Mutagenesis and redox partners analysis of the P450 fatty acid decarboxylase OleT(JE)

doi:10.1038/srep44258

QIBEBT-IR > 酶工程研究组

	Mutagenesis and redox partners analysis of the P450 fatty acid decarboxylase OleT(JE)
	Fang, Bo 1,2; Xu, Huifang1 ; Liu, Yi1 ; Qi, Fengxia1 ; Zhang, Wei 1; Chen, Hui 1; Wang, Cong1 ; Wang, Yilin 1,2; Yang, Wenxia1 ; Li, Shengying 1
	2017-03-09
发表期刊	SCIENTIFIC REPORTS
卷号	7
摘要	The cytochrome P450 enzyme OleT(JE) from Jeotgalicoccus sp. ATCC 8456 is capable of converting free long-chain fatty acids into alpha-alkenes via one-step oxidative decarboxylation in presence of H2O2 as cofactor or using redox partner systems. This enzyme has attracted much attention due to its intriguing but unclear catalytic mechanism and potential application in biofuel production. Here, we investigated the functionality of a select group of residues (Arg245, Cys365, His85, and Ile170) in the active site of OleTJE through extensive mutagenesis analysis. The key roles of these residues for catalytic activity and reaction type selectivity were identified. In addition, a range of heterologous redox partners were found to be able to efficiently support the decarboxylation activity of OleTJE. The best combination turned out to be SeFdx-6 (ferredoxin) from Synechococcus elongatus PCC 7942 and CgFdR-2 (ferredoxin reductase) from Corynebacterium glutamicum ATCC 13032, which gave the highest myristic acid conversion rate of 94.4%. Moreover, Michaelis-Menton kinetic parameters of OleTJE towards myristic acid were determined.
文章类型	Article
WOS标题词	Science & Technology
DOI	10.1038/srep44258
关键词[WOS]	CYTOCHROME-P450 ENZYMES ; OXIDATIVE DECARBOXYLATION ; SUBSTRATE RECOGNITION ; BACILLUS-SUBTILIS ; CRYSTAL-STRUCTURE ; HYDROXYLATION ; PEROXYGENASE ; CHLOROPEROXIDASE ; PEROXIDASE ; ALKENES
收录类别	SCI
语种	英语
WOS研究方向	Science & Technology - Other Topics
项目资助者	National Science Foundation of China (NSFC)(31422002 ; Shandong Provincial Natural Science Foundation(JQ201407) ; Applied Basic Research Programs of Science and Technology of Qingdao(15-9-1-106-jch) ; 31270855 ; 21406250)
WOS类目	Multidisciplinary Sciences
WOS记录号	WOS:000395976400001
引用统计
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/9344
专题	酶工程研究组
作者单位	1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biofuels, Shandong Prov Key Lab Synthet Biol, 189 Songling Rd, Qingdao 266101, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Fang, Bo,Xu, Huifang,Liu, Yi,et al. Mutagenesis and redox partners analysis of the P450 fatty acid decarboxylase OleT(JE)[J]. SCIENTIFIC REPORTS,2017,7.
APA	Fang, Bo.,Xu, Huifang.,Liu, Yi.,Qi, Fengxia.,Zhang, Wei.,...&Li, Shengying.(2017).Mutagenesis and redox partners analysis of the P450 fatty acid decarboxylase OleT(JE).SCIENTIFIC REPORTS,7.
MLA	Fang, Bo,et al."Mutagenesis and redox partners analysis of the P450 fatty acid decarboxylase OleT(JE)".SCIENTIFIC REPORTS 7(2017).