Open AccessCloning and functional identification of C‐4 methyl sterol oxidase genes from the penicillin‐producing fungus Penicillium chrysogenum
Author(s) -
Wang FuQiang,
Zhao Ying,
Dai Meng,
Liu Jing,
Zheng GuiZhen,
Ren ZhiHong,
He JianGong
Publication year - 2008
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2008.01294.x
Subject(s) - penicillium chrysogenum , biology , ergosterol , sterol , complementation , open reading frame , gene , biochemistry , saccharomyces cerevisiae , penicillin , microbiology and biotechnology , peptide sequence , phenotype , antibiotics , cholesterol
Two C‐4 methyl sterol oxidase genes ( Pcerg25A and Pcerg25B ) that are involved in ergosterol biosynthesis have been cloned from the penicillin‐producing fungus Penicillium chrysogenum . cDNAs of both Pcerg25 A and Pcerg25B have an ORF 885 bp in length, encoding a peptide of 295 residues. The deduced amino acid sequences of PcErg25A and PcErg25B show 86% identity, and have high identities to the characterized C‐4 methyl sterol oxidases from Candida albicans and Saccharomyces cerevisiae . The function of Pcerg25A and Pcerg25B was identified by complementation of a yeast erg25 ‐deficient strain. Pcerg25A is located in the DNA region containing the penicillin gene cluster, and thus its copy number is dependent on the patterns of the cluster region. Up to eight copies of Pcerg25A were found in the high‐productivity strain NCPC 10086. By contrast, Pcerg25B was present in just a single copy in all tested P. chrysogenum genomes. Differences in the transcript level of either Pcerg25A or Pcerg25B were observed in different P. chrysogenum strains by real‐time quantitative reverse transcriptase PCR analysis.