Skip to main content
Log in

Identification of intracellular amylase activity in Streptococcus bovis and Streptococcus salivarius

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

The ruminal bacterium Streptococcus bovis has been demonstrated to produce an extracellular amylase activity. We previously reported on the cloning of a gene from S. bovis encoding for what was initially believed to be the extracellular amylase. DNA sequence analyses indicated that the amylase produced by the cloned gene did not match the N-terminus amino acid sequence of the purified extracellular amylase and contained no apparent leader sequence for secretion. Analyses of crude extracts demonstrated the presence of an intracellular amylase in S. bovis JB1 that differed in molecular weight (56,000) from that of the extracellular amylase (70,000). The 56,000 molecular weight amylase was identical to the amylase produced by Escherichia coli containing the cloned amylase gene. Low levels of intracellular amylase activity were also detected in other strains of S. bovis and also Streptococcus salivarius. Introduction of the plasmid pVA838 containing the cloned amylase gene into S. bovis and S. sanguis resulted in enhanced intracellular amylase production by both organisms. The amylase gene has been sequenced, and analysis of the deduced amino acid sequence for the amylase indicates a high degree of similarity with secreted amylases from Bacillus species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Literature Cited

  1. Allison MJ, Bucklin JA, Dougherty RW (1964) Ruminal changes after overfeeding with wheat and the effect of intraruminal inoculation on adaptation to a ration containing wheat. J Anim Sci 23:1164–1171

    Google Scholar 

  2. Awada A, Van der Auwera P, Meunier F, Daneau D, Klastersky J (1992) Streptococcal and enterococcal bacterimia in patients with cancer. Clin Infect Dis 15:33–48

    Google Scholar 

  3. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Google Scholar 

  4. Cotta MA, Whitehead TR (1993) Regulation and cloning of the gene encoding amylase activity of the ruminal bacterium Streptococcus bovis. Appl Environ Microbiol 59:189–196

    Google Scholar 

  5. Dubrow R, Edberg S, Wikfors E, Callan D, Troncale F, Vender R, Brand M, Yapp R (1991) Fecal carriage of Streptococcus bovis and colorectal adenomas. Gastroenterology 101:721–725

    Google Scholar 

  6. Farrow JAE, Kruze J, Phillips BA, Bramley AJ, Collins MD (1984) Taxonomic studies on Streptococcus bovis and Streptococcus equinus: description of Streptococcus alactolyticus sp. nov. and Streptococcus saccharolyticus sp. nov. Syst Appl Microbiol 5:468–482

    Google Scholar 

  7. Freer SN (1993) Purification and characterization of the extracellular α-amylase from Streptococcus bovis JB1. Appl Environ Microbiol 59:1398–1402

    Google Scholar 

  8. Hamilton IR (1976) Intracellular polysaccharide synthesis in cariogenic microorganism. In: Stile HM, Loesche WJ, O'Brien TL (eds) Proceedings in microbiology. Aspects of dental caries. Special supplement to Microbiology Abstracts, vol. 3. London: Intormation Retrieval, Inc., pp 638–701

    Google Scholar 

  9. Harley W (1992) Streptococcus bovis meningitis associated with a colonic villous adenoma. Clin Infect Dis 14:979–980

    Google Scholar 

  10. Hespell RB, Wolf R, Bothast RJ (1987) Fermentation of xylans by Butyrivibrio fibrisolvens and other ruminal bacteria. Appl Environ Microbiol 53:2849–2853

    Google Scholar 

  11. Janse BJH, Steyn AJC, Pretorius IS (1993) Regional sequence homologies in starch-degrading enzymes. Curr Genet 24:400–407

    Google Scholar 

  12. Klein RS, Recco RA, Catalano MT, Edberg SC, Casey JI, Steigbigel NH (1977) Association of Streptococcus bovis with carcinoma of the colon. N Engl J Med 297:800–802

    Google Scholar 

  13. Klein RS, Catalano MT, Edberg SC, Casey JI, Steigbigel NH (1979) Streptococcus bovis septicemia and carcinoma of the colon. Ann Intern Med 91:560–562

    Google Scholar 

  14. Lacks SA, Springhorn SS (1980) Renaturation of enzymes after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. J Biol Chem 255:7467–7473

    Google Scholar 

  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  16. Mackie RI, Gilchrist FMC (1978) Microbiological and chemical changes in the rumen during the stepwise adaptation of sheep to high concentrate diets. J Agr Sci 90:241–254

    Google Scholar 

  17. Mackie RI, Gilchrist FMC (1979) Changes in lactateproducing and lactate-utilizing bacteria in relation to pH in the rumen of sheep during stepwise adaptation to a highconcentrate diet. Appl Environ Microbiol 38:422–430

    Google Scholar 

  18. Macrina FL, Tobian JA, Jones KR, Evans RP, Clewell DB (1982) A cloning vector able to replicate in Escherichia coli and Streptococcus sanguis. Gene 19:345–353

    Google Scholar 

  19. Megran DW (1992) Enterococcal endocarditis. Clin Infect Dis 15:63–71

    Google Scholar 

  20. Pettersson G, Porath J (1966) A cellulolytic enzyme from Penicillium notatum. Methods Enzymol 8:603–607

    Google Scholar 

  21. Reynolds JG, Silva E, McCormack WM (1983) Association of Streptococcus bovis bacteremia with bowel disease. J Clin Microbiol 17:696–697

    Google Scholar 

  22. Russell JB, Baldwin RL (1979) Comparison of substrate affinities among several rumen bacteria: a possible determinant of rumen bacteria competition. Appl Environ Microbiol 37:531–536

    Google Scholar 

  23. Satoh E, Niimura Y, Uchimura T, Kozaki M, Komagata K (1993) Molecular cloning and expression of two α-amylase genes from Streptococcus bovis 148 in Escherichia coli. Appl Environ Microbiol 59:3669–3673

    Google Scholar 

  24. Slyter LL (1976) Influence of acidosis on rumen function. J Anim Sci 43:910–929

    Google Scholar 

  25. Walker GJ (1965) The cell-bound α-amylase of Streptococcus bovis. Biochem J 94:289–298

    Google Scholar 

  26. Whitehead TR, Cotta MA (1993) Development of a DNA probe for Streptococcus bovis by using a cloned amylase gene. J Clin Microbiol 31:2387–2391

    Google Scholar 

  27. Zarkin BA, Lillemoe KD, Cameron JL, Effron PN, Magnuson TH, Pitt HA (1990) The triad of Streptococcus bovis bactemia, colonic pathology, and liver disease. Ann Surg 211:786–792

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Whitehead, T.R., Cotta, M.A. Identification of intracellular amylase activity in Streptococcus bovis and Streptococcus salivarius . Current Microbiology 30, 143–148 (1995). https://doi.org/10.1007/BF00296199

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00296199

Keywords

Navigation