A method for efficient expression of Pseudomonas aeruginosa alginate lyase in Pichia pastoris

doi:10.1080/10826068.2014.996233

CORC > 中国农业科学院 > 植物保护研究所

	A method for efficient expression of Pseudomonas aeruginosa alginate lyase in Pichia pastoris
	Yue, Michael M.1; Gong, Wendy W.2; Qiao, Yu 1; Ding, Hongbiao 1
刊名	PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
	2016
卷号	46 期号:2 页码:165-170
关键词	Alginate lyase antibiotic overexpression Pichia pastoris Pseudomonas aeruginosa purification
ISSN号	1082-6068
DOI	10.1080/10826068.2014.996233
通讯作者	Yue, Michael M.
英文摘要	As an eco-friendly biocatalyst for alginate hydrolysis, bacteria-derived alginate lyase (AlgL) has been widely used in research and industries to produce oligosaccharides. However, the cost of AlgL enzyme production remains high due to the low expression and difficulty in purification from bacterial cells. In this study we report an effective method to overexpress the Pseudomonas aeruginosa AlgL (paAlgL) enzyme in Pichia pastoris. Fused with a secretory peptide, the recombinant paAlgL was expressed extracellularly and purified from the culture supernatant through a simple process. The purified recombinant enzyme is highly specific for alginate sodium with a maximal activity of 2,440U/mg. The enzymatic activity remained stable below 45 degrees C and at pH between 4 and 10. The recombinant paAlgL was inhibited by Zn2+, Cu2+, and Fe2+ and promoted by Co2+ and Ca2+. Interestingly, we also found that the recombinant paAlgL significantly enhanced the antimicrobial activity of antibiotics ampicillin and kanamycin against Pseudomonas aeruginosa. Our results introduce a method for efficient AlgL production, the characterization, and a new feature of the recombinant paAlgL as an enhancer of antibiotics against Pseudomonas aeruginosa.
学科主题	Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	TAYLOR & FRANCIS INC
WOS记录号	WOS:000367817000009
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/11545]
专题	植物保护研究所
作者单位	1.Chinese Acad Agr Sci, Feed Res Inst, Beijing 100081, Peoples R China 2.Chinese Acad Agr Sci, Inst Plant Protect, Beijing 100081, Peoples R China
推荐引用方式 GB/T 7714	Yue, Michael M.,Gong, Wendy W.,Qiao, Yu,et al. A method for efficient expression of Pseudomonas aeruginosa alginate lyase in Pichia pastoris[J]. PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY,2016,46(2):165-170.
APA	Yue, Michael M.,Gong, Wendy W.,Qiao, Yu,&Ding, Hongbiao.(2016).A method for efficient expression of Pseudomonas aeruginosa alginate lyase in Pichia pastoris.PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY,46(2),165-170.
MLA	Yue, Michael M.,et al."A method for efficient expression of Pseudomonas aeruginosa alginate lyase in Pichia pastoris".PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY 46.2(2016):165-170.