High-Level Expression, Purification and Large-Scale Production of L-Methionine gamma-Lyase from Idiomarina as a Novel Anti-Leukemic Drug

CORC > 南海海洋研究所 > 中国科学院南海海洋研究所 > 热带海洋环境国家重点实验室(LTO)

	High-Level Expression, Purification and Large-Scale Production of L-Methionine gamma-Lyase from Idiomarina as a Novel Anti-Leukemic Drug
	Huang, Kui-Ying; Hu, Hai-Yan; Tang, Yan-Lai; Xia, Feng-Geng; Luo, Xue-Qun; Liu, Jian-Zhong
刊名	MARINE DRUGS
	2015
卷号	13 期号:8 页码:5492-5507
关键词	l-Methionine -lyase purification production enzyme leukemia anti-leukemic drug cell proliferation apoptosis
通讯作者	qiushuixian2004@163.com ; huhaiyansysu@163.com ; tangyanlai@126.com ; xiafenggeng@163.com ; L-xuequn@126.com ; lssljz@mail.sysu.edu.cn
中文摘要	l-Methionine -lyase (MGL), a pyridoxal 5-phosphate-dependent enzyme, possesses anti-tumor activity. However, the low activity of MGL blocks the anti-tumor effect. This study describes an efficient production process for the recombinant MGL (rMGL) from Idiomarina constructed using the overexpression plasmid in Escherichia coli BL21 (DE3), purification, and large-scale production. The enzyme produced by the transformants accounted for 53% of the total proteins and accumulated at 1.95 mg/mL using a 500 L fermentor. The enzyme was purified to approximately 99% purity using a high-pressure mechanical homogenizer and nickel (Ni) Sepharose 6 Fast Flow (FF) chromatography. Then, the enzyme was polished by gel filtration, the endotoxins were removed using diethyl-aminoethanol (DEAE) Sepharose FF, and the final product was lyophilized with a vacuum freeze dryer at -35 degrees C. The specific activity of rMGL in the lyophilized powder was up to 108 U/mg. Compared to the control, the enzyme significantly inhibited cellular proliferation in a concentration-dependent manner as tested using the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay and induced cellular apoptosis as analyzed by Annexin V-fluorescein isothiocyanate (FITC) with fluorescence-activated cell sorting (FACS) in leukemia cells. This paper demonstrated the cloning, overexpression, and large-scale production protocols for rMGL, which enabled rMGL to be used as a novel anti-leukemic drug.
学科主题	Pharmacology & Pharmacy
内容类型	期刊论文
源URL	[http://ir.scsio.ac.cn/handle/344004/14934]
专题	南海海洋研究所_热带海洋环境国家重点实验室(LTO)
推荐引用方式 GB/T 7714	Huang, Kui-Ying,Hu, Hai-Yan,Tang, Yan-Lai,et al. High-Level Expression, Purification and Large-Scale Production of L-Methionine gamma-Lyase from Idiomarina as a Novel Anti-Leukemic Drug[J]. MARINE DRUGS,2015,13(8):5492-5507.
APA	Huang, Kui-Ying,Hu, Hai-Yan,Tang, Yan-Lai,Xia, Feng-Geng,Luo, Xue-Qun,&Liu, Jian-Zhong.(2015).High-Level Expression, Purification and Large-Scale Production of L-Methionine gamma-Lyase from Idiomarina as a Novel Anti-Leukemic Drug.MARINE DRUGS,13(8),5492-5507.
MLA	Huang, Kui-Ying,et al."High-Level Expression, Purification and Large-Scale Production of L-Methionine gamma-Lyase from Idiomarina as a Novel Anti-Leukemic Drug".MARINE DRUGS 13.8(2015):5492-5507.