Europe PMC

This website requires cookies, and the limited processing of your personal data in order to function. By using the site you are agreeing to this as outlined in our privacy notice and cookie policy.

Biochemical evidence that starch breakdown by Bacteroides thetaiotaomicron involves outer membrane starch-binding sites and periplasmic starch-degrading enzymes.

Author information

Affiliations

  • 1. Department of Microbiology, University of Illinois, Urbana 61801.
      Authors
    • Anderson KL 1
    (1 author)

Journal of Bacteriology, 01 Jun 1989, 171(6):3192-3198
https://doi.org/10.1128/jb.171.6.3192-3198.1989 PMID: 2722747 PMCID: PMC210036

Free full text in Europe PMC

Abstract 

Bacteroides thetaiotaomicron can utilize amylose, amylopectin, and pullulan as sole sources of carbon and energy. The enzymes that degrade these polysaccharides were found to be primarily cell associated rather than extracellular. Although some activity was detected in extracellular fluid, this appeared to be the result of cell lysis. The cell-associated amylase, amylopectinase, and pullulanase activities partitioned similarly to the periplasmic marker, acid phosphatase, when cells were exposed to a cold-shock treatment. Two other enzymes associated with starch breakdown, alpha-glucosidase and maltase, appeared to be located in the cytoplasm. Intact cells of B. thetaiotaomicron were found to bind 14C-starch. Binding was probably mediated by a protein because it was saturable and was decreased by treatment of cells with proteinase K. Results of competition experiments showed that the starch-binding proteins had a preference for maltodextrins larger than maltohexaose and a low affinity for maltose and maltotriose. Both the degradative enzymes and starch binding were induced by maltose. These findings indicate that starch utilization by B. thetaiotaomicron apparently does not involve secretion of extracellular enzymes. Rather, binding of the starch molecule to the cell surface appears to be a first step to passing the molecule through the outer membrane and into the periplasmic space.

Free full text 

Logo of jbacterLink to Publisher's site
J Bacteriol. 1989 Jun; 171(6): 3192–3198.
PMCID: PMC210036
PMID: 2722747

Biochemical evidence that starch breakdown by Bacteroides thetaiotaomicron involves outer membrane starch-binding sites and periplasmic starch-degrading enzymes.

K L Anderson and A A Salyers
Author information Copyright and License information Disclaimer

Abstract

Bacteroides thetaiotaomicron can utilize amylose, amylopectin, and pullulan as sole sources of carbon and energy. The enzymes that degrade these polysaccharides were found to be primarily cell associated rather than extracellular. Although some activity was detected in extracellular fluid, this appeared to be the result of cell lysis. The cell-associated amylase, amylopectinase, and pullulanase activities partitioned similarly to the periplasmic marker, acid phosphatase, when cells were exposed to a cold-shock treatment. Two other enzymes associated with starch breakdown, alpha-glucosidase and maltase, appeared to be located in the cytoplasm. Intact cells of B. thetaiotaomicron were found to bind 14C-starch. Binding was probably mediated by a protein because it was saturable and was decreased by treatment of cells with proteinase K. Results of competition experiments showed that the starch-binding proteins had a preference for maltodextrins larger than maltohexaose and a low affinity for maltose and maltotriose. Both the degradative enzymes and starch binding were induced by maltose. These findings indicate that starch utilization by B. thetaiotaomicron apparently does not involve secretion of extracellular enzymes. Rather, binding of the starch molecule to the cell surface appears to be a first step to passing the molecule through the outer membrane and into the periplasmic space.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.3M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
 
icon of scanned page 3192
3192
icon of scanned page 3193
3193
icon of scanned page 3194
3194
icon of scanned page 3195
3195
icon of scanned page 3196
3196
icon of scanned page 3197
3197
icon of scanned page 3198
3198
 

