Further enhancement of FR901469 productivity by co-overexpression of cpcA, a cross-pathway control gene, and frbF in fungal sp. No. 11243.
FR901469 is a secondary metabolite with antifungal activity, produced by fungal sp. No. 11243. In our previous study, we constructed the frbF overexpression mutant (TFH2-2) from the wild-type strain. FR901469 productivity of TFH2-2 was 3.4 times higher than that of the wild-type strain. To further enhance FR901469 productivity in TFH2-2, we attempted to find genes from the genome that limited the productivity as bottlenecks in this study. Based on both correlation analysis of gene expression level against FR901469 productivity and genome annotation information, the cross-pathway control gene A (cpcA) was most predicted as the bottleneck. The cpcA and frbF co-overexpression mutant named TFCH3 was then constructed from TFH2-2. As a result, FR901469 productivity of TFCH3 was enhanced at 1.8 times higher than that of TFH2-2. Transcriptome analysis revealed that many genes involved in amino acid biosynthesis and encoding tRNA ligases were significantly upregulated in TFCH3, which implied increase of amino acids as the substrates of FR901469 would be a reason of further productivity enhancement.
Biotechnology Labs, Astellas Pharma Inc., 5-2-3 Tokodai, Tsukuba-shi, Ibaraki 300-2698, Japan; Technology Research Association of Highly Efficient Gene Design (TRAHED), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan. Electronic address: makoto.matsui@astellas.com.
Makoto
M
Matsui
Biotechnology Labs, Astellas Pharma Inc., 5-2-3 Tokodai, Tsukuba-shi, Ibaraki 300-2698, Japan; Technology Research Association of Highly Efficient Gene Design (TRAHED), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan.
Tatsuya
T
Yokoyama
Technology Research Association of Highly Efficient Gene Design (TRAHED), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan.
Kaoru
K
Nemoto
Fermlab, Inc., 4-3-1-913 Shirakawa, Koto-ku, Tokyo 135-0021, Japan.
Toshitaka
T
Kumagai
Technology Research Association of Highly Efficient Gene Design (TRAHED), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan.
Koichi
K
Tamano
Technology Research Association of Highly Efficient Gene Design (TRAHED), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan; Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan.
Masayuki
M
Machida
Biotechnology Labs, Astellas Pharma Inc., 5-2-3 Tokodai, Tsukuba-shi, Ibaraki 300-2698, Japan; Technology Research Association of Highly Efficient Gene Design (TRAHED), 17-2-1 Higashi-Nijo, Tsukisamu, Toyohira-ku, Sapporo, Hokkaido 062-8517, Japan.
Takashi
T
Shibata
D000935:Antifungal Agents / Q000378:metabolism*; D047630:Depsipeptides / Q000096:biosynthesis*; D005658:Fungi / Q000235:genetics*; D015870:Gene Expression; D005800:Genes, Fungal / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D009154:Mutation; D055666:Lipopeptides
D000935:Antifungal Agents; D047630:Depsipeptides; C417112:FR 901469; D055666:Lipopeptides
FR901469; Fungal sp. No. 11243; Secondary metabolite; Transcriptome; cpcA