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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2020, Vol. 40 Issue (3): 163-169    DOI: 10.13523/j.cb.1905041
综述     
老黄酶OYE家族的蛋白质工程的研究进展 *
李炳娟(),刘金锭,廖谊芳,韩文英,刘珂,侯晨露,张磊
天津商业大学生物技术与食品科学学院 天津市食品生物技术重点实验室 天津 300134
Advances in Protein Engineering of the Old Yellow Enzyme OYE Family
LI Bing-juan(),LIU Jin-ding,LIAO Yi-fang,HAN Wen-ying,LIU Ke,HOU Chen-lu,ZHANG Lei
Department of Biotechnology and Food Science, Tianjin University of Commerce,Tianjin Key Laboratory of Food and Biotechnology, Tianjin 300134, China
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摘要：

老黄酶OYE家族酶是一类广泛分布的能够催化烯烃化合物不对称还原的酶类,其能够用于多种手性化合物的制备.分析了OYE家族酶的系统分类及催化反应类型,针对目前该类酶在应用过程中出现的稳定性差,活性低及底物特异性强等问题,综述了蛋白质工程方法对该类酶进行改造的研究进展,为深入研究该家族酶的催化机制及进一步改造提供参考,同时为进一步拓展OYE酶的工业化应用奠定基础.
关键词： OYE家族酶;  蛋白质工程;  理性设计;  不对称还原    
Abstract:

The old yellow enzyme family (OYEs) is a large family which widely distributed and capable of catalyzing the asymmetric reduction of olefin compounds. It can be used in the preparation of various chiral compounds. The system classification and catalytic reaction types of OYEs were analyzed. Meanwhile, protein engineering methods on the modification of the stability, activity and switching the substrate specificity of OYEs are also discussed. Some clues for further study the catalytic mechanism of OYEs and lays the foundation for further expanding the industrial application of OYEs were given.
Key words: Old yellow enzyme family (OYEs)    Protein engineering    Rational design    Asymmetric reduction
收稿日期: 2019-05-23 出版日期: 2020-04-18
ZTFLH:  Q816  
基金资助: * 国家自然科学基金青年基金(31801471,31701172);天津市自然科学基金青年基金(18JCQNJC78900,18JCQNJC09900)
通讯作者: 李炳娟     E-mail: libingjuan2010@163.com
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李炳娟
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引用本文:

李炳娟,刘金锭,廖谊芳,韩文英,刘珂,侯晨露,张磊. 老黄酶OYE家族的蛋白质工程的研究进展 *[J]. 中国生物工程杂志, 2020, 40(3): 163-169.

LI Bing-juan,LIU Jin-ding,LIAO Yi-fang,HAN Wen-ying,LIU Ke,HOU Chen-lu,ZHANG Lei. Advances in Protein Engineering of the Old Yellow Enzyme OYE Family. China Biotechnology, 2020, 40(3): 163-169.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.1905041    或    https://manu60.magtech.com.cn/biotech/CN/Y2020/V40/I3/163

图1  OYE酶催化反应示意图
图2  OYE家族酶的系统分类[4]
  
图3  OYE酶活性的快速检测方法[19]
酶名称 来源 改造目的 改造结果 参考文献
PETN E. nterobacter cloacae PB2 提高酶催化活性 W102→F/I,酶催化速率提高5~6倍 [20]
YqjM B. subtilis C26D/I69T对R-构型产物的转化效率提高了130倍 [21]
NCR Zymomonas mobilis 提高酶的稳定性 第4个loop区截短4个氨基酸,显著改善酶的热稳定性及有机溶剂耐受性 [22]
TsER Thermus thermophilus HB8 改变酶的底物/
产物特异性		 C25G/I67T(R→S)或C25D/I67T(S→R)催化产生的产物发生了产物构型的反转 [23]
OYE1 Saccharomyces carlsbergensis F296S/W116A催化产物构型发生反转(R→S) [24]
MR Pseudomonas putida E134R/L146R突变体对辅酶NADH的亲和力提高了15倍,且与野生型酶相比,突变体也能利用辅酶NADPH作为供氢体 [25]
NCR Zymomonas mobilis 产生新的催化特性 W100I产生了新的催化特性[催化甲基-2-(羟甲基)丙烯酸酯产生S-构型的产物] [23]
表2  OYE家族酶的理性设计
图4  OYE家族酶的序列比对结果
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