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生物酶法催化瓦伦西亚烯生成圆柚酮 被引量：1

Production of(+)-Nootkatone from(+)-Valencene by Biocatalysis
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摘要 在体外,利用野生型CYP450BM-3对瓦伦西亚烯进行催化,酶-底物复合物催化NADPH氧化的速率为31±1.0 nmol(nmol P450)-1min-1,但催化产物中没有检测到圆柚酮的生成。突变体R47L/Y51F/F87A与底物复合物催化NADPH氧化的速率高于野生型,为79±6.5 nmol(nmol P450)-1min-1,并在催化产物中检测到圆柚酮的生成,但其产物选择性较差,圆柚酮的含量仅占总产物的6.8%。与此同时,检测了另一个突变体A74G/F87V/L188Q对瓦伦西亚烯的催化效果,发现其与底物复合物对NADPH的氧化速率与突变体R47L/Y51F/F87A相当,但产物中圆柚酮的比率更高,达8.0%。 Wild-type CYP450BM.3 showed NADPH oxidation rate of 31 ＋ 1.0 nmol （ nmol P450 ） -＇min-I in vitro, however, （ ＋ ） -nootkatone could not be detected in the products. The R47L/YS1F/F87A mutant had higher activity of 79 ＋ 6.5 nmol （ nmo117450 ） -Imin-l. However, it was much less selective and （ ＋ ） -nootkatone constituted 6.8% of the total products. Meanwhile, another mutant A74G/F87V/L188Q was found to have similar NADPH oxidation activity with the R47IEY51F/F87A mutant but a higher （ ＋ ） -nootkatone ratio of 8.0% was generated.
作者 孟飞 俞春娜 李海峰 谢恬
机构地区 杭州师范大学生物医药与健康研究中心 杭州师范大学生命与环境科学学院
出处 《生物技术通报》 CAS CSCD 北大核心 2012年第8期194-198,共5页 Biotechnology Bulletin
基金 国家自然科学基金项目(81001475) 杭州市科技发展计划项目(20101131N05) 国家重大科技专项子课题(2008ZX07101-006-05-KLNSPC2010A02)
关键词 圆柚酮 瓦伦西亚烯 细胞色素 P450BM-3突变体 生物催化 （ ＋ ） -Nootkatone （ ＋ ） -Valencene Cytoehrome P450SM_3 Mutant Biocatalysis
分类号 TQ655 [化学工程—精细化工]
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参考文献17

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  • 2Girhard M, Machida K, hoh M, et al. Regioselective biooxidation of (+) -valencene by recombinant E. coli expressing CYPIO9B1 from Bacillus subtilis in a two-liquid-phase system. Microbial Cell Factories, 2009, 8 ( 1 ) : 3647.
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生物技术通报
2012年 第8期

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