The effects of ftsZ mutation on the production of recombinant protein in Bacillus subtilis.
In this paper, the possibility of using a mutation of ftsZ as a pseudo-spore mutant is investigated. ftsZ, which is essential for cell division and sporulation of Bacillus subtilis, was placed under the spac promoter, which is inducible with isopropyl thiogalactose (IPTG). Cell growth of the ftsZ mutant and its beta-galactosidase activity under the aprE promoter were compared with the wild type. In the presence of 1 mM IPTG, cell growth of the ftsZ mutant was almost the same as that of the wild type and its sporulation frequency was slightly lower than that of the wild type. However, under uninduced conditions, cell growth of ftsZ mutant was severely impaired. When induced with 0.2 mM IPTG, the ftsZ mutant showed about 13 times higher beta-galactosidase activity than the wild type. When the ftsZ mutant was used for secretory production of subtilisin, only three times higher extracellular subtilisin activity was measured, compared with the wild type. By real-time PCR investigation, it was revealed that the ftsZ mutant intracellular mRNA level for subtilisin was more than 16 times higher, compared with the wild type. However, it appears that the secretion pathway is somewhat damaged in the ftsZ mutant. These results suggest that the cell division mutant can also be used like a sporulation mutant to produce recombinant proteins, with a precise control of cell growth and induction.
Park
In-Suk
IS
School of Chemical Engineering and Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea.
Kim
June-Hyung
JH
Kim
Byung-Gee
BG
D001412:Bacillus subtilis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial; D007544:Isopropyl Thiogalactoside; D026901:Membrane Transport Proteins / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D021381:Protein Transport; D012329:RNA, Bacterial / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis*; D011995:Recombination, Genetic; D020860:Subtilisin / Q000378:metabolism; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000032:analysis
C088074:AprE protein, Bacteria; D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D026901:Membrane Transport Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside; D001616:beta-Galactosidase; D020860:Subtilisin