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Co-expression of the lipase and foldase of Pseudomonas aeruginosa to a functional lipase in Escherichia coli

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Abstract

The lipA gene, a structural gene encoding for protein of molecular mass 48 kDa, and lipB gene, encoding for a lipase-specific chaperone with molecular mass of 35 kDa, of Pseudomonas aeruginosa B2264 were co-expressed in heterologous host Escherichia coli BL21 (DE3) to obtain in vivo expression of functional lipase. The recombinant lipase was expressed with histidine tag at its N terminus and was purified to homogeneity using nickel affinity chromatography. The amino acid sequence of LipA and LipB of P. aeruginosa B2264 was 99–100% identical with the corresponding sequence of LipA and LipB of P. aeruginosa LST-03 and P. aeruginosa PA01, but it has less identity with Pseudomonas cepacia (Burkholderia cepacia) as it showed only 37.6% and 23.3% identity with the B. cepacia LipA and LipB sequence, respectively. The molecular mass of the recombinant lipase was found to be 48 kDa. The recombinant lipase exhibited optimal activity at pH 8.0 and 37°C, though it was active between pH 5.0 and pH 9.0 and up to 45°C. K m and V max values for recombinant P. aeruginosa lipase were found to be 151.5 ± 29 µM and 217 ± 22.5 µmol min−1 mg−1 protein, respectively.

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Acknowledgements

Bhawna Madan gratefully acknowledges senior research fellowship award from the Council of Scientific and Industrial Research, New Delhi, India. This work was partially supported by a grant from Ministry of Human Resource Development, Government of India to one of the authors (PM).

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  1. Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi, 110016, India

    Bhawna Madan & Prashant Mishra

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  1. Bhawna Madan
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  2. Prashant Mishra
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Correspondence to Prashant Mishra.

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Madan, B., Mishra, P. Co-expression of the lipase and foldase of Pseudomonas aeruginosa to a functional lipase in Escherichia coli . Appl Microbiol Biotechnol 85, 597–604 (2010). https://doi.org/10.1007/s00253-009-2131-4

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  • DOI: https://doi.org/10.1007/s00253-009-2131-4

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