RRC ID 54502
Author Polburee P, Ohashi T, Tsai YY, Sumyai T, Lertwattanasakul N, Limtong S, Fujiyama K.
Title Molecular cloning and overexpression of DGA1, an acyl-CoA-dependent diacylglycerol acyltransferase, in the oleaginous yeast Rhodosporidiobolus fluvialis DMKU-RK253.
Journal Microbiology (Reading)
Abstract Triacylglycerol (TAG) is a major component of lipid storage in yeast. The acyl CoA: diacylgycerol acyltransferase (DGAT) that catalyzes the final and rate-limiting step in the production of TAG is rather interesting. Consequently, cloning and analysis of the gene-encoding TAG synthase, diacylglycerol acyltransferase gene (DGA1), of the oleaginous yeast Rhodosporidiobolus fluvialis DMKU-RK253 were undertaken. Analysis of the deduced amino acid sequence of DGA1 from R. fluvialis DMKU-RK253 (RfDGA1) showed similarity with the acyl-CoA:diacylglycerol acyltransferase 2 (DGAT2) from other organisms. The cDNA of RfDGA1 was cloned into the yeast expression vector pYES2 and heterologously overexpressed in Saccharomyces cerevisiae. One of the transformants showed a 1.6-fold increase in lipid content compared with the wild-type strain harbouring the pYES2 empty vector. Furthermore, DGA1 overexpression in R. fluvialis DMKU-RK253 resulted in a 2.5-fold increase in lipid content when compared with the wild-type strain, and no significant differences in fatty acid composition were observed between RfDGA1-overexpressed and wild-type strains. Taken together, our results supported our hypothesis that the RfDGA1 is a genetic factor that can be used for the development of a strain with improved lipid accumulation capabilities.
Volume 164(1)
Pages 1-10
Published 2018-1
DOI 10.1099/mic.0.000584
PMID 29182511
MeSH Amino Acid Sequence Basidiomycota / classification Basidiomycota / enzymology* Basidiomycota / genetics Basidiomycota / metabolism Biomass Cloning, Molecular Diacylglycerol O-Acyltransferase / genetics* Diacylglycerol O-Acyltransferase / metabolism Fatty Acids / analysis Fungal Proteins / genetics* Fungal Proteins / metabolism Gene Expression* Glycerol / metabolism Lipid Metabolism / genetics* Metabolic Engineering Phylogeny Saccharomyces cerevisiae / enzymology Saccharomyces cerevisiae / genetics Sequence Homology, Amino Acid Triglycerides / biosynthesis
IF 1.922
Times Cited 3
Resource
Yeast BY23849