Cloning and Molecular Analysis of Extracellular Lipase Gene From Native and Mutant Yarrowia lipolytica Strains

Abstract

Background: The lipase enzyme of the Yarrowia lipolytica yeast is an important enzyme in pharmaceutical, food and environmental biotechnology. Objectives: The aims of the study were cloning and molecular analysis of extracellular lipase genes from DSM3286 native and U6 mutant strains of Y. lipolytica in Escherichia coli. Materials and Methods: LIP2 gene of Y. lipolytica DSM3286 and its mutant Y. lipolytica U6 were isolated and cloned in E. coli using the appropriate vector. The native and mutant extracellular lipase ORFs were sequenced for further analyses and finding the differences in the nucleotide sequences. Results: Two nucleotide substitutions were found in the extracellular lipase LIP2 ORFs of Y. lipolytica U6 mutant strain compared to the wild-type strain. In this study, the LIP2 ORF region genetic changes were found in the industrial and high-level extracellular lipase producer Y. lipolytica mutant strain. Conclusions: Results of this study could be used to find suitable targets for inverse metabolic engineering and site-directed mutagenesis to achieve stable and high-level extracellular lipase production in Y. lipolytica and other yeasts.