Poly-γ-glutamic acid production of Bacillus subtilis (natto) in the absence of DegQ: A gain-of-function mutation in yabJ gene

Le Thi Thu Hong, Tsuyoshi Hachiya, Sumitaka Hase, Yuh Shiwa, Hirofumi Yoshikawa, Yasubumi Sakakibara, Sy Le Thanh Nguyen, Keitarou Kimura

Research output: Contribution to journalArticle

Abstract

Poly-γ-glutamic acid (γPGA) production by Bacillus subtilis is regulated by the quorum sensing system where DegQ transmits the cell density signal to a DNA-binding protein DegU. A mutation suppressing the γPGA-negative phenotype of degQ gene knock-out mutant (ΔdegQ) was identified through whole genome sequencing. The mutation conferred an amino acid substitution of Ser103 to phenylalanine (S103F) in yabJ that belongs to the highly conserved YjgF/YER057c/UK114 family. Genetic experiments including LacZ-fusion assay of γPGA synthetic operon confirmed that the suppressor mutation (yabJS103F) was responsible for the recovery of γPGA production. The yabJ itself was not essential for the γPGA production and the mutant allele enabled γPGA production of the ΔdegQ strain even in the presence of wild type yabJ. Thus, yabJS103F was a dominant positive allele. degU-lacZ fusion gene was hyper-expressed in cells carrying the yabJS103F, but disruption of yabJ did not affect the transcription level of the degU-lacZ. These observations suggested that YabJ acquired a function to stimulate expression of degU by the S103F mutation which is involved in the regulation of γPGA synthesis.

Original languageEnglish
JournalJournal of Bioscience and Bioengineering
DOIs
Publication statusPublished - 2019 Jan 1

Fingerprint

Bacilli
Bacillus subtilis
Glutamic Acid
Genes
Mutation
Acids
Fusion reactions
Alleles
Genetic Suppression
Quorum Sensing
Gene Knockout Techniques
Lac Operon
DNA-Binding Proteins
Amino Acid Substitution
Transcription
Operon
Phenylalanine
Amino acids
Assays
DNA

Keywords

  • Bacillus subtilis
  • DegQ
  • Gain-of-function mutation
  • Poly-γ-glutamic acid
  • YabJ
  • YjgF/YER057c/UK114

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Poly-γ-glutamic acid production of Bacillus subtilis (natto) in the absence of DegQ : A gain-of-function mutation in yabJ gene. / Hong, Le Thi Thu; Hachiya, Tsuyoshi; Hase, Sumitaka; Shiwa, Yuh; Yoshikawa, Hirofumi; Sakakibara, Yasubumi; Nguyen, Sy Le Thanh; Kimura, Keitarou.

In: Journal of Bioscience and Bioengineering, 01.01.2019.

Research output: Contribution to journalArticle

Hong, Le Thi Thu ; Hachiya, Tsuyoshi ; Hase, Sumitaka ; Shiwa, Yuh ; Yoshikawa, Hirofumi ; Sakakibara, Yasubumi ; Nguyen, Sy Le Thanh ; Kimura, Keitarou. / Poly-γ-glutamic acid production of Bacillus subtilis (natto) in the absence of DegQ : A gain-of-function mutation in yabJ gene. In: Journal of Bioscience and Bioengineering. 2019.
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abstract = "Poly-γ-glutamic acid (γPGA) production by Bacillus subtilis is regulated by the quorum sensing system where DegQ transmits the cell density signal to a DNA-binding protein DegU. A mutation suppressing the γPGA-negative phenotype of degQ gene knock-out mutant (ΔdegQ) was identified through whole genome sequencing. The mutation conferred an amino acid substitution of Ser103 to phenylalanine (S103F) in yabJ that belongs to the highly conserved YjgF/YER057c/UK114 family. Genetic experiments including LacZ-fusion assay of γPGA synthetic operon confirmed that the suppressor mutation (yabJS103F) was responsible for the recovery of γPGA production. The yabJ itself was not essential for the γPGA production and the mutant allele enabled γPGA production of the ΔdegQ strain even in the presence of wild type yabJ. Thus, yabJS103F was a dominant positive allele. degU-lacZ fusion gene was hyper-expressed in cells carrying the yabJS103F, but disruption of yabJ did not affect the transcription level of the degU-lacZ. These observations suggested that YabJ acquired a function to stimulate expression of degU by the S103F mutation which is involved in the regulation of γPGA synthesis.",
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AU - Hase, Sumitaka

AU - Shiwa, Yuh

AU - Yoshikawa, Hirofumi

AU - Sakakibara, Yasubumi

AU - Nguyen, Sy Le Thanh

AU - Kimura, Keitarou

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