Cui, HL; Ma, HT; Cui, YL
; Zhu, XL; Qin, S; Lil, RZ发表期刊JOURNAL OF BIOSCIENCE AND BIOENGINEERING
ISSN1389-1723
2019-12
卷号128期号:6页码:755-765
关键词Cytochrome P450Carotenoids hydroxylasePhaeodactylum tricornutumCloningIdentificationFunctional characterization
研究领域Biotechnology & Applied Microbiology; Food Science & Technology
DOI10.1016/j.jbiosc.2019.06.008
产权排序[Cui, Hongli; Ma, Haotian; Zhu, Xiaoli; Lil, Runzhi] Shanxi Agr Univ, Inst Mol Agr & Bioenergy, Taigu 030801, Peoples R China; [Cui, Yulin; Qin, Song] Chinese Acad Sci, Key Lab Coastal Biol & Biol Resources Utilizat, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China
作者部门海岸带生物学与生物资源保护实验室
英文摘要The diatom microalgal Phaeodactylum tricornutum accumulates a large amount of fucoxanthin. Carotenoids hydroxylases (CHYs) play key roles in fucoxanthin biosynthesis in diatoms. However, not any type of CHYs had been identified in P. tricornutum. In this study, two genes (designated Ptrcyp97b1 and Ptrcyp97b2) were cloned, identified and functionally characterized. They shared high sequence identity (50-94 %) with lutein deficient 1-like proteins from other eukaryotes. The typical catalytic active motifs of cytochrome P450s (CYP) were detected in the amino acid sequences of PtrCYP97B1 and PtrCYP97B2. The two genes were probably due to gene duplication. Ptrcyp97b1 and Ptrcyp97b2 transcriptional expression was up-regulated with distinct patterns under high light conditions. The metabolic profiles of the major carotenoids (beta-carotene, zeaxanthin, diadinoxanthin, diatoxanthin and fucoxanthin) were determined based on the high performance liquid chromatography method. The fucoxanthin and diatoxanthin contents were increased, while the beta-carotene content was decreased. By truncation of the N-terminal trans-membrane anchor or chloroplast transit peptide and addition of a 6 x His-tag, PtrCYP97B1 and PtrCYP97B2 were separately heterologously produced in Escherichia coli and purified by Ni-NTA affinity chromatography. Functional analysis showed that PrtCYP97B2 was able to catalyze the hydroxylation of the beta-rings of beta-carotene to produce zeaxanthin in beta-carotene-accumulating E. coli BL21(DE3) cells. PtrCYP97B1 might have the ability to catalyze the hydroxylation of other substrates other than beta-carotene. These results contribute to the further elucidation of xanthophyll biosynthesis in diatoms. (C) 2019, The Society for Biotechnology, Japan. All rights reserved.
文章类型Article
资助机构China Postdoctoral Science FoundationChina Postdoctoral Science Foundation [2018M631768]; Science and Technology Planning Project of Liaoning Province [20170540047]; Applying Basic Research Planning Project of Shanxi Province (Science and Technology Foundation for Youths) [201801D221250]; Key Research and Development (R&D) Planning Project of Shanxi Province [201803D31063]; State Ministry of Agriculture "948" Project [2014-Z39]; Shanxi Province Key Projects of Coal-based Science and Technology [FT-2014-01]; Shanxi Scholarship Council of China [2015-064]; Key Project of the Key Research and Development Program of Shanxi Province, China [201603D312005]
收录类别SCI
语种英语
关键词[WOS]EXPRESSION PROFILE; BETA-CAROTENE; GENE CLONING; BLUE-LIGHT; FUCOXANTHIN; DIADINOXANTHIN; EVOLUTION; PHOTOSYNTHESIS; BIOSYNTHESIS; DIATOXANTHIN
研究领域[WOS]Biotechnology & Applied Microbiology; Food Science & Technology
WOS记录号WOS:000504527000016
引用统计被引频次：5[WOS][WOS记录][WOS相关记录]
文献类型期刊论文
条目标识符http://ir.yic.ac.cnhttp://ir.yic.ac.cn/handle/133337/24800
专题海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室
作者单位1.Shanxi Agr Univ, Inst Mol Agr & Bioenergy, Taigu 030801, Peoples R China;
2.Chinese Acad Sci, Key Lab Coastal Biol & Biol Resources Utilizat, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China
推荐引用方式
GB/T 7714Cui, HL,Ma, HT,Cui, YL,et al. Cloning, identification and functional characterization of two cytochrome P450 carotenoids hydroxylases from the diatom Phaeodactylum tricornutum[J]. JOURNAL OF BIOSCIENCE AND BIOENGINEERING,2019,128(6):755-765.
APACui, HL,Ma, HT,Cui, YL,Zhu, XL,Qin, S,&Lil, RZ.(2019).Cloning, identification and functional characterization of two cytochrome P450 carotenoids hydroxylases from the diatom Phaeodactylum tricornutum.JOURNAL OF BIOSCIENCE AND BIOENGINEERING,128(6),755-765.
MLACui, HL,et al."Cloning, identification and functional characterization of two cytochrome P450 carotenoids hydroxylases from the diatom Phaeodactylum tricornutum".JOURNAL OF BIOSCIENCE AND BIOENGINEERING 128.6(2019):755-765.
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