Extracellular expression of keratinase Ker P from Pseudomonas aeruginosa in E. coli

被引：32

作者：

Sharma, Richa ^{[1
]}

Gupta, Rani ^{[1
]}

机构：

[1] Univ Delhi, Dept Microbiol, New Delhi 110021, India

来源：

BIOTECHNOLOGY LETTERS | 2010年 / 32卷 / 12期

关键词：

Fibrin hydrolysis; Haemoglobin hydrolysis; Keratinase; Pseudomonas aeruginosa; PURIFICATION; PROTEIN;

D O I：

10.1007/s10529-010-0361-2

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Keratinase from Pseudomonas aeruginosa KS-1 was expressed constitutively as an extracellular protein in Escherichia coli with high specific activity of 3.7 kU/mg. It was purified fourfold as a 33 kDa monomeric protein by Q-Sepharose ion exchange chromatography with a recovery of 95%. It is a serine protease with optimal activity at pH 9 and 50 degrees C. It was stable from pH 4 to 12 for 1 h with a t(1/2) of 12 min at 70 degrees C. It hydrolyzed haemoglobin > fibrin > feather keratin > azo-casein > casein > meat protein > gelatin. Among synthetic substrates, it efficiently hydrolyzed N-Suc-ala-ala-pro-phe-pNA, N-Suc-ala-ala-ala-pNA, N-Suc-ala-ala-pro-leu-pNA and also plasmin substrate, D-Val-Leu-Lys-pNA

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页码：1863 / 1868

页数：6

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