Protein secretion in Bacillus brevis.
Many strains of Bacillus brevis were isolated from nature as very efficient producers of extracellular proteins. Strains identified as B. brevis including these protein-hyperproducers were reclassified into at least 6 species according to numerical analysis, DNA base composition, and DNA-DNA hybridization. We developed a host-vector system using appropriate strains of these Bacillus brevis as a host, which is excellent for the secretion of heterologous proteins. Utilizing the powerful promoters and signal peptide-coding regions of the cell wall protein gene, various expression-secretion vectors were constructed. The cell wall protein genes of these B. brevis are transcribed from multiple and tandemly arranged promoters. Transcription from P2, one of the major promoters among them, was enhanced at the early stationary phase of growth, when divalent cations in the medium was depleted and the cell wall protein layers started to be shed. Translation of the cell wall protein gene transcripts starts at the two sites located tandemly in the same reading frame. The two forms of secretory precursors, translation products from the two sites, are cleaved at the same position giving rise to the same mature proteins. The nucleotide sequence from the promoter to the translation start site is highly conserved in protein-hyperproducing B. brevis. For the efficient secretion of some heterologous proteins, protein-hypersecreting mutants had to be selected. The engineering of the signal peptide was also often necessary to obtain a good secretion of heterologous proteins.(ABSTRACT TRUNCATED AT 250 WORDS)
Udaka
S
S
Department of Applied Biological Sciences, Faculty of Agriculture, Nagoya University, Japan.
Yamagata
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001406:Bacillaceae / Q000145:classification / Q000235:genetics / Q000378:metabolism; D001407:Bacillus / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D017124:Conserved Sequence; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011994:Recombinant Proteins