Effect of Bacillus subtilis spo0A mutation on cell wall lytic enzymes and extracellular proteases, and prevention of cell lysis.
The Bacillus subtilis spo0A mutant is an adequate host for extracellular protein production (e.g., alpha-amylase). However the mutant was prone to cell lysis. SDS-PAGE and zymography of cell wall lytic proteins indicated that the spo0A mutant contained high amounts of two major autolysins (LytC [CwlB] and LytD [CwlG]) and two minor cell wall lytic enzymes (LytE [CwlF] and LytF [CwlE]). On the other hand, the expression of eight extracellular protease genes was very poor or absent in the spo0A mutant. An eight-extracellular-protease-deficient mutant (Dpr8 strain) was constructed and the strain also exhibited cell lysis. The autolysins from the spo0A mutant were degraded by the supernatant of the wild type but not degraded by that of the Dpr8 mutant. These results suggest that the extensive cell lysis of the spo0A mutant was partially caused by the stability of autolysins via the decrease of the extracellular proteases. The introduction of a major autolysin and/or SigD mutations into the spo0A mutant was effective for preventing cell lysis.
Kodama
Takeko
T
Tochigi Research Laboratories of Kao Corporation, 2606 Akabane, Ichikai, Haga, Tochigi 321-3497, Japan.
Endo
Keiji
K
Ara
Katsutoshi
K
Ozaki
Katsuya
K
Kakeshita
Hiroshi
H
Yamane
Kunio
K
Sekiguchi
Junichi
J
D001426:Bacterial Proteins / Q000235:genetics*; D001433:Bacteriolysis / Q000502:physiology*; D002462:Cell Membrane / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000166:cytology* / Q000502:physiology*; D024861:Genetic Enhancement / Q000379:methods*; D009154:Mutation; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics*
D001426:Bacterial Proteins; C080950:Spo0A protein, Bacillus subtilis; D014157:Transcription Factors; D010447:Peptide Hydrolases; D009238:N-Acetylmuramoyl-L-alanine Amidase