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

doi:10.1186/s13068-019-1419-6

	Biochemical characterization of three new α-olefin-producing P450 fatty acid decarboxylases with a halophilic property
	Jiang,Yuanyuan 1,2; Li,Zhong 1,2; Wang,Cong 1; Zhou,Yongjin J.3; Xu,Huifang 1; Li,Shengying 1,4,5
刊名	Biotechnology for Biofuels
	2019-04-08
卷号	12 期号:1
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	AbstractBackgroundThe CYP152 family member OleTJE 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.ResultsWe biochemically characterized three new P450 fatty acid decarboxylases including OleTJH, OleTSQ and OleTSA, with respect to their substrate specificity, steady-state kinetics, and salt effects. These enzymes all act as an OleTJE-like fatty acid decarboxylase being able to decarboxylate a range of straight-chain saturated fatty acids (C8–C20) to various degrees. Site-directed mutagenesis analysis to the lower activity P450 enzyme OleTSA 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 (C8–C12). 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.ConclusionThe P450 fatty acid decarboxylases OleTJE, OleTJH, OleTSQ and OleTSA belong to a novel group of moderate halophilic P450 enzymes. OleTJH from Jeotgalicoccus halophilus shows the decarboxylation activity, kinetic parameters, as well as salt tolerance and stability that are comparable to OleTJE. 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 α-olefin biohydrocarbons.
语种	英语
出版者	BioMed Central
WOS记录号	BMC:10.1186/S13068-019-1419-6
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/165780]
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. 2. 3. 4. 5.
推荐引用方式 GB/T 7714	Jiang,Yuanyuan,Li,Zhong,Wang,Cong,et al. Biochemical characterization of three new α-olefin-producing P450 fatty acid decarboxylases with a halophilic property[J]. Biotechnology for Biofuels,2019,12(1).
APA	Jiang,Yuanyuan,Li,Zhong,Wang,Cong,Zhou,Yongjin J.,Xu,Huifang,&Li,Shengying.(2019).Biochemical characterization of three new α-olefin-producing P450 fatty acid decarboxylases with a halophilic property.Biotechnology for Biofuels,12(1).
MLA	Jiang,Yuanyuan,et al."Biochemical characterization of three new α-olefin-producing P450 fatty acid decarboxylases with a halophilic property".Biotechnology for Biofuels 12.1(2019).