Knockout of the SREBP system increases production of the polyketide FR901512 in filamentous fungal sp. No. 14919 and lovastatin in Aspergillus terreus ATCC20542

Hiroya Itoh, Ai Miura, Makoto Matsui, Takayuki Arazoe, Keiji Nishida, Toshitaka Kumagai, Masanori Arita, Koichi Tamano, Masayuki Machida, Takashi Shibata

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

In the production of useful microbial secondary metabolites, the breeding of strains is generally performed by random mutagenesis. However, because random mutagenesis introduces many mutations into genomic DNA, the causative mutations leading to increased productivity are mostly unknown. Therefore, although gene targeting is more efficient for breeding than random mutagenesis, it is difficult to apply. In this study, a wild-type strain and randomly mutagenized strains of fungal sp. No. 14919, a filamentous fungus producing the HMG-CoA reductase inhibitor polyketide FR901512, were subjected to point mutation analysis based on whole genome sequencing. Among the mutated genes found, mutation of the sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP) had a positive effect on increasing FR901512 productivity. By complementing the SCAP gene in the SCAP-mutated strain, productivity was decreased to the level of the SCAP-intact strain. Conversely, when either the SCAP or SREBP gene was deleted, the productivity was significantly increased. By genomic transcriptional analysis, the expression levels of three enzymes in the ergosterol biosynthesis pathway were shown to be decreased by SCAP mutation. These findings led to the hypothesis that raw materials of polyketides, such as acetyl-CoA and malonyl-CoA, became more available for FR901512 biosynthesis due to depression in sterol biosynthesis caused by knockout of the SREBP system. This mechanism was confirmed in Aspergillus terreus producing the polyketide lovastatin, which is structurally similar to FR901512. Thus, knockout of the SREBP system should be considered significant for increasing the productivities of polyketides, such as HMG-CoA reductase inhibitors, by filamentous fungi.

Original languageEnglish
Pages (from-to)1393-1405
Number of pages13
JournalApplied Microbiology and Biotechnology
Volume102
Issue number3
DOIs
Publication statusPublished - 1 Feb 2018

Fingerprint

Sterol Regulatory Element Binding Proteins
Polyketides
Lovastatin
Aspergillus
Mutagenesis
Hydroxymethylglutaryl-CoA Reductase Inhibitors
Mutation
Breeding
Fungi
Malonyl Coenzyme A
Genes
Ergosterol
Acetyl Coenzyme A
Gene Targeting
Sterols
Point Mutation
SREBP cleavage-activating protein
FR901512
Genome
DNA

Keywords

  • FR091512
  • Fungal sp. no. 14919
  • HMG-CoA reductase inhibitor
  • Polyketide synthase
  • SREBP
  • Secondary metabolism

Cite this

Itoh, Hiroya ; Miura, Ai ; Matsui, Makoto ; Arazoe, Takayuki ; Nishida, Keiji ; Kumagai, Toshitaka ; Arita, Masanori ; Tamano, Koichi ; Machida, Masayuki ; Shibata, Takashi. / Knockout of the SREBP system increases production of the polyketide FR901512 in filamentous fungal sp. No. 14919 and lovastatin in Aspergillus terreus ATCC20542. In: Applied Microbiology and Biotechnology. 2018 ; Vol. 102, No. 3. pp. 1393-1405.
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Knockout of the SREBP system increases production of the polyketide FR901512 in filamentous fungal sp. No. 14919 and lovastatin in Aspergillus terreus ATCC20542. / Itoh, Hiroya; Miura, Ai; Matsui, Makoto; Arazoe, Takayuki; Nishida, Keiji; Kumagai, Toshitaka; Arita, Masanori; Tamano, Koichi; Machida, Masayuki; Shibata, Takashi.

In: Applied Microbiology and Biotechnology, Vol. 102, No. 3, 01.02.2018, p. 1393-1405.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Knockout of the SREBP system increases production of the polyketide FR901512 in filamentous fungal sp. No. 14919 and lovastatin in Aspergillus terreus ATCC20542

AU - Itoh, Hiroya

AU - Miura, Ai

AU - Matsui, Makoto

AU - Arazoe, Takayuki

AU - Nishida, Keiji

AU - Kumagai, Toshitaka

AU - Arita, Masanori

AU - Tamano, Koichi

AU - Machida, Masayuki

AU - Shibata, Takashi

PY - 2018/2/1

Y1 - 2018/2/1

N2 - In the production of useful microbial secondary metabolites, the breeding of strains is generally performed by random mutagenesis. However, because random mutagenesis introduces many mutations into genomic DNA, the causative mutations leading to increased productivity are mostly unknown. Therefore, although gene targeting is more efficient for breeding than random mutagenesis, it is difficult to apply. In this study, a wild-type strain and randomly mutagenized strains of fungal sp. No. 14919, a filamentous fungus producing the HMG-CoA reductase inhibitor polyketide FR901512, were subjected to point mutation analysis based on whole genome sequencing. Among the mutated genes found, mutation of the sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP) had a positive effect on increasing FR901512 productivity. By complementing the SCAP gene in the SCAP-mutated strain, productivity was decreased to the level of the SCAP-intact strain. Conversely, when either the SCAP or SREBP gene was deleted, the productivity was significantly increased. By genomic transcriptional analysis, the expression levels of three enzymes in the ergosterol biosynthesis pathway were shown to be decreased by SCAP mutation. These findings led to the hypothesis that raw materials of polyketides, such as acetyl-CoA and malonyl-CoA, became more available for FR901512 biosynthesis due to depression in sterol biosynthesis caused by knockout of the SREBP system. This mechanism was confirmed in Aspergillus terreus producing the polyketide lovastatin, which is structurally similar to FR901512. Thus, knockout of the SREBP system should be considered significant for increasing the productivities of polyketides, such as HMG-CoA reductase inhibitors, by filamentous fungi.

AB - In the production of useful microbial secondary metabolites, the breeding of strains is generally performed by random mutagenesis. However, because random mutagenesis introduces many mutations into genomic DNA, the causative mutations leading to increased productivity are mostly unknown. Therefore, although gene targeting is more efficient for breeding than random mutagenesis, it is difficult to apply. In this study, a wild-type strain and randomly mutagenized strains of fungal sp. No. 14919, a filamentous fungus producing the HMG-CoA reductase inhibitor polyketide FR901512, were subjected to point mutation analysis based on whole genome sequencing. Among the mutated genes found, mutation of the sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP) had a positive effect on increasing FR901512 productivity. By complementing the SCAP gene in the SCAP-mutated strain, productivity was decreased to the level of the SCAP-intact strain. Conversely, when either the SCAP or SREBP gene was deleted, the productivity was significantly increased. By genomic transcriptional analysis, the expression levels of three enzymes in the ergosterol biosynthesis pathway were shown to be decreased by SCAP mutation. These findings led to the hypothesis that raw materials of polyketides, such as acetyl-CoA and malonyl-CoA, became more available for FR901512 biosynthesis due to depression in sterol biosynthesis caused by knockout of the SREBP system. This mechanism was confirmed in Aspergillus terreus producing the polyketide lovastatin, which is structurally similar to FR901512. Thus, knockout of the SREBP system should be considered significant for increasing the productivities of polyketides, such as HMG-CoA reductase inhibitors, by filamentous fungi.

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