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Cloning, expression, and characterization of Aureobasidium melanogenum lipase in Pichia pastoris.

Authors :
Wongwatanapaiboon, Jinaporn
Klinbunga, Sirawut
Ruangchainikom, Chalermchai
Thummadetsak, Gamgarn
Chulalaksananukul, Suphang
Marty, Alain
Chulalaksananukul, Warawut
Source :
Bioscience, Biotechnology & Biochemistry. Nov2016, Vol. 80 Issue 11, p2231-2240. 10p.
Publication Year :
2016

Abstract

cDNA ofAureobasidium melanogenumlipase 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. TheA. melanogenumlipase gene was successfully expressed inPichia 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 usingp-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. Lipase activity ofAureobasidium melanogenumlipase expressed inPichia pastorisusing inducible and constitutive expression system. [ABSTRACT FROM PUBLISHER]

Details

Language :
English
ISSN :
09168451
Volume :
80
Issue :
11
Database :
Academic Search Index
Journal :
Bioscience, Biotechnology & Biochemistry
Publication Type :
Academic Journal
Accession number :
118355605
Full Text :
https://doi.org/10.1080/09168451.2016.1206809