CLC number: Q556
On-line Access: 2019-11-21
Received: 2019-07-08
Revision Accepted: 2019-09-05
Crosschecked: 2019-10-08
Cited: 0
Clicked: 3052
Wen-Na Bao, Zi-Sheng Luo, Shi-Wang Liu, Yuan-Feng Wu, Pei-Lian Wei, Gong-Nian Xiao, Yong Liu. Cloning and characterization of an oxiranedicarboxylate hydrolase from Labrys sp. WH-1[J]. Journal of Zhejiang University Science B, 2019, 20(12): 995-1002.
@article{title="Cloning and characterization of an oxiranedicarboxylate hydrolase from Labrys sp. WH-1",
author="Wen-Na Bao, Zi-Sheng Luo, Shi-Wang Liu, Yuan-Feng Wu, Pei-Lian Wei, Gong-Nian Xiao, Yong Liu",
journal="Journal of Zhejiang University Science B",
volume="20",
number="12",
pages="995-1002",
year="2019",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1900392"
}
%0 Journal Article
%T Cloning and characterization of an oxiranedicarboxylate hydrolase from Labrys sp. WH-1
%A Wen-Na Bao
%A Zi-Sheng Luo
%A Shi-Wang Liu
%A Yuan-Feng Wu
%A Pei-Lian Wei
%A Gong-Nian Xiao
%A Yong Liu
%J Journal of Zhejiang University SCIENCE B
%V 20
%N 12
%P 995-1002
%@ 1673-1581
%D 2019
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1900392
TY - JOUR
T1 - Cloning and characterization of an oxiranedicarboxylate hydrolase from Labrys sp. WH-1
A1 - Wen-Na Bao
A1 - Zi-Sheng Luo
A1 - Shi-Wang Liu
A1 - Yuan-Feng Wu
A1 - Pei-Lian Wei
A1 - Gong-Nian Xiao
A1 - Yong Liu
J0 - Journal of Zhejiang University Science B
VL - 20
IS - 12
SP - 995
EP - 1002
%@ 1673-1581
Y1 - 2019
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1900392
Abstract: Objective: This study aimed to clone and characterize the oxiranedicarboxylate hydrolase (ORCH) from Labrys sp. WH-1. Methods: Purification by column chromatography, characterization of enzymatic properties, gene cloning by protein terminal sequencing and polymerase chain reaction (PCR), and sequence analysis by secondary structure prediction and multiple sequence alignment were performed. Results: The ORCH from Labrys sp. WH-1 was purified 26-fold with a yield of 12.7%. It is a monomer with an isoelectric point (pI) of 8.57 and molecular mass of 30.2 kDa. It was stable up to 55 °C with temperature at which the activity of the enzyme decreased by 50% in 15 min (T5015) of 61 °C and the half-life at 50 °C (t1/2, 50 °C) of 51 min and was also stable from pH 4 to 10, with maximum activity at 55 °C and pH 8.5. It is a metal-independent enzyme and strongly inhibited by Cu2+, Ag+, and anionic surfactants. Its kinetic parameters (Km, kcat, and kcat/Km) were 18.7 mmol/L, 222.3 s−1, and 11.9 mmol/(L·s), respectively. The ORCH gene, which contained an open reading frame (ORF) of 825 bp encoding 274 amino acid residues, was overexpressed in Escherichia coli and the enzyme activity was 33 times higher than that of the wild strain. Conclusions: The catalytic efficiency and thermal stability of the ORCH from Labrys sp. WH-1 were the best among the reported ORCHs, and it provides an alternative catalyst for preparation of
[1]Bao WN, Pan HF, Zhang ZH, et al., 2014. Analysis of essential amino acid residues for catalytic activity of cis-epoxysuccinate hydrolase from Bordetella sp. BK-52. Appl Microbiol Biotechnol, 98(4):1641-1649.
[2]Bao WN, Pan HF, Zhang ZH, et al., 2015. Isolation of the stable strain Labrys sp. BK-8 for
[3]Cheng YQ, Wang L, Pan HF, et al., 2014a. Purification and characterization of a novel cis-epoxysuccinate hydrolase from Klebsiella sp. that produces
[4]Cheng YQ, Pan HF, Bao WN, et al., 2014b. Cloning, homology modeling, and reaction mechanism analysis of a novel cis-epoxysuccinate hydrolase from Klebsiella sp. Biotechnol Lett, 36(12):2537-2544.
[5]Dong S, Liu X, Cui G, et al., 2018. Structural insight into the catalytic mechanism of a cis-epoxysuccinate hydrolase producing enantiomerically pure
[6]Liu ZQ, Li Y, Ping LF, et al., 2007a. Isolation and identification of a novel Rhodococcus sp. ML-0004 producing epoxide hydrolase and optimization of enzyme production. Process Biochem, 42(5):889-894.
[7]Liu ZQ, Li Y, Xu YY, et al., 2007b. Cloning, sequencing, and expression of a novel epoxide hydrolase gene from Rhodococcus opacus in Escherichia coli and characterization of enzyme. Appl Microbiol Biotechnol, 74(1):99-106.
[8]Martin WR, Foster JW, 1955. Production of trans-
[9]Pan HF, Xie ZP, Bao WN, et al., 2008. Isolation and identification of a novel cis-epoxysuccinate hydrolase-producing Bordetella sp. BK-52 and optimization of enzyme production. Acta Microbiol Sin, 48(8):1075-1081 (in Chinese).
[10]https://doi.org/10.13343/j.cnki.wsxb.2008.08.012
[11]Pan HF, Bao WN, Xie ZP, et al., 2010. Molecular cloning and characterization of a cis-epoxysuccinate hydrolase from Bordetella sp. BK-52. J Microbiol Biotechnol, 20(4):659-665.
[12]Pan HF, Xie ZP, Bao WN, et al., 2011. Site-directed mutagenesis of epoxide hydrolase to probe catalytic amino acid residues and reaction mechanism. FEBS Lett, 585(15):2545-2550.
[13]Qiao P, Wu MB, Zhu L, et al., 2015. Enhancing the thermal tolerance of a cis-epoxysuccinate hydrolase via combining directed evolution with various semi-rational redesign methods. J Mol Catal B Enzym, 121:96-103.
[14]Salmanowicz BP, Langner M, Franaszek S, 2014. Charge-based characterisation of high-molecular-weight glutenin subunits from common wheat by capillary isoelectric focusing. Talanta, 129:9-14.
[15]Wang ZQ, Wang YS, Su ZG, 2013. Purification and characterization of a cis-epoxysuccinic acid hydrolase from Nocardia tartaricans CAS-52, and expression in Escherichia coli. Appl Microbiol Biotechnol, 97(6):2433-2441.
[16]Yuan L, Sadiq FA, Liu TJ, et al., 2018. Spoilage potential of psychrotrophic bacteria isolated from raw milk and the thermo-stability of their enzymes. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 19(8):630-642.
[17]Zhu J, Zhu LWS, Yang JH, et al., 2018. Proteomic analysis of human umbilical vein endothelial cells exposed to PM2.5. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 19(6):458-470.
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