Bacillus subtilis AprX involved in degradation of a heterologous protein during the late stationary growth phase

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 α-amylase-A522-PreS2 hybrid p...

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Bibliographic Details
Published in:Journal of bioscience and bioengineering 2007-08, Vol.104 (2), p.135-143
Main Authors: Kodama, Takeko, Endo, Keiji, Sawada, Kazuhisa, Ara, Katsutoshi, Ozaki, Katsuya, Kakeshita, Hiroshi, Yamane, Kunio, Sekiguchi, Junichi
Format: Article
Language:English
Subjects:
AprX protease
BACILLUS SUBTILIS
Bacillus subtilis - classification
Bacillus subtilis - enzymology
Bacillus subtilis - genetics
Bacillus subtilis - growth & development
Bacterial Proteins - metabolism
BIODEGRADACION
BIODEGRADATION
Biodegradation, Environmental
Biological and medical sciences
Biotechnology
Fundamental and applied biological sciences. Psychology
Hepatitis B virus
heterologous protein
MUTANT
MUTANTES
MUTANTS
PROTEASAS
PROTEASE
PROTEASES
PROTEINAS
PROTEINE
PROTEINS
Recombinant Proteins - metabolism
Serine Endopeptidases - metabolism
Species Specificity
zymography
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Summary: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 α-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 α-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 α-amylase-A522-PreS2 in the late stationary growth phase. Moreover, the production of α-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.
ISSN:1389-1723
1347-4421
DOI:10.1263/jbb.104.135