Accumulation of an artificial cell wall-binding lipase by Bacillus subtilis wprA and/or sigD mutants.
A recombinant lipase, CWB-LipB, localized on the Bacillus subtilis cell surface and retaining lipase activity was unstable and not accumulated in a high yield. To improve the accumulation, we examined cell wall binding protease (wprA)- and/or sigma D (sigD)-deficient mutants, and also a NprE and AprA protease-deficient mutant as host strains. The nprE aprA mutation did not lead to a significant increase in the CWB-LipB accumulation. The wprA mutant accumulated a greater amount than the wild-type only in the stationary phase, but the sigD mutant accumulated a greater amount in both the exponential and stationary phases. The double mutant exhibited great accumulation of CWB-LipB, the amount being 36% of the total proteins extracted from the cell surface.
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Tokida, Ueda-shi, Nagano, Japan.
G
G
Kobayashi
J
J
Toida
T
T
Akamatsu
H
H
Yamamoto
T
T
Shida
J
J
Sekiguchi
D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins; D002473:Cell Wall / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005408:Flagellin / Q000235:genetics*; D008049:Lipase / Q000032:analysis / Q000378:metabolism*; D009154:Mutation; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000172:deficiency / Q000235:genetics*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D005408:Flagellin; C416532:flaB flagellin; D008049:Lipase; D012697:Serine Endopeptidases; C104015:WprA protein, Bacillus subtilis