title: Overexpression of the homologous lanosterol synthase gene in ganoderic acid biosynthesis in Ganoderma lingzhi.
abstract: Ganoderic acids (GAs) in Ganoderma lingzhi exhibit anticancer and antimetastatic activities. GA yields can be potentially improved by manipulating G. lingzhi through genetic engineering. In this study, a putative lanosterol synthase (LS) gene was cloned and overexpressed in G. lingzhi. Results showed that its overexpression (OE) increased the ganoderic acid (GA) content and the accumulation of lanosterol and ergosterol in a submerged G. lingzhi culture. The maximum contents of GA-O, GA-Mk, GA-T, GA-S, GA-Mf, and GA-Me in transgenic strains were 46.6 ± 4.8, 24.3 ± 3.5, 69.8 ± 8.2, 28.9 ± 1.4, 15.4 ± 1.2, and 26.7 ± 3.1 μg/100 mg dry weight, respectively, these values being 6.1-, 2.2-, 3.2-, 4.8-, 2.0-, and 1.9-times higher than those in wild-type strains. In addition, accumulated amounts of lanosterol and ergosterol in transgenic strains were 2.3 and 1.4-fold higher than those in the control strains, respectively. The transcription level of LS was also increased by more than five times in the presence of the G. lingzhi glyceraldehyde-3-phosphate dehydrogenase gene promoter, whereas transcription levels of 3-hydroxy-3-methylglutaryl coenzyme A enzyme and squalene synthase did not change significantly in transgenic strains. This study demonstrated that OE of the homologous LS gene can enhance lanosterol accumulation. A large precursor supply promotes GA biosynthesis.
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: De-Huai
authors.initials: DH
authors.lastname: Zhang
authors.affiliation: Faculty of Science, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Na
authors.initials: N
authors.lastname: Li
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Xuya
authors.initials: X
authors.lastname: Yu
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhao
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China. Electronic address: xjuwei@163.com.
authors.forename: Jun-Wei
authors.initials: JW
authors.lastname: Xu
mesh_terms: D004875:Ergosterol / Q000032:analysis; D032902:Ganoderma / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000378:metabolism; D019751:Intramolecular Transferases / Q000235:genetics / Q000378:metabolism*; D007810:Lanosterol / Q000032:analysis; D030821:Plants, Genetically Modified; D020940:Reishi / Q000737:chemistry; D014315:Triterpenes / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
chemical_list: D014315:Triterpenes; C556862:ganoderic acid; C572162:ganoderic acid Mf; D007810:Lanosterol; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D019751:Intramolecular Transferases; C021924:lanosterol synthase; D004875:Ergosterol
keywords: Fermentation; Ganoderma lucidum; Genetic engineering; Medicinal mushroom