Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Dec;164(3):1182–1187. doi: 10.1128/jb.164.3.1182-1187.1985

Efficient synthesis and secretion of a thermophilic alpha-amylase by protein-producing Bacillus brevis 47 carrying the Bacillus stearothermophilus amylase gene.

N Tsukagoshi, S Iritani, T Sasaki, T Takemura, H Ihara, Y Idota, H Yamagata, S Udaka
PMCID: PMC219313  PMID: 2999073

Abstract

Bacillus subtilis and Bacillus brevis 47-5, carrying the Bacillus stearothermophilus alpha-amylase gene on pUB110 (pBAM101), synthesized the same alpha-amylase as the donor strain as determined by the enzyme's thermal stability and NH2-terminal amino acid sequence. Regardless of the host, the 34-amino acid signal peptide of the enzyme was processed at exactly the same site between two alanine residues. B. brevis 47-5(pBAM101) secreted the enzyme most efficiently of the hosts examined, 100, 15, and 5 times more than B. stearothermophilus, Escherichia coli HB101(pH1301), and B. subtilis 1A289(pBAM101), respectively. The efficient secretion of the enzyme in B. brevis 47-5(pBAM101) was suggested to be due to the unique properties of the cell wall of this organism.

Full text

PDF
1182

Images in this article

1183Image
on p.1183
1184Image
on p.1184

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  3. Ihara H., Sasaki T., Tsuboi A., Yamagata H., Tsukagoshi N., Udaka S. Complete nucleotide sequence of a thermophilic alpha-amylase gene: homology between prokaryotic and eukaryotic alpha-amylases at the active sites. J Biochem. 1985 Jul;98(1):95–103. doi: 10.1093/oxfordjournals.jbchem.a135279. [DOI] [PubMed] [Google Scholar]
  4. Inouye M., Halegoua S. Secretion and membrane localization of proteins in Escherichia coli. CRC Crit Rev Biochem. 1980;7(4):339–371. doi: 10.3109/10409238009105465. [DOI] [PubMed] [Google Scholar]
  5. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Mézes P. S., Blacher R. W., Lampen J. O. Processing of Bacillus cereus 569/H beta-lactamase I in Escherichia coli and Bacillus subtilis. J Biol Chem. 1985 Jan 25;260(2):1218–1223. [PubMed] [Google Scholar]
  8. Ohmizu H., Sasaki T., Tsukagoshi N., Udaka S., Kaneda N., Yagi K. Major proteins released by a protein-producing bacterium, Bacillus brevis 47, are derived from cell wall protein. J Biochem. 1983 Oct;94(4):1077–1084. doi: 10.1093/oxfordjournals.jbchem.a134450. [DOI] [PubMed] [Google Scholar]
  9. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  10. Stephens M. A., Ortlepp S. A., Ollington J. F., McConnell D. J. Nucleotide sequence of the 5' region of the Bacillus licheniformis alpha-amylase gene: comparison with the B. amyloliquefaciens gene. J Bacteriol. 1984 Apr;158(1):369–372. doi: 10.1128/jb.158.1.369-372.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Takahashi W., Yamagata H., Yamaguchi K., Tsukagoshi N., Udaka S. Genetic transformation of Bacillus brevis 47, a protein-secreting bacterium, by plasmid DNA. J Bacteriol. 1983 Dec;156(3):1130–1134. doi: 10.1128/jb.156.3.1130-1134.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Takkinen K., Pettersson R. F., Kalkkinen N., Palva I., Söderlund H., Käriäinen L. Amino acid sequence of alpha-amylase from Bacillus amyloliquefaciens deduced from the nucleotide sequence of the cloned gene. J Biol Chem. 1983 Jan 25;258(2):1007–1013. [PubMed] [Google Scholar]
  13. Tsuboi A., Tsukagoshi N., Udaka S. Reassembly in vitro of hexagonal surface arrays in a protein-producing bacterium, Bacillus brevis 47. J Bacteriol. 1982 Sep;151(3):1485–1497. doi: 10.1128/jb.151.3.1485-1497.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tsukagoshi N., Ihara H., Yamagata H., Udaka S. Cloning and expression of a thermophilic alpha-amylase gene from Bacillus stearothermophilus in Escherichia coli. Mol Gen Genet. 1984;193(1):58–63. doi: 10.1007/BF00327414. [DOI] [PubMed] [Google Scholar]
  15. Tsukagoshi N., Tabata R., Takemura T., Yamagata H., Udaka S. Molecular cloning of a major cell wall protein gene from protein-producing Bacillus brevis 47 and its expression in Escherichia coli and Bacillus subtilis. J Bacteriol. 1984 Jun;158(3):1054–1060. doi: 10.1128/jb.158.3.1054-1060.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ulmanen I., Lundström K., Lehtovaara P., Sarvas M., Ruohonen M., Palva I. Transcription and translation of foreign genes in Bacillus subtilis by the aid of a secretion vector. J Bacteriol. 1985 Apr;162(1):176–182. doi: 10.1128/jb.162.1.176-182.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Yamada H., Tsukagoshi N., Udaka S. Morphological alterations of cell wall concomitant with protein release in a protein-producing bacterium, Bacillus brevis 47. J Bacteriol. 1981 Oct;148(1):322–332. doi: 10.1128/jb.148.1.322-332.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yamazaki H., Ohmura K., Nakayama A., Takeichi Y., Otozai K., Yamasaki M., Tamura G., Yamane K. Alpha-amylase genes (amyR2 and amyE+) from an alpha-amylase-hyperproducing Bacillus subtilis strain: molecular cloning and nucleotide sequences. J Bacteriol. 1983 Oct;156(1):327–337. doi: 10.1128/jb.156.1.327-337.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yang M., Galizzi A., Henner D. Nucleotide sequence of the amylase gene from Bacillus subtilis. Nucleic Acids Res. 1983 Jan 25;11(2):237–249. doi: 10.1093/nar/11.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES