Isobutanol and 2-ketoisovalerate production by Klebsiella pneumoniae via a native pathway

doi:10.1016/j.ymben.2017.07.003

	Isobutanol and 2-ketoisovalerate production by Klebsiella pneumoniae via a native pathway
	Gu, Jinjie1,2,3 ; Zhou, Jidong 2; Zhang, Zhongxi 2; Kim, Chul Ho 4; Jiang, Biao 2; Shi, Jiping2 ; Hao, Jian 2
	2017-09
发表期刊	METABOLIC ENGINEERING (IF:6.8[JCR-2023],7.6[5-Year])
ISSN	1096-7176
卷号	43 页码:71-84
发表状态	已发表
DOI	10.1016/j.ymben.2017.07.003
摘要	Isobutanol is a valuable chemical and is considered a new generation biofuel. Construction of isobutanol synthesis pathways in bacteria is a hot topic in isobutanol production. Here, we show that an isobutanol synthesis pathway exists naturally in Klebsiella pneumoniae; however, this pathway is dormant in the wild-type bacterium. K. pneumoniae is a 2,3-butanediol producer, and the synthesis pathways of 2,3-butanediol, valine and isobutanol all start from condensation of two pyruvate molecules to yield alpha-acetolactate. Inactivation of alpha-acetolactate decarboxylase (encoded by budA) resulted in alpha-acetolactate flowing into the valine pathway, which led to synthesis of isobutanol and 2-ketoisovalerate (a precursor of isobutanol). ldhA (lactate dehydrogenase) deletion further increased the isobutanol and 2-ketoisovalerate production. In the first step of this pathway, BudB (alpha-acetolactate synthase) was identified as responsible for most of the alpha-acetolactate synthesis. Complementation of ilvBN or ilvIH (isoenzymes of budB) both resulted in a remarkable increase in 2-ketoisovalerate production. Thus, alpha-acetolactate formation is the rate-limiting step of 2-ketoisovalerate production. ilvC (acetohydroxy acid isomeroreductase) and ilvD (dihydroxy acid dehydratase) were identified responsible for 2-ketoisovalerate synthesis from alpha-acetolactate. ipdC, which encodes an indole-3-pyruvate decarboxylase, was identified responsible for most of the isobutyraldehyde formation from 2-ketoisovalerate, and isobutanol production was increased 15.7 fold in the ipdC complementation strain, with a final titer of 2.45 g/L. Isobutanol dehydrogenase activity is distributed across multiple alcohol dehydrogenase enzymes expressed by K. pneumoniae. BudC, DhaT, DhaD and YqhD all had isobutanol dehydrogenase activity in vitro. YqhD uses NADPH as the coenzyme, while the other dehydrogenases use NADH. However, inactivating one or two of these dehydrogenases had no effect on isobutanol production in vivo with isobutyraldehyde as the substrate. These results reveal a novel method for biological production of isobutanol and 2-ketoisovalerate.
关键词	2,3-butanediol Isobutanol 2-ketoisovalerate Klebsiella pneumoniae
收录类别	SCI ; EI
语种	英语
资助项目	KRIBB Research Initiative Program[KGM2211531]
WOS研究方向	Biotechnology & Applied Microbiology
WOS类目	Biotechnology & Applied Microbiology
WOS记录号	WOS:000410479600009
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
EI入藏号	20173504088347
EI主题词	Amino acids ; Bacteria ; Enzymes ; Synthesis (chemical)
EI分类号	Chemical Reactions:802.2 ; Organic Compounds:804.1
WOS关键词	ESCHERICHIA-COLI K-12 ; 2-KETOGLUCONIC ACID PRODUCTION ; GENE REPLACEMENT ; CORYNEBACTERIUM-GLUTAMICUM ; NUCLEOTIDE-SEQUENCE ; 1,3-PROPANEDIOL ; BIOSYNTHESIS ; OPERON ; SYNTHASE ; IDENTIFICATION
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/4643
专题	生命科学与技术学院生命科学与技术学院_特聘教授组_史吉平组生命科学与技术学院_博士生
通讯作者	Shi, Jiping; Hao, Jian
作者单位	1.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China 2.Chinese Acad Sci, Shanghai Adv Res Inst, Lab Biorefinery, 99 Haike Rd, Shanghai 201210, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.KRIBB, Jeonbuk Branch Inst, Biorefinery Res Ctr, Jeongeup 580185, Jeonbuk, South Korea
第一作者单位	生命科学与技术学院
第一作者的第一单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Gu, Jinjie,Zhou, Jidong,Zhang, Zhongxi,et al. Isobutanol and 2-ketoisovalerate production by Klebsiella pneumoniae via a native pathway[J]. METABOLIC ENGINEERING,2017,43:71-84.
APA	Gu, Jinjie.,Zhou, Jidong.,Zhang, Zhongxi.,Kim, Chul Ho.,Jiang, Biao.,...&Hao, Jian.(2017).Isobutanol and 2-ketoisovalerate production by Klebsiella pneumoniae via a native pathway.METABOLIC ENGINEERING,43,71-84.
MLA	Gu, Jinjie,et al."Isobutanol and 2-ketoisovalerate production by Klebsiella pneumoniae via a native pathway".METABOLIC ENGINEERING 43(2017):71-84.