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

Takeko Kodama, Keiji Endo, Kazuhisa Sawada, Katsutoshi Ara, Katsuya Ozaki, Hiroshi Kakeshita, Kunio Yamane, Junichi Sekiguchi

Research output: Contribution to journalArticle

13 Citations (Scopus)

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 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.

Original languageEnglish
Pages (from-to)135-143
Number of pages9
JournalJournal of Bioscience and Bioengineering
Volume104
Issue number2
DOIs
Publication statusPublished - 2007 Aug 1

Keywords

  • AprX protease
  • Bacillus subtilis
  • heterologous protein
  • zymography

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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