title: Identification and characterization of a type-2 diacylglycerol acyltransferase (DGAT2) from Rhodosporidium diobovatum.
abstract: Triacylglycerols (TAGs), synthesized in the microsomal membranes of eukaryotes, serve as a primary storage form of carbon and energy in microorganisms. For this reason, TAGs produced by organisms have great potential to become biofuels and facilitate researchers to look for alternative renewable sources of energy. The present study describes the identification and functional characterization of a type-2 diacylglycerol acyltransferase from Rhodosporidium diobovatum, designated as RdDGAT, which catalyzed the final step of TAG synthesis. A full-length cDNA clone for RdDGAT was obtained, and its biological activity was proven by being expressed in a Saccharomyces cerevisiae quadruple mutant that was defective in TAG synthesis. Enzymatic assays were performed and finally the existence of TAGs in the transformed Saccharomyces cerevisiae quadruple mutant was determined using the method of thin-layer chromatography. Substrate preference experiments revealed that RdDGAT preferred unsaturated fatty acids over saturated ones. Through further analysis, we assume that the evolution and expression characteristics of the RdDGAT gene perhaps is the result of adaption to its oligotrophic and cold living environment.
authors.affiliation: The Key Lab of Microbial Diversity Research and Application of Hebei Province; Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education; College of Life Sciences, Hebei University, Baoding, 071002, China, chenzhihuanasdfg@163.com.
authors.forename: Zhihuan
authors.initials: Z
authors.lastname: Chen
authors.forename: Pengyan
authors.initials: P
authors.lastname: Liu
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Liu
authors.forename: Hui
authors.initials: H
authors.lastname: Tang
authors.forename: Yunping
authors.initials: Y
authors.lastname: Chen
authors.forename: Liping
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D016000:Cluster Analysis; D018076:DNA, Complementary; D051048:Diacylglycerol O-Acyltransferase / Q000235:genetics* / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D010802:Phylogeny; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D005227:Fatty Acids; D051048:Diacylglycerol O-Acyltransferase