Images in this article

Image
Image
on p.3195
Image
Image
on p.3196
Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Anderson KL, Salyers AA. Genetic evidence that outer membrane binding of starch is required for starch utilization by Bacteroides thetaiotaomicron. J Bacteriol. 1989 Jun;171(6):3199–3204. [Europe PMC free article] [Abstract] [Google Scholar]
  • Beacham IR. Periplasmic enzymes in gram-negative bacteria. Int J Biochem. 1979;10(11):877–883. [Abstract] [Google Scholar]
  • Dygert S, Li LH, Florida D, Thoma JA. Determination of reducing sugar with improved precision. Anal Biochem. 1965 Dec;13(3):367–374. [Abstract] [Google Scholar]
  • Ford JR, Nunley JA, 2nd, Li YT, Chambers RP, Cohen W. A continuously monitored spectrophotometric assay of glycosidases with nitrophenyl glycosides. Anal Biochem. 1973 Jul;54(1):120–128. [Abstract] [Google Scholar]
  • Gherardini F, Babcock M, Salyers AA. Purification and characterization of two alpha-galactosidases associated with catabolism of guar gum and other alpha-galactosides by Bacteroides ovatus. J Bacteriol. 1985 Feb;161(2):500–506. [Europe PMC free article] [Abstract] [Google Scholar]
  • Huang L, Forsberg CW. Isolation of a Cellodextrinase from Bacteroides succinogenes. Appl Environ Microbiol. 1987 May;53(5):1034–1041. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kasahara M, Anraku Y. Succinate dehydrogenase of Escherichia coli membrane vesicles. Activation and properties of the enzyme. J Biochem. 1974 Nov;76(5):959–966. [Abstract] [Google Scholar]
  • Kotarski SF, Linz J, Braun DM, Salyers AA. Analysis of outer membrane proteins which are associated with growth of Bacteroides thetaiotaomicron on chondroitin sulfate. J Bacteriol. 1985 Sep;163(3):1080–1086. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kotarski SF, Salyers AA. Isolation and characterization of outer membranes of Bacteroides thetaiotaomicron grown on different carbohydrates. J Bacteriol. 1984 Apr;158(1):102–109. [Europe PMC free article] [Abstract] [Google Scholar]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [Abstract] [Google Scholar]
  • Macy JM, Ljungdahl LG, Gottschalk G. Pathway of succinate and propionate formation in Bacteroides fragilis. J Bacteriol. 1978 Apr;134(1):84–91. [Europe PMC free article] [Abstract] [Google Scholar]
  • MALAMY M, HORECKER BL. The localization of alkaline phosphatase in E. coli K12. Biochem Biophys Res Commun. 1961 Jun 2;5:104–108. [Abstract] [Google Scholar]
  • McCarthy RE, Kotarski SF, Salyers AA. Location and characteristics of enzymes involved in the breakdown of polygalacturonic acid by Bacteroides thetaiotaomicron. J Bacteriol. 1985 Feb;161(2):493–499. [Europe PMC free article] [Abstract] [Google Scholar]
  • Moore WE, Holdeman LV. Human fecal flora: the normal flora of 20 Japanese-Hawaiians. Appl Microbiol. 1974 May;27(5):961–979. [Europe PMC free article] [Abstract] [Google Scholar]
  • Salyers AA, O'Brien M. Cellular location of enzymes involved in chondroitin sulfate breakdown by Bacteroides thetaiotaomicron. J Bacteriol. 1980 Aug;143(2):772–780. [Europe PMC free article] [Abstract] [Google Scholar]
  • Smith KA, Salyers AA. Cell-associated pullulanase from Bacteroides thetaiotaomicron: cloning, characterization, and insertional mutagenesis to determine role in pullulan utilization. J Bacteriol. 1989 Apr;171(4):2116–2123. [Europe PMC free article] [Abstract] [Google Scholar]
Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

Full text links 

Read article at publisher's site: https://doi.org/10.1128/jb.171.6.3192-3198.1989

Read article for free, from open access legal sources, via Unpaywall: https://europepmc.org/articles/pmc210036?pdf=renderUnpaywall

Free to read at jb.asm.org
http://jb.asm.org/cgi/content/abstract/171/6/3192

Free after 4 months at jb.asm.org
http://jb.asm.org/cgi/reprint/171/6/3192

Citations & impact 

Impact metrics

83
Citations
Jump to Citations

Citations of article over time

Article citations

Go to all (83) article citations

Similar Articles 

To arrive at the top five similar articles we use a word-weighted algorithm to compare words from the Title and Abstract of each citation.

Funding 

Funders who supported this work.

NIAID NIH HHS (1)

NIDDK NIH HHS (1)