title: Bacillus subtilis AprX involved in degradation of a heterologous protein during the late stationary growth phase.
abstract: In Bacillus subtilis, extracellular protease-deficient mutants have been used in attempts to increase the productivity of heterologous proteins. We detected protease activity of AprX using protease zymography in the culture medium at the late stationary growth phase. An alpha-amylase-A522-PreS2 hybrid protein, in which the PreS2 antigen of human hepatitis B virus (HBV) is fused with the N-terminal 522-amino-acid polypeptide of B. subtilis alpha-amylase, has been produced in multiple-protease-deficient mutants. The B. subtilis KA8AX strain, which is deficient in eight extracellular proteases and AprX, did not show the proteolysis of alpha-amylase-A522-PreS2 in the late stationary growth phase. Moreover, the production of alpha-amylase-A522-PreS2 was about 80 mg/l, which was eight times higher than that by the KA8AX strain previously reported. In addition, we showed the degradation of the heterologous protein by AprX that leaked to the culture medium (probably caused by cell lysis) during the late stationary growth phase.
authors: Biological Science Laboratories, Kao Corporation, 2606 Akabane, Ichikai, Haga, Tochigi 321-3497, Japan.
authors: Takeko
authors: T
authors: Kodama
authors: Keiji
authors: K
authors: Endo
authors: Kazuhisa
authors: K
authors: Sawada
authors: Katsutoshi
authors: K
authors: Ara
authors: Katsuya
authors: K
authors: Ozaki
authors: Hiroshi
authors: H
authors: Kakeshita
authors: Kunio
authors: K
authors: Yamane
authors: Junichi
authors: J
authors: Sekiguchi
mesh_terms: D001412:Bacillus subtilis / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D001426:Bacterial Proteins / Q000378:metabolism*; D001673:Biodegradation, Environmental; D011994:Recombinant Proteins / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; C402704:AprX protein, bacteria; D012697:Serine Endopeptidases