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

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Abstract

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°C. It was stable from pH 4 to 12 for 1 h with a t1/2 of 12 min at 70°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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Acknowledgments

Financial assistance from University of Delhi is duly acknowledged by the authors. R. Sharma acknowledges University Grants Commission for providing her with Senior Research Fellowship.

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  1. Department of Microbiology, University of Delhi, South Campus, New Delhi, 110021, India

    Richa Sharma & Rani Gupta

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  1. Richa Sharma
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  2. Rani Gupta
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Correspondence to Rani Gupta.

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Sharma, R., Gupta, R. Extracellular expression of keratinase Ker P from Pseudomonas aeruginosa in E. coli . Biotechnol Lett 32, 1863–1868 (2010). https://doi.org/10.1007/s10529-010-0361-2

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  • DOI: https://doi.org/10.1007/s10529-010-0361-2

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