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A thermophilic cell-free cascade enzymatic reaction for acetoin synthesis from pyruvate
Jia, Xiaojing1,2; Liu, Ying1; Han, Yejun1
2017-06-28
发表期刊SCIENTIFIC REPORTS
ISSN2045-2322
卷号7
摘要

Acetoin (3-hydroxy-2-butanone) is an important bio-based platform chemical with wide applications. In vitro enzyme catalysed synthesis exhibits great feasibility in the production of chemicals with high purity. In the present work, a synthetic pathway involving a two-step continuous reaction was constructed in vitro for acetoin production from pyruvate at improved temperature. Thermostable candidates, acetolactate synthase (coAHASL1 and coAHASL2 from Caldicellulosiruptor owensensis OL) and a-acetolactate decarboxylase (bsALDC from Bacillus subtilis IPE5-4) were cloned, heterologously expressed, and characterized. All the enzymes showed maximum activities at 65-70 degrees C and pH of 6.5. Enzyme kinetics analysis showed that coAHASL1 had a higher activity but lower affinity against pyruvate than that of coAHASL2. In addition, the activities of coAHASL1 and bsALDC were promoted by Mn2+ and NADPH. The cascade enzymatic reaction was optimized by using coAHASL1 and bsALDC based on their kinetic properties. Under optimal conditions, a maximum concentration of 3.36 +/- 0.26 mM acetoin was produced from 10 mM pyruvate after reaction for 24 h at 65 degrees C. The productivity of acetoin was 0.14 mM h(-1), and the yield was 67.80% compared with the theoretical value. The results confirmed the feasibility of synthesis of acetoin from pyruvate with a cell-free enzyme catalysed system at improved temperature.

文章类型Article
WOS标题词Science & Technology
DOI10.1038/s41598-017-04684-8
收录类别SCI
语种英语
关键词[WOS]Alpha-acetolactate Decarboxylase ; Bacterial Acetohydroxyacid Synthase ; Lactis Subsp Lactis ; Mycobacterium-tuberculosis ; Directed Evolution ; Potent Inhibitors ; Cloning ; Purification ; Strategies ; Subunit
WOS研究方向Science & Technology - Other Topics
WOS类目Multidisciplinary Sciences
项目资助者National High Technology Research and Development Programme of China (863 Project)(2014AA021905) ; Open Funding Project of the National Key Laboratory of Biochemical Engineering(Y525041178) ; Chinese Academy of Sciences Strategic Biological Resources Service Network(ZSYS-015) ; 100 Talents Programme of the Chinese Academy of Sciences
WOS记录号WOS:000404268900021
引用统计
文献类型期刊论文
条目标识符http://ir.ipe.ac.cn/handle/122111/22698
专题生化工程国家重点实验室
作者单位1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714
Jia, Xiaojing,Liu, Ying,Han, Yejun. A thermophilic cell-free cascade enzymatic reaction for acetoin synthesis from pyruvate[J]. SCIENTIFIC REPORTS,2017,7.
APA Jia, Xiaojing,Liu, Ying,&Han, Yejun.(2017).A thermophilic cell-free cascade enzymatic reaction for acetoin synthesis from pyruvate.SCIENTIFIC REPORTS,7.
MLA Jia, Xiaojing,et al."A thermophilic cell-free cascade enzymatic reaction for acetoin synthesis from pyruvate".SCIENTIFIC REPORTS 7(2017).
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