Publication: Cloning, expression, and characterization of Aureobasidium melanogenum lipase in Pichia pastoris
Issued Date
2016-01-01
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ISSN
13476947
09168451
09168451
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2-s2.0-84992723936
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Mahidol University
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SCOPUS
Bibliographic Citation
Bioscience, Biotechnology and Biochemistry. Vol.80, No.11 (2016), 2231-2240
Suggested Citation
Jinaporn Wongwatanapaiboon, Sirawut Klinbunga, Chalermchai Ruangchainikom, Gamgarn Thummadetsak, Suphang Chulalaksananukul, Alain Marty, Warawut Chulalaksananukul Cloning, expression, and characterization of Aureobasidium melanogenum lipase in Pichia pastoris. Bioscience, Biotechnology and Biochemistry. Vol.80, No.11 (2016), 2231-2240. doi:10.1080/09168451.2016.1206809 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/43290
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Title
Cloning, expression, and characterization of Aureobasidium melanogenum lipase in Pichia pastoris
Abstract
© 2016 Japan Society for Bioscience, Biotechnology, and Agrochemistry. cDNA of Aureobasidium melanogenum lipase comprises 1254 bp encoding 417 amino acids, whereas genomic DNA of lipase comprises 1311 bp with one intron (57 bp). The lipase gene contains a putative signal peptide encoding 26 amino acids. The A. melanogenum lipase gene was successfully expressed in Pichia pastoris. Recombinant lipase in an inducible expression system showed the highest lipase activity of 3.8 U/mL after six days of 2% v/v methanol induction. The molecular mass of purified recombinant lipase was estimated as 39 kDa using SDS-PAGE. Optimal lipase activity was observed at 35-37 °C and pH 7.0 using p-nitrophenyl laurate as the substrate. Lipase activity was enhanced by Mg2+, Mn2+, Li+, Ca2+, Ni2+, CHAPS, DTT, and EDTA and inhibited by Hg2+, Ag+, SDS, Tween 20, and Triton X-100. The addition of 10% v/v acetone, DMSO, p-xylene, and octanol increased lipase activity, whereas that of propanol and butanol strongly inhibited it.