1887

Abstract

An extracellular-protease-deficient mutant, ME142, was isolated from as a spontaneous erythromycin-resistant (Ery) clone. This mutant showed conditional sporulation and only sporulated normally in the absence of erythromycin. In the presence of the antibiotic, sporulation was greatly reduced. Production of extracellular proteases by ME142 also exhibited conditional deficiency, possibly due to pleiotropic effects of the sporulation deficiency. The production of protease was 2-10% that of the wild-type level in the presence of erythromycin. ME142 showed poor competence for transformation even in the absence of erythromycin; however, derivatives of ME142 were isolated which had the same Ery phenotype but which exhibited normal competence. One such mutant, ME162, was used as a host for the secretion of coli -lactamase. The amount of -lactamase in the culture supernatants of ME162 increased significantly when the cells were cultured with erythromycin, suggesting that proteolysis of the -lactamase in the supernatants of ME162 was greatly reduced as compared to that in the supernatants of the wild-type strain.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-135-4-777
1989-04-01
2021-10-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/135/4/mic-135-4-777.html?itemId=/content/journal/micro/10.1099/00221287-135-4-777&mimeType=html&fmt=ahah

References

  1. Chang S., Ho D., Gray O., Chang S.-Y., Mclaughlin J. 1983; Functional expression of human interferon genes and construction of a partition-proficient plasmid vector in B. subtilis . In Genetics of Industrial Microorganisms pp 227–231 Ikeda Y., Beppu T. Edited by Tokyo: Kodansha;
    [Google Scholar]
  2. Domoto T., Kobayashi K., Kobayashi Y. 1975; Erythromycin resistant, conditional asporogenous mutant of Bacillus subtilis . In Spores VI pp 307–313 Gerhardt P., Costilow R. N., Sadoff H. L. Edited by Washington, DC: American Society for Microbiology;
    [Google Scholar]
  3. Fahnestock S.R., Fischer K.E. 1986; Expression of the staphylococcal protein A gene in Bacillus subtilis by gene fusions utilizing the promoter from Bacillus amyloliquefaciens α-amylase gene. Journal of Bacteriology 165:796–804
    [Google Scholar]
  4. Fahnestock S.R., Fischer K.E. 1987; Protease-deficient Bacillus subtilis host strains from production of staphylococcal protein A. Applied and Environmental Microbiology 53:379–384
    [Google Scholar]
  5. Fujii M., Takagi M., Imanaka T., Aiba S. 1983; Molecular cloning of a thermostable neutral protease gene from Bacillus stearothermophilus in a vector plasmid and its expression in Bacillus stearothermophilus and Bacillus subtilis . Journal of Bacteriology 154:831–837
    [Google Scholar]
  6. Goldman R.C., Tipper D.J. 1979; Morphology and patterns of protein synthesis during sporulation of Bacillus subtilisEryrSpo(Ts) mutants. Journal of Bacteriology 138:625–637
    [Google Scholar]
  7. Honjo M., Akaoka A., Nakayama A., Shimada H., Furutani Y. 1985; Construction of the secretion vector containing the prepro-structure coding region of Bacillus amyloliquefaciens neutral protease gene and secretion of Bacillus subtilis a-amylase and human interferon-beta in Bacillus subtilis . Journal of Biotechnology 2:73–84
    [Google Scholar]
  8. Honjo M., Nakayama A., Iio A., Mita I., Kawamura K., Sawakura A., Furutani Y. 1987; Construction of a highly efficient host-vector system for secretion of heterologous protein in Bacillus subtilis . Journal of Biotechnology 6:191–204
    [Google Scholar]
  9. Iwakura M., Kawata M., Tsuda K., Tanaka T. 1988; Nucleotide sequence of the thymidylate synthase B and dihydrofolate reductase genes contained in one Bacillus subtilis operon. Gene 64:9–20
    [Google Scholar]
  10. Kamakura T., Kobayashi K., Tanaka T., Yamaguchi I., Endo T. 1987; Cloning and expression of a structural gene for blasticidin S deaminase, a nucleoside aminohydrolase. Agricultural and Biological Chemistry 51:3165–3168
    [Google Scholar]
  11. Kawamura F., Doi R.H. 1984; Construction of a Bacillus subtilis double mutant deficient in extracellular alkaline and neutral proteases. Journal of Bacteriology 160:442–444
    [Google Scholar]
  12. Leighton T.J., Doi R.H. 1971; The stability of messenger ribonucleic acid during sporulation in Bacillus subtilis . Journal of Biological Chemistry 246:3189–3195
    [Google Scholar]
  13. Mckenzie T., Hoshino T., Tanaka T., Sueoka N. 1986; The nucleotide sequence of pUB11O: some salient features in relation to replication and its regulation. Plasmid 15:93–103
    [Google Scholar]
  14. Nagami Y., Tanaka T. 1986; Molecular cloning and nucleotide sequence of a DNA fragment from Bacillus natto that enhances production of extracellular proteases and levansucrease in Bacillus subtilis . Journal of Bacteriology 166:20–28
    [Google Scholar]
