High-level production and characterisation of the recombinant thermostable lipase of Geobacillus thermoleovorans in Pichia methanolica

Abstract

The thermostable lipase-encoding gene from Geobacillus thermoleovorans has been cloned and expressed in the Pichia methanolica under the control of the alcohol oxidase (AUG1) promoter which was followed by the Saccharomyces cerevisiae α-factor signal peptide. The α-factor signal peptide efficiently directed the secretion of lipase from the recombinant yeast cell. Recombinant lipase was produced in a biological active form by P. methanolica. In shaken flask cultivations the expression levels (90 U/ml) were 30-fold higher than in the wild-type strain. The recombinant pure lipase exhibited optimal activity at 60 °C and pH 8.0.