TY - JOUR
T1 - Bacillus subtilis AprX involved in degradation of a heterologous protein during the late stationary growth phase
AU - Kodama, Takeko
AU - Endo, Keiji
AU - Sawada, Kazuhisa
AU - Ara, Katsutoshi
AU - Ozaki, Katsuya
AU - Kakeshita, Hiroshi
AU - Yamane, Kunio
AU - Sekiguchi, Junichi
N1 - Funding Information:
This research was carried out as a part of the Project for the Development of a Technological Infrastructure for Industrial Bioprocesses through R and D on New Industrial Science and Technology Frontiers of the Ministry of Economy, Trade and Industry (METI), and supported by the New Energy and Industrial Technology Development Organization (NEDO). This research was also supported by Grants-in-Aid for Scientific Research on Priority Areas, Genome Biology (no. 12206005), the 21st Century COE Program, and Scientific Research (B) (no. 16380059) (J.S.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
PY - 2007/8
Y1 - 2007/8
N2 - 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.
AB - 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.
KW - AprX protease
KW - Bacillus subtilis
KW - heterologous protein
KW - zymography
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U2 - 10.1263/jbb.104.135
DO - 10.1263/jbb.104.135
M3 - Article
C2 - 17884659
AN - SCOPUS:34548604939
SN - 1389-1723
VL - 104
SP - 135
EP - 143
JO - Journal of Bioscience and Bioengineering
JF - Journal of Bioscience and Bioengineering
IS - 2
ER -
