| Bacillaene酮还原酶结构域的异源表达及底物特异性分析 |
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| 引用本文： | 孙潇慧,车程川,季俊杰,郑舰艇,杨革.Bacillaene酮还原酶结构域的异源表达及底物特异性分析[J].生物工程学报,2015,31(9):1355-1362. |
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| 作者姓名： | 孙潇慧 车程川 季俊杰 郑舰艇 杨革 |
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| 作者单位： | 1 曲阜师范大学 生命科学学院，山东 曲阜 273165；2 中国科学院过程工程研究所 生化工程国家重点实验室，北京 100190,1 曲阜师范大学 生命科学学院，山东 曲阜 273165,2 中国科学院过程工程研究所 生化工程国家重点实验室，北京 100190,2 中国科学院过程工程研究所 生化工程国家重点实验室，北京 100190,1 曲阜师范大学 生命科学学院，山东 曲阜 273165 |
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| 基金项目： | 国家重点基础研究发展计划 (973计划) (No. 2013CB734000)，国家自然科学基金 (Nos. 31370101，31400051)，北京市自然科学基金 (No. 5144031)，青年千人计划和中国科学院启动资金资助。 |
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| 摘 要： | Bacillaene生物合成过程中,聚酮合酶第一个延伸模块的酮还原酶结构域(Bac KR1)既催化α酮基的还原,也催化β酮基的还原,具有天然的底物宽泛性。为进一步研究该结构域的底物特异性,在大肠杆菌中对其进行了异源表达。体外酶学分析表明Bac KR1可以催化聚酮类底物(±)-2-甲基-3-氧代戊酸-乙酰半胱胺硫酯外消旋体的立体选择性还原,仅生成4种非对映异构体中的一种,此外Bac KR1还可以催化环己酮和对氯苯乙酮等非聚酮类底物的还原,暗示了聚酮合酶中酮还原酶结构域作为生物催化剂的潜力。
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| 关 键 词： | 聚酮，酮还原酶，异源表达，立体选择性，底物特异性 |
| 收稿时间： | 2014/11/24 0:00:00 |
Heterologous expression and substrate specificity of ketoreductase domain in bacillaene polyketide synthase |
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| Xiaohui Sun,Chengchuan Che,Junjie Ji,Jianting Zheng and Ge Yang.Heterologous expression and substrate specificity of ketoreductase domain in bacillaene polyketide synthase[J].Chinese Journal of Biotechnology,2015,31(9):1355-1362. |
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| Authors: | Xiaohui Sun Chengchuan Che Junjie Ji Jianting Zheng and Ge Yang |
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| Institution: | 1 Collage of Life Sciences, Qufu Normal University, Qufu 273156, Shandong, China; 2 State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China,1 Collage of Life Sciences, Qufu Normal University, Qufu 273156, Shandong, China,2 State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China,2 State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China and 1 Collage of Life Sciences, Qufu Normal University, Qufu 273156, Shandong, China |
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| Abstract: | The ketoreductase (KR) domain in the first extending module of the polyketide synthase (PKS) catalyzes the reductions of both an a-keto group and a b-keto group in the biosynthesis of bacillaene, suggesting the intrinsic substrate promiscuity. In order to further investigate the substrate specificity, the KR domain (BacKR1) was heterologously overexpressed in Escherichia coli. In vitro enzymatic analysis showed that only one of the four diastereomers was formed in the reduction of the racemic (±)-2-methyl-3-oxopentanoyl-N-acetylcysteamine thioester catalyzed by BacKR1. In addition, BacKR1 was revealed to catalyze the reductions of cyclohexanone and p-chloroacetophenone, indicating the potential of KR domians of PKSs as biocatalysts. |
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| Keywords: | polyketide ketoreductase heterologous expression stereoselectivity substrate specificity |
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