Improvement in the Organic Solvent Stability of <i>Pseudomonas</i> Lipase by Random Mutation<sup>a</sup>

“…For example, a lipase variant from Pseudomonas sp. KWI-56, which was found to be 40% more stable in 80% (v/v) DMSO than wild type, had only a single surface residue mutation (V304A; Nakano et al 1998). Similarly, a G157R mutation in Pseudomonas aeruginosa LST-03 lipase introduced additional bonds that promoted salt bridge and H-bond formation.…”

Section: Enhanced Stabilising Interactionsmentioning

confidence: 99%

Solvent stable microbial lipases: current understanding and biotechnological applications

Priyanka¹,

Tan²,

Kinsella³

et al. 2018

Biotechnol Lett

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Objective: This review examines on our current understanding of microbial lipase solvent tolerance, with a specific focus on the molecular strategies employed to improve lipase stability in a non-aqueous environment. Results: It provides an overview of known solvent tolerant lipases and of approaches to improving solvent stability such as; enhancing stabilising interactions, modification of residue flexibility and surface charge alteration. It shows that judicious selection of lipase source supplemented by appropriate enzyme stabilisation, can lead to a wide application spectrum for lipases. Conclusion: Organic solvent stable lipases are, and will continue to be, versatile and adaptable biocatalytic workhorses commonly employed for industrial applications in the food, pharmaceutical and green manufacturing industries.

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Enhancement of the organic solvent‐stability of the LST‐03 lipase by directed evolution

Kawata

Ogino

2009

Biotechnology Progress

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LST-03 lipase from an organic solvent-tolerant Pseudomonas aeruginosa LST-03 has high stability and activity in the presence of various organic solvents. In this research, enhancement of organic solvent-stability of LST-03 lipase was attempted by directed evolution. The structural gene of the LST-03 lipase was amplified by the error prone-PCR method. Organic solvent-stability of the mutated lipases was assayed by formation of a clear zone of agar which contained dimethyl sulfoxide (DMSO) and tri-n-butyrin and which overlaid a plate medium. And the organic solvent-stability was also confirmed by measuring the half-life of activity in the presence of DMSO. Four mutated enzymes were selected on the basis of their high organic solvent-stability in the presence of DMSO. The organic solvent-stabilities of mutated LST-03 lipase in the presence of various organic solvents were measured and their mutated amino acid residues were identified. The half-lives of the LST-03-R65 lipase in the presence of cyclohexane and n-decane were about 9 to 11-fold longer than those of the wild-type lipase, respectively. Some substituted amino acid residues of mutated LST-03 lipases have been located at the surface of the enzyme molecules, while some other amino acid residues have been changed from neutral to basic residues.

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Improvement in the Organic Solvent Stability of Pseudomonas Lipase by Random Mutation^a

Abstract: NAKANO et al.: P. LIPASE 433 FIGURE 3. Three-dimensional structure of the P. sp. KWI-56 lipase, based on homology modeling. The side chains of amino acid residues at mutated positions are shown. The catalytic triad is represented as a CPK model.

Cited by 5 publications

References 5 publications

An organic solvent and thermally stable lipase from Burkholderia ambifaria YCJ01: Purification, characteristics and application for chiral resolution of mandelic acid

An organic solvent and thermally stable lipase from Burkholderia ambifaria YCJ01: Purification, characteristics and application for chiral resolution of mandelic acid

Solvent stable microbial lipases: current understanding and biotechnological applications

Enhancement of the organic solvent‐stability of the LST‐03 lipase by directed evolution

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Improvement in the Organic Solvent Stability of Pseudomonas Lipase by Random Mutationa

Abstract: NAKANO et al.: P. LIPASE 433 FIGURE 3. Three-dimensional structure of the P. sp. KWI-56 lipase, based on homology modeling. The side chains of amino acid residues at mutated positions are shown. The catalytic triad is represented as a CPK model.

Cited by 5 publications

References 5 publications

An organic solvent and thermally stable lipase from Burkholderia ambifaria YCJ01: Purification, characteristics and application for chiral resolution of mandelic acid

An organic solvent and thermally stable lipase from Burkholderia ambifaria YCJ01: Purification, characteristics and application for chiral resolution of mandelic acid

Solvent stable microbial lipases: current understanding and biotechnological applications

Enhancement of the organic solvent‐stability of the LST‐03 lipase by directed evolution

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Improvement in the Organic Solvent Stability of Pseudomonas Lipase by Random Mutation^a