Isolation and characterization of Tn917-generated bacitracin deficient mutants of Bacillus licheniformis
Corresponding Author
Blagajana Herzog-Velikonja
Institute of Biology, University of Ljubljana, Ljubljana, Slovenia
*Corresponding author. Tel: 386 061 265 585; Fax: 386 61 273 390.Search for more papers by this authorZdravko Podlesek
Department of Biology, Biotechnical Faculty, Večna pot 111, University of Ljubljana, 61000 Ljubljana, Slovenia
Search for more papers by this authorMiklavž Grabnar
Department of Biology, Biotechnical Faculty, Večna pot 111, University of Ljubljana, 61000 Ljubljana, Slovenia
Search for more papers by this authorCorresponding Author
Blagajana Herzog-Velikonja
Institute of Biology, University of Ljubljana, Ljubljana, Slovenia
*Corresponding author. Tel: 386 061 265 585; Fax: 386 61 273 390.Search for more papers by this authorZdravko Podlesek
Department of Biology, Biotechnical Faculty, Večna pot 111, University of Ljubljana, 61000 Ljubljana, Slovenia
Search for more papers by this authorMiklavž Grabnar
Department of Biology, Biotechnical Faculty, Večna pot 111, University of Ljubljana, 61000 Ljubljana, Slovenia
Search for more papers by this authorAbstract
Abstract Two Tn917-generated bacitracin deficient mutants of Bacillus licheniformis were isolated. Southern blot analysis of chromosomal DNA extracted from both insertional mutants showed that Tn917 inserted in the vicinity of the gene coding for the enzyme BA2 of the bacitracin synthetase enzyme complex. Measurements of bacitracin synthetase activity in cell-free extracts and positive hybridization signals in the vicinity of the BA2 gene indicate that in both bacitracin deficient mutants Tn917 could be inserted in the BA1 gene or in segments involved in regulation. Thus, it could be possible that the genes for bacitracin synthetase are clustered in the B. licheniformis genome.
REFERENCES
- 1 Frøyshov, Ø. (1984) In Biotechnology of industrial antibiotics The bacitracins: properties, biosynthesis and fermentation and E.J. Vandamme, Ed) pp 665–694 Marcel Dekker Inc, New York .
- 2 Frøyshov, Ø. and Mathiesen, A. (1979) Tryptic cleavage of enzyme A in bacitracin synthetase FEBS Lett., 106, 275–278.
- 3
Korsnes, L.O.,
Gulliksen, M.,
Sundan, A. and
Nerland, A. (1986) Cloning of genes from Bacillus licheniformis involved in synthesis of the peptide antibiotic bacitracin
In
Bacillus molecular genetics and biotechnology applications
A.T. Ganesan and
J.A. Hoch, Eds) pp
283–294
Academic Press Inc,
Orlando
,
Florida
.
10.1016/B978-0-12-274155-5.50027-1 Google Scholar
- 4 Ishihara, H., Endo, Y., Abe, S. and Shimura, K. (1975) The presence of 4′-phosphopantheteine in the bacitracin synthetase FEBS Lett., 50, 43–46.
- 5 Marahiel, M.A., Krause, M. and Skarpeid, H.J. (1985) Cloning of the tyrocidine synthetase 1 gene from Bacillus brevis and its expression in Escherichia coli Mol. Gen. Genet., 201, 231–236.
- 6 Krause, M., Marahiel, M.A., Dohren, H. and Kleinkauf, H. (1985) Molecular cloning of an ornitine-activating fragment of the gramicidin S synthetase 2 gene from Bacillus brevis and its expression in Escherichia coli J. Bacteriol., 162, 1120–1125.
- 7 Ishihara, H., Hara, N. and Iwabuchi, T. (1989) Molecular cloning and expression in Escherichia coli of the Bacillus licheniformis bacitracin synthetase 2 gene J. Bacteriol., 171, 1705–1711.
- 8 Tomich, P.K., An, F.Y. and Clewell, D.B. (1980) Properties of erithromycin-inducible transposon Tn917 in Streptococcus faecalis J. Bacteriol., 141, 1366–1374.
- 9 Youngman, P.J. (1987) In Plasmids: a practical approach Plasmid vectors for recovering and exploiting Tn917 in Bacillus and other gram-positives and K. Hardy, Ed) pp 79–103 IRL Press, Oxford .
- 10 Ventra, L. and Weiss, A.S. (1989) Transposon-mediated restriction mapping of the Bacillus subtilis chromosome Gene, 78, 29–36.
- 11 Berg, D.E. and Howe, M.M. (1989) Mobile DNA American Society for Microbiology, Washington D.C .
- 12 Youngman, P.J., Perkins, J.B. and Losick, R. (1984) Construction of a cloning site near one end of Tn917 into which foreign DNA may be inserted without affecting transposition in Bacillus subtilis or expression of the transposon-born erm gene Plasmid, 12, 1–9.
- 13 Chang, S. and Cohen, S.N. (1979) High frequency transformation of Bacillus subtilis protoplast by plasmid DNA Mol. Gen. Genet., 168, 111–115.
- 14 Bourne, N. and Dancer, B.N. (1986) Regeneration of protoplasts of Bacillus subtilis 168 and closely related strains J. Gen. Microbiol., 132, 251–255.
- 15 Haavik, H.I. and Thomassen, S. (1973) A bacitracin-negative mutant of Bacillus licheniformis which is able to sporulate J. Gen. Microbiol., 76, 451–454.
- 16 Haavik, H.I. (1974) Studies on the formation of bacitracin by Bacillus licheniformis: effect of glucose J. Gen. Microbiol., 81, 383–390.
- 17 Shimura, K., Iwaki, M., Kanda, M., Hori, K., Kaji, E., Hasegawa, S. and Saito, Y. (1974) On the enzyme system obtained from some mutants of Bacillus brevis deficient in gramicidin S formation Biochim. Biophys. Acta, 338, 577–587.
- 18 Pitcher, D.G., Saunders, N.A. and Owen, R.J. (1989) Rapid extraction of bacterial genomic DNA with guanidium thiocyanate Lett. Appl. Microbiol., 8, 151–156.
- 19 Bron, S. (1990) Plasmids In Molecular biological methods for Bacillus C.R. Harwood and S.M. Cutting, Eds) pp 75–139 John Wiley and Sons Ltd, Chichester , England .
- 20 Maniatis, T., Fritsch, E. and Samobrook, J. (1982) Molecular cloning: a laboratory manual Cold Spring Harbor Laboratory, Cold Spring Harbor , New York .
- 21 Frøyshov, Ø. and Laland, S.G. (1974) On the biosynthesis of bacitracin by a soluble enzyme complex from Bacillus licheniformis Eur. J. Biochem., 46, 235–242.
- 22 Frøyshov, Ø. (1977) The production of bacitracin synthetase by Bacillus licheniformis ATCC 10716 FEBS Lett., 81, 315–318.
- 23 Mittenhuber, G., Weckermann, R. and Marahiel, M.A. (1989) Gene cluster containing the genes for tyrocidine synthetases 1 and 2 from Bacillus brevis: Evidence for an operon J. Bacteriol., 171, 4881–4887.
- 24 Krause, M. and Marahiel, M.A. (1988) Organization of the biosynthesis genes for the peptide antibiotic gramicidin S J. Bacteriol., 170, 4669–4674.
