Overproduction of a dextranase inhibitor by Streptococcus sobrinus mutants

S. Y. Wanda, A. Camilli, H. M. Murchison, R. Curtiss

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

An inhibitor of Streptococcus sobrinus endodextranase was detected in the extracellular fractions of UAB66 mutants identified following ethyl methanesulfonate mutagenesis as either devoid of dextranase activity (Dex-) or overproducing water-soluble glucan. The two groups of mutants had the same phenotype and displayed no dextranase activity in assays of extracellular fractions (H. Murchison, S. Larrimore, and R. Curtiss III, Infect. Immun. 34:1044-1055, 1981) and had been shown to be defective in adherence (Adh-) and capable of inhibiting adherence of wild-type strains during cocultivation in vitro (H. Murchison, S. Larrimore, and R. Curtiss III, Infect. Immun. 50:826-832, 1985) and in vivo in gnotobiotic rats (K. Takada, T. Shiota, R. Curtiss III, and S. M. Michalek, Infect. Immun. 50:833-843, 1985). By analysis of proteins in Western blots (immunoblots) and following blue dextran-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (BD-SDS- PAGE), it was demonstrated that these Dex- mutants did synthesize enzymatically active dextranase. From the results of mixing experiments, it was determined that these Dex- Adh- mutants produced enhanced amounts of a cell surface-localized or a cell-associated dextranase inhibitor (Dei). Dei was heat stable but trypsin sensitive. By adding excess dextranase following BD-SDS-PAGE, Dei was detected as blue bands with apparent molecular masses of 43, 40, 37, 27, and 23 kDa. Dei competitively inhibits dextranase activity and is synthesized by wild-type S. sobrinus strains, with the amount varying depending upon growth medium and stage in the growth cycle. R. M. Hamelik and M. M. McCabe (Biochem. Biophys. Res. Commun. 106:875-880, 1982) previously described a Dei in a wild-type S. sobrinus strain.

Original languageEnglish (US)
Pages (from-to)7206-7212
Number of pages7
JournalJournal of Bacteriology
Volume176
Issue number23
StatePublished - 1994
Externally publishedYes

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Dextranase
Streptococcus sobrinus
Polyacrylamide Gel Electrophoresis
Western Blotting
Germ-Free Life
Ethyl Methanesulfonate
Glucans
Growth
Coculture Techniques
Mutagenesis
Sodium Dodecyl Sulfate
Trypsin

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Wanda, S. Y., Camilli, A., Murchison, H. M., & Curtiss, R. (1994). Overproduction of a dextranase inhibitor by Streptococcus sobrinus mutants. Journal of Bacteriology, 176(23), 7206-7212.

Overproduction of a dextranase inhibitor by Streptococcus sobrinus mutants. / Wanda, S. Y.; Camilli, A.; Murchison, H. M.; Curtiss, R.

In: Journal of Bacteriology, Vol. 176, No. 23, 1994, p. 7206-7212.

Research output: Contribution to journalArticle

Wanda, SY, Camilli, A, Murchison, HM & Curtiss, R 1994, 'Overproduction of a dextranase inhibitor by Streptococcus sobrinus mutants', Journal of Bacteriology, vol. 176, no. 23, pp. 7206-7212.
Wanda SY, Camilli A, Murchison HM, Curtiss R. Overproduction of a dextranase inhibitor by Streptococcus sobrinus mutants. Journal of Bacteriology. 1994;176(23):7206-7212.
Wanda, S. Y. ; Camilli, A. ; Murchison, H. M. ; Curtiss, R. / Overproduction of a dextranase inhibitor by Streptococcus sobrinus mutants. In: Journal of Bacteriology. 1994 ; Vol. 176, No. 23. pp. 7206-7212.
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