S-selective hydroxynitrile lyase from a plant Baliospermum montanum: Molecular characterization of recombinant enzyme

doi:10.1016/j.jbiotec.2011.02.004

	S-selective hydroxynitrile lyase from a plant Baliospermum montanum: Molecular characterization of recombinant enzyme
	Dadashipour, Mohammad 1,2; Yamazaki, Mizue 1,2; Momonoi, Kazumi 3; Tamura, Ken'ichirou 1,2; Fuhshuku, Ken-ichi 1,2; Kanase, Yurina 1,2; Uchimura, Etsuzoh 4; Guan Kaiyun 5; Asano, Yasuhisa 1,2
通讯作者	Asano, Y (reprint author), Toyama Prefectural Univ, Biotechnol Res Ctr, 5180 Kurokawa, Toyama 9390398, Japan
	2011-05-20
发表期刊	JOURNAL OF BIOTECHNOLOGY
ISSN	0168-1656
卷号	153 期号:3-4 页码:100-110
摘要	A novel S-hydroxynitrile lyase (HNL) was purified from leaves of a plant, Baliospermum montanum, by ammonium sulfate fractionation and column chromatographies. Full-length cDNA and genomic DNA were cloned and sequenced. The latter contained two introns and one ORF encoding a 263-residue protein (subunit: 29.5 kDa). The hnl gene was expressed in Escherichia coli and the enzyme was characterized including detailed kinetic studies of 20 substrates for (S)-cyanohydrin synthesis. The enzyme exhibited the highest specific activity (178 U/mg), k(cat) (98/s) and k(cat)/K(m) ratio for piperonal. kcat/Km ratio for aromatic aldehydes was much larger than those of aliphatic aldehydes and ketones. It was strongly inhibited by AgNO(3), PMSF, phenol and methyl ethyl ketone, showed an optimum at pH 5, while having activity at range of 4-6.5. It exhibited stability at wide pH range 2.4-11, the highest activity at 20 degrees C, being active at 0-65 degrees C. The enzyme showed variations in residues involved in substrate pocket and substrate entrance channel compared to other S-selective HNLs, based on a model was built. C-terminal short truncations provided more enzyme production. Gel filtration revealed a 60-65 kDa molecular mass for this non-FAD enzyme and its C-terminally truncated forms using three buffer compositions, indicating dimeric structures. (C) 2011 Elsevier B. V. All rights reserved.
关键词	S-selective Hydroxynitrile Lyase Substrate Specificity Characterization Molecular Cloning Homology Modeling Alanine Scanning
学科领域	Biotechnology & Applied Microbiology
DOI	10.1016/j.jbiotec.2011.02.004
收录类别	SCI
语种	英语
WOS记录号	WOS:000290513700004
引用统计
文献类型	期刊论文
条目标识符	http://ir.kib.ac.cn/handle/151853/5235
专题	资源植物与生物技术所级重点实验室
作者单位	1.Toyama Prefectural Univ, Biotechnol Res Ctr, Toyama 9390398, Japan 2.Toyama Prefectural Univ, Dept Biotechnol, Toyama 9390398, Japan 3.Toyama Agr Res Ctr, Toyama 9398153, Japan 4.Bot Gardens Toyama, Toyama 9392713, Japan 5.Chinese Acad Sci, Kunming Inst Bot, Kunming Bot Garden, Kunming 650204, Yunnan, Peoples R China
推荐引用方式 GB/T 7714	Dadashipour, Mohammad,Yamazaki, Mizue,Momonoi, Kazumi,et al. S-selective hydroxynitrile lyase from a plant Baliospermum montanum: Molecular characterization of recombinant enzyme[J]. JOURNAL OF BIOTECHNOLOGY,2011,153(3-4):100-110.
APA	Dadashipour, Mohammad.,Yamazaki, Mizue.,Momonoi, Kazumi.,Tamura, Ken'ichirou.,Fuhshuku, Ken-ichi.,...&Asano, Yasuhisa.(2011).S-selective hydroxynitrile lyase from a plant Baliospermum montanum: Molecular characterization of recombinant enzyme.JOURNAL OF BIOTECHNOLOGY,153(3-4),100-110.
MLA	Dadashipour, Mohammad,et al."S-selective hydroxynitrile lyase from a plant Baliospermum montanum: Molecular characterization of recombinant enzyme".JOURNAL OF BIOTECHNOLOGY 153.3-4(2011):100-110.

条目包含的文件		下载所有文件
文件名称/大小	文献类型	版本类型	开放类型	使用许可
201204090023.pdf（855KB）			开放获取	--	浏览下载