title: The thioredoxin reductase-encoding gene ActrxR1 is involved in the cephalosporin C production of Acremonium chrysogenum in methionine-supplemented medium.
abstract: The thioredoxin system including thioredoxin and thioredoxin reductase (TrxR) is used for oxidative stress defenses in fungi. Based on the genomic sequence, a thioredoxin reductase-encoding gene (ActrxR1) was isolated from Acremonium chrysogenum CGMCC3.3795. Like other TrxRs, AcTrxR1 contains FAD binding domain, Redox domain, and NADPH binding domain. Disruption of ActrxR1 in A. chrysogenum led to the formation of smaller colonies and hyphal swelling in Tryptic soy agar (TSA). In chemically defined medium, the spore germination of ActrxR1 disruption mutant was strongly inhibited, which was recovered by the addition of DL-methionine. The disruption mutant grew slowly on TSA compared with the wild-type strain, but it did not show to be more sensitive to exogenous hydrogen peroxide or menadione. In defined medium of fermentation supplemented with DL-methionine, the ActrxR1 disruption mutant grew normally, and its cephalosporin C production increased by about onefold compared with the wild type (73 μg/ml for wild-type strain and 136 μg/ml for the mutant at 5 days of fermentation). Real-time polymerase chain reaction (RT-PCR) showed that the transcriptional levels of pcbC, cefEF, and cefG were obviously enhanced in the ActrxR1 mutant at the early stage of fermentation. These results indicate that ActrxR1 is required for the normal growth of A. chrysogenum and related with cephalosporin C production in methionine-supplemented medium.
authors.affiliation: Institute of Modern Biopharmaceuticals, School of Pharmaceutical Sciences, Southwest University, Beibei, Chongqing 400716, China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Liang-Kun
authors.initials: LK
authors.lastname: Long
authors.forename: Yang
authors.initials: Y
authors.lastname: An
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Xinxin
authors.initials: X
authors.lastname: Xu
authors.forename: Chang-Hua
authors.initials: CH
authors.lastname: Hu
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
mesh_terms: D000164:Acremonium / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D001665:Binding Sites; D002511:Cephalosporins / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D025301:Hyphae / Q000254:growth & development; D008715:Methionine / Q000378:metabolism*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013172:Spores, Fungal / Q000254:growth & development; D013880:Thioredoxin-Disulfide Reductase / Q000378:metabolism*
chemical_list: D002511:Cephalosporins; D003470:Culture Media; D004271:DNA, Fungal; C025163:cephalosporin C; D008715:Methionine; D013880:Thioredoxin-Disulfide Reductase