| 高产橙花叔醇的酵母细胞工厂创建 |
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| 引用本文： | 张丽丽,马晓琳,王冬,郁彭,黄璐琦,张学礼,戴住波.高产橙花叔醇的酵母细胞工厂创建[J].中国中药杂志,2017,42(15):2962-2968. |
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| 作者姓名： | 张丽丽 马晓琳 王冬 郁彭 黄璐琦 张学礼 戴住波 |
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| 作者单位： | 天津科技大学 生物工程学院, 天津 300457;中国科学院 天津工业生物技术研究所, 天津 300308;中国科学院系统 微生物工程重点实验室, 天津 300308,天津科技大学 生物工程学院, 天津 300457;中国科学院 天津工业生物技术研究所, 天津 300308;中国科学院系统 微生物工程重点实验室, 天津 300308,中国科学院 天津工业生物技术研究所, 天津 300308;中国科学院系统 微生物工程重点实验室, 天津 300308,天津科技大学 生物工程学院, 天津 300457,中国中医科学院 中药资源中心 道地药材国家重点实验室培育基地, 北京 100700,中国科学院 天津工业生物技术研究所, 天津 300308;中国科学院系统 微生物工程重点实验室, 天津 300308,中国科学院 天津工业生物技术研究所, 天津 300308;中国科学院系统 微生物工程重点实验室, 天津 300308 |
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| 基金项目： | 国家高技术研究发展计划（863）项目（2012AA02A704）；中国科学院青年创新促进会项目（2015138） |
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| 摘 要： | 橙花叔醇为萜类精油成分,具有抗菌、抗肿瘤和抗氧化等多种生物学活性。为了实现在酿酒酵母中异源生产橙花叔醇,该研究首先将猕猴桃Actinidia chinensis来源的橙花叔醇合酶(NES)基因通过密码子优化、人工合成后转入底盘菌株FPP-001中获得工程菌株NES-001,其橙花叔醇产量达到2.71 mg·L~(-1)。在进一步工作中,橙花叔醇合酶的N端通过连接肽GGGS融合法尼烯合酶(FPS)后得到橙花叔醇产量显著提高的工程菌NES-002,其产量是NES-001的59.80倍,达到162.07 mg·L~(-1)。最终,通过恢复NES-002中色氨酸生物合成缺陷基因TRP1得到工程菌NES-003,该菌在高密度发酵体系中橙花叔醇产量能达到1 711.53 mg·L~(-1)。该研究为微生物发酵法生产橙花叔醇等倍半萜化合物提供了基础。
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| 关 键 词： | 精油 橙花叔醇 合成生物学 酿酒酵母 |
| 收稿时间： | 2017/4/13 0:00:00 |
Construction of cell factories for high production of nerolidol in Saccharomyces cerevisiae |
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| ZHANG Li-li,MA Xiao-lin,WANG Dong,YU Peng,HUANG Lu-qi,ZHANG Xue-li and DAI Zhu-bo.Construction of cell factories for high production of nerolidol in Saccharomyces cerevisiae[J].China Journal of Chinese Materia Medica,2017,42(15):2962-2968. |
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| Authors: | ZHANG Li-li MA Xiao-lin WANG Dong YU Peng HUANG Lu-qi ZHANG Xue-li and DAI Zhu-bo |
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| Institution: | College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China;Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China,College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China;Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China,Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China,College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China,State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China,Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China and Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China |
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| Abstract: | Nerolidol is an important constituent of terpenoid essential oil and has excellent anti-tumor, anti-bacterial, and anti-oxidative properties. For realizing heterogenous production of nerolidol, our research firstly integrated the codon-optimized Actinidia sinensis nerolidol synthase gene (NES) into the terpenoid chassis strain FPP-001, and obtained NES-001 that could produce 2.71 mg·L-1 nerolidol. Then, the N-terminal of the NES was fused with FPS by linker peptide GGGS. With this strategy, nerolidol production improved by 59.80-fold, reaching 162.07 mg·L-1. Finally, by introduction of auxotrophic marker TRP1 in NES-002, the resulting strain NES-003 could produce 1 711.53 mg·L-1 by high cell density fermentation method. This study provides the basis for the fermentative production of nerolidol and other sesquiterpenoids. |
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| Keywords: | essential oils nerolidol synthetic biology Saccharomyces cerevisiae |
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