Biochemical characterization of three new alpha-olefin-producing P450 fatty acid decarboxylases with a halophilic property

doi:10.1186/s13068-019-1419-6

	Biochemical characterization of three new alpha-olefin-producing P450 fatty acid decarboxylases with a halophilic property
	Xu, Huifang 2; Jiang, Yuanyuan 2,3; Li, Shengying 2,4,5; Li, Zhong 2,3; Wang, Cong 2; Zhou, Yongjin J.1
刊名	BIOTECHNOLOGY FOR BIOFUELS
	2019-04-08
卷号	12 页码:14
关键词	P450 fatty acid decarboxylase Fatty acid Alkene Biofuel Halophilic enzymes
ISSN号	1754-6834
DOI	10.1186/s13068-019-1419-6
通讯作者	Xu, Huifang(xu_hf@qibebt.ac.cn) ; Li, Shengying(lishengying@sdu.edu.cn)
英文摘要	Background: The CYP152 family member OleT(JE) from Jeotgalicoccus sp. ATCC 8456 has been well-known to catalyze the unusual one-step decarboxylation of free fatty acids towards the formation of terminal alkenes. Efforts to tune up its decarboxylation activity for better production of biological alkenes have been extensively explored via approaches such as site-directed mutagenesis and electron source engineering, but with limited success. To gain more insights into the decarboxylation mechanism and reaction bifurcation (decarboxylation versus hydroxylation), we turned to an alternative approach to explore the natural CYP152 resources for a better variety of enzyme candidates. Results: We biochemically characterized three new P450 fatty acid decarboxylases including OleT(JH), OleT(SQ) and OleT(SA), with respect to their substrate specificity, steady-state kinetics, and salt effects. These enzymes all act as an OleT(JE)-like fatty acid decarboxylase being able to decarboxylate a range of straight-chain saturated fatty acids (C-8-C-20) to various degrees. Site-directed mutagenesis analysis to the lower activity P450 enzyme OleT(SA) revealed a number of key amino acid residues within the substrate-binding pocket (T47F, I177L, V319A and L405I) that are important for delicate substrate positioning of different chain-length fatty acids and thus the decarboxylation versus hydroxylation chemoselectivity, in particular for the mid-chain fatty acids (C-8-C-12). In addition, the three new decarboxylases exhibited optimal catalytic activity and stability at a salt concentration of 0.5 M, and were thus classified as moderate halophilic enzymes. Conclusion: The P450 fatty acid decarboxylases OleT(JE), OleT(JH), OleT(SQ) and OleT(SA) belong to a novel group of moderate halophilic P450 enzymes. OleT(JH) from Jeotgalicoccus halophilus shows the decarboxylation activity, kinetic parameters, as well as salt tolerance and stability that are comparable to OleT(JE). Site-directed mutagenesis of several key amino acid residues near substrate-binding pocket provides important guidance for further engineering of these P450 fatty acid decarboxylases that hold promising application potential for production of alpha-olefin biohydrocarbons.
资助项目	National Science Foundation of China[NSFC 31770844] ; DICPQIBEBT program[DICP & QIBEBT UN201706] ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Chinese Academy of Sciences[QYZDB-SSW-SMC042] ; Shandong Provincial Natural Science Foundation[ZR2017ZB0207]
WOS关键词	DIRECTED EVOLUTION ; CYTOCHROME-P450 PEROXYGENASE ; SP NOV. ; OXIDATIVE DECARBOXYLATION ; MICROBIAL BIOSYNTHESIS ; MALATE-DEHYDROGENASE ; TERMINAL ALKENES ; SUBSTRATE SCOPE ; ENZYMES ; HYDROXYLATION
WOS研究方向	Biotechnology & Applied Microbiology ; Energy & Fuels
语种	英语
出版者	BMC
WOS记录号	WOS:000464115400001
资助机构	National Science Foundation of China ; National Science Foundation of China ; DICPQIBEBT program ; DICPQIBEBT program ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Chinese Academy of Sciences ; Chinese Academy of Sciences ; Shandong Provincial Natural Science Foundation ; Shandong Provincial Natural Science Foundation ; National Science Foundation of China ; National Science Foundation of China ; DICPQIBEBT program ; DICPQIBEBT program ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Chinese Academy of Sciences ; Chinese Academy of Sciences ; Shandong Provincial Natural Science Foundation ; Shandong Provincial Natural Science Foundation ; National Science Foundation of China ; National Science Foundation of China ; DICPQIBEBT program ; DICPQIBEBT program ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Chinese Academy of Sciences ; Chinese Academy of Sciences ; Shandong Provincial Natural Science Foundation ; Shandong Provincial Natural Science Foundation ; National Science Foundation of China ; National Science Foundation of China ; DICPQIBEBT program ; DICPQIBEBT program ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Dalian National Laboratory for Clean Energy (DNL), CAS ; Chinese Academy of Sciences ; Chinese Academy of Sciences ; Shandong Provincial Natural Science Foundation ; Shandong Provincial Natural Science Foundation
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/165618]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Xu, Huifang; Li, Shengying
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Div Biotechnol, Dalian 116023, Peoples R China 2.Chinese Acad Sci, Shandong Prov Key Lab Synthet Biol, CAS Key Lab Biofuels, Qingdao Inst Bioenergy & Bioproc Technol, 189 Songling Rd, Qingdao 266101, Shandong, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.Shandong Univ, State Key Lab Microbial Technol, Qingdao 266237, Shandong, Peoples R China 5.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266237, Shandong, Peoples R China
推荐引用方式 GB/T 7714	Xu, Huifang,Jiang, Yuanyuan,Li, Shengying,et al. Biochemical characterization of three new alpha-olefin-producing P450 fatty acid decarboxylases with a halophilic property[J]. BIOTECHNOLOGY FOR BIOFUELS,2019,12:14.
APA	Xu, Huifang,Jiang, Yuanyuan,Li, Shengying,Li, Zhong,Wang, Cong,&Zhou, Yongjin J..(2019).Biochemical characterization of three new alpha-olefin-producing P450 fatty acid decarboxylases with a halophilic property.BIOTECHNOLOGY FOR BIOFUELS,12,14.
MLA	Xu, Huifang,et al."Biochemical characterization of three new alpha-olefin-producing P450 fatty acid decarboxylases with a halophilic property".BIOTECHNOLOGY FOR BIOFUELS 12(2019):14.