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Purification and characterization of a novel organic solvent-tolerant and cold-adapted lipase from Psychrobacter sp. ZY124

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Abstract

By screening 25 different psychrophilic strains isolated from the Arctic habitat, we isolated a strain capable of producing lipase. We identified this strain as Psychrobacter sp. ZY124 based on the amplified 16S rDNA sequence. The lipase, named as Lipase ZC12, produced from the supernatant of Psychrobacter sp. ZY124 cultured at 15 °C was purified to homogeneity by ammonium sulfate precipitation followed by Phenyl Sepharose FF gel hydrophobic chromatography. Based on the obtained amino acid sequence, Lipase ZC12 is classified as a member of the Proteus/psychrophilic subfamily of lipase family I.1; it has a molecular weight of 37.9 kDa. We also determined that the apparent optimum temperature for Lipase ZC12 activity is 40 °C. Lipase ZC12 shows remarkable organic solvent tolerance by remaining more 50% after incubated with 10–90% different organic solvents. In addition, acyl chain esters with C12 or longer were confirmed to be preferable substrates for Lipase ZC12. Lipase ZC12 also shows better stereoselectivity for (R, S)-1-phenylethanol chiral resolution in n-hexane solvent with (S)-1-phenylethanol (eep 92%) and conversion rate (39%) by transesterification reactions. These properties may provide potential applications in biocatalysis and biotransformation in non-aqueous media, such as in detergent, transesterification or esterification and chiral resolution.

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Abbreviations

pNPA:

p-Nitrophenyl acetate

pNPB:

p-Nitrophenyl butyrate

pNPH:

p-Nitrophenyl hexanoate

pNPC:

p-Nitrophenyl caprylate

pNPL:

p-Nitrophenyl laurate

pNPP:

p-Nitrophenyl palmitate

DMSO:

Dimethyl sulfoxide

GC:

Gas chromatography

CD:

Circular dichroism

NCBI:

National Center for Biotechnology Information

ees :

Enantiomeric excess value of the substrate

eep :

Enantiomeric excess value of the product

C :

Conversion ratio

E :

Enantioselectivity

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Acknowledgements

This work was supported by Panjin Campus for Food Science and Technology Research Initiative, Dalian University of Technology.

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Correspondence to Yongming Bao.

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Zhang, Y., Ji, F., Wang, J. et al. Purification and characterization of a novel organic solvent-tolerant and cold-adapted lipase from Psychrobacter sp. ZY124. Extremophiles 22, 287–300 (2018). https://doi.org/10.1007/s00792-018-0997-8

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  • DOI: https://doi.org/10.1007/s00792-018-0997-8

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