title: Molecular characterization and expression analysis of GlHMGS, a gene encoding hydroxymethylglutaryl-CoA synthase from Ganoderma lucidum (Ling-zhi) in ganoderic acid biosynthesis pathway.
abstract: A hydroxymethylglutaryl-CoA synthase gene, designated as GlHMGS (GenBank accession No. JN391469) involved in ganoderic acid (GA) biosynthesis pathway was cloned from Ganoderma lucidum. The full-length cDNA of GlHMGS (GenBank accession No. JN391468) was found to contain an open reading frame of 1,413 bp encoding a polypeptide of 471 amino acid residues. The deduced amino acid sequence of GlHMGS shared high homology with other known hydroxymethylglutaryl-CoA synthase (HMGS) enzymes. In addition, functional complementation of GlHMGS in a mutant yeast strain YSC1021 lacking HMGS activity demonstrated that the cloned cDNA encodes a functional HMGS. A 1,561 bp promoter sequence was isolated and its putative regulatory elements and potential specific transcription factor binding sites were analyzed. GlHMGS expression profile analysis revealed that salicylic acid, abscisic acid and methyl jasmonate up-regulated GlHMGS transcript levels over the control. Further expression analysis revealed that the developmental stage and carbon source had significant effects on GlHMGS transcript levels. GlHMGS expression peaked on day 16 before decreasing with prolonged culture time. The highest mRNA level was observed when the carbon source was maltose. Overexpression of GlHMGS enhanced GA content in G. lucidum. This study provides useful information for further studying this gene and on its function in the ganoderic acid biosynthetic pathway in G. lucidum.
authors.affiliation: Microbiology Department, Key Laboratory for Microbiological Engineering of the Agricultural Environment, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Weigang No. 1, Nanjing 210095, Jiangsu, People's Republic of China.
authors.forename: Ang
authors.initials: A
authors.lastname: Ren
authors.forename: Xiang
authors.initials: X
authors.lastname: Ouyang
authors.forename: Liang
authors.initials: L
authors.lastname: Shi
authors.forename: Ai-Liang
authors.initials: AL
authors.lastname: Jiang
authors.forename: Da-Shuai
authors.initials: DS
authors.lastname: Mu
authors.forename: Meng-Jiao
authors.initials: MJ
authors.lastname: Li
authors.forename: Qin
authors.initials: Q
authors.lastname: Han
authors.forename: Ming-Wen
authors.initials: MW
authors.lastname: Zhao
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D053898:Biosynthetic Pathways; D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004365:Drugs, Chinese Herbal; D015966:Gene Expression Regulation, Fungal; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D020940:Reishi / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014315:Triterpenes / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D018076:DNA, Complementary; D004365:Drugs, Chinese Herbal; D014315:Triterpenes; C556862:ganoderic acid; D006904:Hydroxymethylglutaryl-CoA Synthase