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Molecular characterization and expression analysis of GlHMGS, a gene encoding hydroxymethylglutaryl-CoA synthase from Ganoderma lucidum (Ling-zhi) in ganoderic acid biosynthesis pathway

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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.

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Abbreviations

HMGS:

Hydroxymethylglutaryl-CoA synthase

GA:

Ganoderic acid

ORF:

Open reading frame

SA:

Salicylic acid

ABA:

Abscisic acid

MeJA:

Methyl jasmonate

PDA:

Potato dextrose agar

PDB:

Potato dextrose broth

SEFA PCR:

Self-Formed Adaptor PCR

ATMT:

Agrobacterium tumefaciens-mediated transformation

GPD:

Glyceraldehyde-3-phosphate dehydrogenase

HPH:

Hygromycin phosphotransferase

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Acknowledgments

This work was supported financially by the National Natural Science Foundation of China (project No. 30970042 and No. J1210056), the Fundamental Research Funds for the Central Universities (project No. KYZ201121), the Chinese National Key Technology R&D Program (Project No. 2012BAD36B05), the Science & Technology Pillar Program of Jiangsu Province, China (Project No. BE2011312), and the Program for scientific innovation research of college graduate in Jiangsu Province (project No. CXZZ11_0648 and No. CXZZ12_0265).

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Authors and Affiliations

  1. 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

    Ang Ren, Xiang Ouyang, Liang Shi, Ai-Liang Jiang, Da-Shuai Mu, Meng-Jiao Li, Qin Han & Ming-Wen Zhao

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  1. Ang Ren
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  2. Xiang Ouyang
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  3. Liang Shi
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Correspondence to Ming-Wen Zhao.

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Ren, A., Ouyang, X., Shi, L. et al. Molecular characterization and expression analysis of GlHMGS, a gene encoding hydroxymethylglutaryl-CoA synthase from Ganoderma lucidum (Ling-zhi) in ganoderic acid biosynthesis pathway. World J Microbiol Biotechnol 29, 523–531 (2013). https://doi.org/10.1007/s11274-012-1206-z

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  • DOI: https://doi.org/10.1007/s11274-012-1206-z

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