title: Cloning, expression, and characterization of Aureobasidium melanogenum lipase in Pichia pastoris.
abstract: 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.
authors.affiliation: a Program in Biotechnology, Faculty of Science , Chulalongkorn University , Bangkok , Thailand.|b Biofuels by Biocatalysts Research Unit, Faculty of Science, Department of Botany , Chulalongkorn University , Bangkok , Thailand.
authors.forename: Jinaporn
authors.initials: J
authors.lastname: Wongwatanapaiboon
authors.affiliation: c National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA) , Pathum Thani , Thailand.|d Faculty of Science, Center of Excellence for Marine Biotechnology , Chulalongkorn University , Bangkok , Thailand.
authors.forename: Sirawut
authors.initials: S
authors.lastname: Klinbunga
authors.affiliation: e Environmental Research and Management Department , PTT Research and Technology Institute, PTT Public Co. Ltd. , Ayutthaya , Thailand.
authors.forename: Chalermchai
authors.initials: C
authors.lastname: Ruangchainikom
authors.affiliation: e Environmental Research and Management Department , PTT Research and Technology Institute, PTT Public Co. Ltd. , Ayutthaya , Thailand.
authors.forename: Gamgarn
authors.initials: G
authors.lastname: Thummadetsak
authors.affiliation: f Faculty of Engineering, Department of Chemical Engineering , Mahidol University , Nakhon Pathom , Thailand.
authors.forename: Suphang
authors.initials: S
authors.lastname: Chulalaksananukul
authors.affiliation: g Université de Toulouse, INSA, UPS, INP, LISBP , Toulouse , France.|h CNRS, UMR5504 , Toulouse , France.|i INRA, UMR792, Ingénierie des Systèmes Biologiques et des Procédés , Toulouse , France.
authors.forename: Alain
authors.initials: A
authors.lastname: Marty
authors.affiliation: b Biofuels by Biocatalysts Research Unit, Faculty of Science, Department of Botany , Chulalongkorn University , Bangkok , Thailand.|j Faculty of Science, Department of Botany , Chulalongkorn University , Bangkok , Thailand.|k Aquatic Resources Research Institute, Chulalongkorn University , Bangkok , Thailand.
authors.forename: Warawut
authors.initials: W
authors.lastname: Chulalaksananukul
keywords: expression; lipase