Purification and characterization of a new L-amino acid oxidase from Daboia russellii siamensis venom

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	Purification and characterization of a new L-amino acid oxidase from Daboia russellii siamensis venom
	Zhong SR 1,2; Jin Y 1; Wu JB 1; Jia YH 3; Xu GL 4; Wang GC 4; Xiong YL 1; Lu QM*1; lvqm@mail.kiz.ac.cn
	2009
发表期刊	TOXICON
ISSN	0041-0101
卷号	54 期号:6 页码:763-771
合作性质	其它
摘要	A new L-amino acid oxidase (designated as DRS-LAAO) was purified from Daboia russellii siamensis venom by ion-exchange, gel filtration and affinity chromatographies. DRS-LAAO is a homodimeric enzyme with a molecular weight of 120.0 kDa as measured by size exclusion chromatography and the monomeric molecular weight of 58.0 kDa as measured by SDS-PAGE under both non-reducing and reducing conditions. The N-terminal amino acid sequence (ADDKNPLEECFREDD) of DRS-LAAO shares high identity with other snake venom L-amino acid oxidases, especially with those isolated from viperid venoms. The enzyme displayed high specificity towards hydrophobic L-amino acids. The best substrate of DRS-LAAO was L-Leu followed by L-Phe and L-Ile, while five substrates - L-Pro, L-Asn, L-Gly, L-Ser and L-Cys were not oxidized. Optimal pH of DRS-LAAO was 8.8. The enzyme showed no hemorrhagic activity even at a dosage of 55.0 mu g. DRS-LAAO dose-dependently inhibited platelet aggregation induced by ADP (83.33 mu M) and TMVA (55.0 nM) with an IC50 value of 32.8 mu g/ml and 32.3 mu g/ml, respectively. The minimum inhibitory concentrations (MICs) of DRS-LAAO against Staphylococci aureus (ATCC 25923), Pseudomonas aeruginosa (ATCC 27853) and Escherichia coli (ATCC 25922) were 9.0, 144.0 and 288.0 mu g/ml, respectively. The minimum bactericidal concentrations (MBCs) of the enzyme for these strains were twice of the MIC values. These results showed that DRS-LAAO had the strongest antimicrobial activity against S. aureus among these three international standard stains. Antibacterial-activities of DRS-LAAO against eight clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates were also tested. The MICs of DRS-LAAO against these isolates ranged from 4.5 to 36.0 mu g/ml. And the MBCs of the enzyme against these isolates ranged from 9.0 to 72.0 mu g/ml.
关键词	Daboia Russellii Siamensis Venom L-amino Acid Oxidase Platelet Aggregation Antimicrobial
资助者	The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest. ; The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest. ; The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest. ; The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest.
收录类别	SCI
语种	英语
资助者	The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest. ; The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest. ; The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest. ; The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee (2007C223M, 2007C102M), Western Light Project of CAS and Chinese National Natural Science Foundation (30770431). The work was financially supported by Natural Science Foundation of Yunnan Science and Technology Committee, No. 2007C223M. And it has no conflict of interest.
文献类型	期刊论文
条目标识符	http://ir.kiz.ac.cn/handle/152453/2825
专题	其他科研部门_天然药物功能蛋白质学科组（赖仞）
通讯作者	lvqm@mail.kiz.ac.cn
作者单位	1.Department of Animal Toxinology, Kunming Institute of Zoology, Chinese Academy of Sciences, 32 Jiaochang East Road, Kunming 650223, China 2.Faculty of Biological & Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China 3.Faculty of Biological & Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China 4.Department of Pharmacy, Kunming General Hospital of Chengdu Military Region, Kunming 650032, China
推荐引用方式 GB/T 7714	Zhong SR,Jin Y,Wu JB,et al. Purification and characterization of a new L-amino acid oxidase from Daboia russellii siamensis venom[J]. TOXICON,2009,54(6):763-771.
APA	Zhong SR.,Jin Y.,Wu JB.,Jia YH.,Xu GL.,...&lvqm@mail.kiz.ac.cn.(2009).Purification and characterization of a new L-amino acid oxidase from Daboia russellii siamensis venom.TOXICON,54(6),763-771.
MLA	Zhong SR,et al."Purification and characterization of a new L-amino acid oxidase from Daboia russellii siamensis venom".TOXICON 54.6(2009):763-771.

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