  15. Nagami Y., Kimura M., Teranishi Y., Tanaka T. 1988; Construction of a new shuttle expression vector for Bacillus subtilis and Escherichia coli by using a polycistronic system. Gene 69:59–69
    [Google Scholar]
  16. Nakamura K., Furusato T., Shiroza T., Yamane K. 1985; Stable hyper-production of Escherichia coli /1-lactamase by Bacillus subtilis grown on a 0µ5 M succinate-medium using a B. subtilis α-amylase secretion vector. Biochemical and Biophysical Research Communications 128:601–606
    [Google Scholar]
  17. Neuhard J., Price A.R., Shack L., Thomassen E. 1978; Two thymidylate synthetases in Bacillus subtilis . Proceedings of the National Academy of Sciences of the United States of America 75:1194–1198
    [Google Scholar]
  18. Palva I., Sarvas M., Lehtovaara P., Sibakov M., Kääriäinen L. 1982; Secretion of Escherichia coli β-lactamase from Bacillus subtilis by the aid of α-amylase signal sequence. Proceedings of the National Academy of Sciences of the United States of America 79:5582–5586
    [Google Scholar]
  19. Palva I., Lehtovaara P., Kääriäinen L., Sibakov M., Cantnell K., Schein C.H., Kashiwagi K., Weissmann C. 1983; Secretion of interferon by B. subtilis . Gene 22:229–235
    [Google Scholar]
  20. Perego M., Hoch J.A. 1988; Sequence analysis and regulation of the hpr locus, a regulatory gene for protease production and sporulation in Bacillus subtilis . Journal of Bacteriology 170:2560–2567
    [Google Scholar]
  21. Piggot P.J., Coote J.G. 1976; Genetic aspects of bacterial endospore formation. Bacteriological Reviews 40:908–962
    [Google Scholar]
  22. Ruppen M., Band L., Henner D.J. 1986; Efficient expression of human growth hormone in Bacillus subtilis . In Bacillus Molecular Genetics and Biotechnology Applications, pp 423–432 Ganesan A. T., Hoch J. A. Edited by New York: Academic Press;
    [Google Scholar]
  23. Ruppen M.E., Alstine G.L., Van, Band L. 1988; Control of intracellular serine protease expression in Bacillus subtilis . Journal of Bacteriology 170:136–140
    [Google Scholar]
  24. Sadaie Y., Kada T. 1983; Formation of competent Bacillus subtilis cells. Journal of Bacteriology 153:813–821
    [Google Scholar]
  25. Sanders C.W., Schmidt B.J., Mallonee R.L., Guyer M.S. 1987; Secretion of human serum albumin from Bacillus subtilis . Journal of Bacteriology 169:2917–2925
    [Google Scholar]
  26. Schein C.H., Kashiwagi K., Fujisawa A., Weissmann C. 1986; Secretion of mature IFN-a2 and accumulation of uncleaved precursor by Bacillus subtilis transformed with a hybrid α-amylase signal sequence-IFN-α2 gene. Bio/technology 4:719–725
    [Google Scholar]
  27. Sharrock R.A., Leighton T., Wittmann H.G. 1981; Macrolide and aminoglycoside antibiotic resistance mutations in the Bacillus subtilis ribosome resulting in temperature-sensitive sporulation. Molecular and General Genetics 183:538–543
    [Google Scholar]
  28. Shiroza T., Nakazawa K., Tashiro N., Yamane K., Yanagi K., Yamazaki M., Tamura G., Saito H., Kawade K., Taniguchi T. 1985; Synthesis and secretion of biologically active mouse interferon-β using a Bacillus subtilis α-amylase secretion vector. Gene 34:1–8
    [Google Scholar]
  29. Spizizen J. 1958; Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribo- nucleate. Proceedings of the National Academy of Sciences of the United States of America 44:1072–1078
    [Google Scholar]
  30. Tanaka T., Kawano N. 1980; Cloning vehicles for the homologous Bacillus subtilis host-vector system. Gene 10:131–136
    [Google Scholar]
  31. Tanaka T., Kawata M., Nagami Y., Uchiyama H. 1987; prtR enhances the mRNA level of the B. subtilis extracellular proteases. Journal of Bacteriology 169:3044–3050
    [Google Scholar]
  32. Ulmanen I., Lundström K., Lethovaara P., Sarvas M., Ruohonen M., Palva I. 1985; Transcription and translation of foreign genes in Bacillus subtilis by the aid of a secretion vector. Journal of Bacteriology 162:176–182
    [Google Scholar]
  33. Uozumi T., Hoshino T., Miwa K., Horinouchi S., Beppu T., Arima K. 1977; Restriction and modification in Bacillus species. Genetic transformation of bacteria with DNA from different species, part 1. Molecular and General Genetics 152:65–69
    [Google Scholar]
  34. Wong S.-L., Kawamura F., Doi R.H. 1986; Use of the Bacillus subtilissubtilisin signal peptide for efficient secretion of TEM β-lactamase during growth. Journal of Bacteriology 168:1005–1009
    [Google Scholar]
  35. Yang M.Y., Ferrari E., Henner D.J. 1984; Cloning of the neutral protease gene of Bacillus subtilis and the use of the cloned gene to create an in vitro-derived mutation. Journal of Bacteriology 160:15–21
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-135-4-777
Loading
/content/journal/micro/10.1099/00221287-135-4-777
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error