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Mutagenesis and redox partners analysis of the P450 fatty acid decarboxylase OleT(JE)
Fang, Bo1,2; Xu, Huifang1; Liu, Yi1; Qi, Fengxia1; Zhang, Wei1; Chen, Hui1; Wang, Cong1; Wang, Yilin1,2; Yang, Wenxia1; Li, Shengying1
2017-03-09
Source PublicationSCIENTIFIC REPORTS
Volume7
AbstractThe 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.
SubtypeArticle
WOS HeadingsScience & Technology
DOI10.1038/srep44258
WOS KeywordCYTOCHROME-P450 ENZYMES ; OXIDATIVE DECARBOXYLATION ; SUBSTRATE RECOGNITION ; BACILLUS-SUBTILIS ; CRYSTAL-STRUCTURE ; HYDROXYLATION ; PEROXYGENASE ; CHLOROPEROXIDASE ; PEROXIDASE ; ALKENES
Indexed BySCI
Language英语
WOS Research AreaScience & Technology - Other Topics
Funding OrganizationNational 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 SubjectMultidisciplinary Sciences
WOS IDWOS:000395976400001
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Cited Times:18[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qibebt.ac.cn/handle/337004/9344
Collection酶工程研究组
Affiliation1.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
Recommended Citation
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).
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