium, making it not available to the other yeasts. Significance and Impact of the Study These data agree with and further support the potential use of selected M.pulcherrima strains in controlled multistarter fermentations with S.cerevisiae starter cultures.
URLPMID:22986187 [本文引用: 1]
There has been increasing interest in the use of selected non-Saccharomyces yeasts in co-culture with Saccharomyces cerevisiae. The main reason is that the multistarter fermentation process is thought to simulate indigenous fermentation, thus increasing wine aroma complexity while avoiding the risks linked to natural fermentation. However, multistarter fermentation is characterised by complex and largely unknown interactions between yeasts. Consequently the resulting wine quality is rather unpredictable. In order to better understand the interactions that take place between non-Saccharomyces and Saccharomyces yeasts during alcoholic fermentation, we analysed the volatile profiles of several mono-culture and co-cultures. Candida zemplinina, Torulaspora delbrueckii and Metschnikowia putcherrima were used to conduct fermentations either in mono-culture or in co-culture with S. cerevisiae. Up to 48 volatile compounds belonging to different chemical families were quantified. For the first time, we show that C. zemplinina is a strong producer of terpenes and lactones. We demonstrate by means of multivariate analysis that different interactions exist between the co-cultures studied. We observed a synergistic effect on aromatic compound production when M. pulcherrima was in co-culture with S. cerevisiae. However a negative interaction was observed between C. zemplinina and S. cerevisiae, which resulted in a decrease in terpene and lactone content. These interactions are independent of biomass production. The aromatic profiles of T. delbrueckii and S. cerevisiae in mono-culture and in co-culture are very close, and are biomass-dependent, reflecting a neutral interaction. This study reveals that a whole family of compounds could be altered by such interactions. These results suggest that the entire metabolic pathway is affected by these interactions. (c) 2012 Elsevier Ltd.
