Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1990 May;56(5):1221–1228. doi: 10.1128/aem.56.5.1221-1228.1990

Cellulose digestion and cellulase regulation and distribution in Fibrobacter succinogenes subsp. succinogenes S85.

L Huang 1, C W Forsberg 1
PMCID: PMC184386  PMID: 2339881

Abstract

Fibrobacter succinogenes subsp. succinogenes S85 initiated growth on microcrystalline cellulose without a lag whether inoculated from a glucose, cellobiose, or cellulose culture. During growth on cellulose, there was no accumulation of soluble carbohydrate. When the growth medium contained either glucose or cellobiose in combination with microcrystalline cellulose, there was a lag in cellulose digestion until all of the soluble sugar had been utilized, suggesting an end product feedback mechanism that affects cellulose digestion. Cl-stimulated cellobiosidase and periplasmic cellodextrinase were produced under all growth conditions tested, indicating constitutive synthesis. Both cellobiosidases were cell associated until the stationary phase of growth, whereas proteins antigenically related to the Cl-stimulated cellobiosidase and a proportion of the endoglucanase were released into the extracellular culture fluid during growth, irrespective of the substrate. Immunoelectron microscopy of cells with a polyclonal antibody to Cl-stimulated cellobiosidase as the primary antibody and 10-nm-diameter gold particles conjugated to goat anti-rabbit antibodies as the second antibody revealed protrusions of the outer surface which were selectively labeled with gold, suggesting that Cl-stimulated cellobiosidase was located on the protrusions. These data support the contention that the protrusions have a role in cellulose hydrolysis; however, this interpretation is complicated by reactivity of the antibodies with a large number of other proteins that possess related antigenic epitopes.

Full text

PDF
1225

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Forsberg C. W., Beveridge T. J., Hellstrom A. Cellulase and Xylanase Release from Bacteroides succinogenes and Its Importance in the Rumen Environment. Appl Environ Microbiol. 1981 Nov;42(5):886–896. doi: 10.1128/aem.42.5.886-896.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ghangas G. S., Wilson D. B. Cloning of the Thermomonospora fusca Endoglucanase E2 Gene in Streptomyces lividans: Affinity Purification and Functional Domains of the Cloned Gene Product. Appl Environ Microbiol. 1988 Oct;54(10):2521–2526. doi: 10.1128/aem.54.10.2521-2526.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gong J., Forsberg C. W. Factors affecting adhesion of Fibrobacter succinogenes subsp. succinogenes S85 and adherence-defective mutants to cellulose. Appl Environ Microbiol. 1989 Dec;55(12):3039–3044. doi: 10.1128/aem.55.12.3039-3044.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Groleau D., Forsberg C. W. Cellulolytic activity of the rumen bacterium Bacteroides succinogenes. Can J Microbiol. 1981 May;27(5):517–530. doi: 10.1139/m81-077. [DOI] [PubMed] [Google Scholar]
  6. HALLIWELL G., BRYANT M. P. THE CELLULOLYTIC ACTIVITY OF PURE STRAINS OF BACTERIA FROM THE RUMEN OF CATTLE. J Gen Microbiol. 1963 Sep;32:441–448. doi: 10.1099/00221287-32-3-441. [DOI] [PubMed] [Google Scholar]
  7. Hiltner P., Dehority B. A. Effect of soluble carbohydrates on digestion of cellulose by pure cultures of rumen bacteria. Appl Environ Microbiol. 1983 Sep;46(3):642–648. doi: 10.1128/aem.46.3.642-648.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Huang L., Forsberg C. W. Isolation of a Cellodextrinase from Bacteroides succinogenes. Appl Environ Microbiol. 1987 May;53(5):1034–1041. doi: 10.1128/aem.53.5.1034-1041.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Huang L., Forsberg C. W. Purification and Comparison of the Periplasmic and Extracellular Forms of the Cellodextrinase from Bacteroides succinogenes. Appl Environ Microbiol. 1988 Jun;54(6):1488–1493. doi: 10.1128/aem.54.6.1488-1493.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Huang L., Forsberg C. W., Thomas D. Y. Purification and characterization of a chloride-stimulated cellobiosidase from Bacteroides succinogenes S85. J Bacteriol. 1988 Jul;170(7):2923–2932. doi: 10.1128/jb.170.7.2923-2932.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huang L., McGavin M., Forsberg C. W., Lam J. S., Cheng K. J. Antigenic nature of the chloride-stimulated cellobiosidase and other cellulases of Fibrobacter succinogenes subsp. succinogenes S85 and related fresh isolates. Appl Environ Microbiol. 1990 May;56(5):1229–1234. doi: 10.1128/aem.56.5.1229-1234.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Johnson E. A., Sakajoh M., Halliwell G., Madia A., Demain A. L. Saccharification of Complex Cellulosic Substrates by the Cellulase System from Clostridium thermocellum. Appl Environ Microbiol. 1982 May;43(5):1125–1132. doi: 10.1128/aem.43.5.1125-1132.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lam J. S., Lam M. Y., MacDonald L. A., Hancock R. E. Visualization of Pseudomonas aeruginosa O antigens by using a protein A-dextran-colloidal gold conjugate with both immunoglobulin G and immunoglobulin M monoclonal antibodies. J Bacteriol. 1987 Aug;169(8):3531–3538. doi: 10.1128/jb.169.8.3531-3538.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lamed R., Naimark J., Morgenstern E., Bayer E. A. Specialized cell surface structures in cellulolytic bacteria. J Bacteriol. 1987 Aug;169(8):3792–3800. doi: 10.1128/jb.169.8.3792-3800.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McGavin M. J., Forsberg C. W., Crosby B., Bell A. W., Dignard D., Thomas D. Y. Structure of the cel-3 gene from Fibrobacter succinogenes S85 and characteristics of the encoded gene product, endoglucanase 3. J Bacteriol. 1989 Oct;171(10):5587–5595. doi: 10.1128/jb.171.10.5587-5595.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McGavin M., Forsberg C. W. Isolation and characterization of endoglucanases 1 and 2 from Bacteroides succinogenes S85. J Bacteriol. 1988 Jul;170(7):2914–2922. doi: 10.1128/jb.170.7.2914-2922.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McGavin M., Lam J., Forsberg C. W. Regulation and distribution of Fibrobacter succinogenes subsp. succinogenes S85 endoglucanases. Appl Environ Microbiol. 1990 May;56(5):1235–1244. doi: 10.1128/aem.56.5.1235-1244.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. SCOTT H. W., DEHORITY B. A. VITAMIN REQUIREMENTS OF SEVERAL CELLULOLYTIC RUMEN BACTERIA. J Bacteriol. 1965 May;89:1169–1175. doi: 10.1128/jb.89.5.1169-1175.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smith W. R., Yu I., Hungate R. E. Factors affecting cellulolysis by Ruminococcus albus. J Bacteriol. 1973 May;114(2):729–737. doi: 10.1128/jb.114.2.729-737.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Updegraff D. M. Semimicro determination of cellulose in biological materials. Anal Biochem. 1969 Dec;32(3):420–424. doi: 10.1016/s0003-2697(69)80009-6. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES