Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1987 Feb;53(2):304–308. doi: 10.1128/aem.53.2.304-308.1987

Factors Involved in Hydrolysis of Microcrystalline Cellulose by Acetivibrio cellulolyticus†

C Roger MacKenzie 1,*, Girishchandra B Patel 1, Doris Bilous 1
PMCID: PMC203656  PMID: 16347279

Abstract

Acetivibrio cellulolyticus cellulase obtained by the water elution of residual cellulose from the growth medium was compared with the cellulase activity present in culture supernatants. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that water elution released most of the protein bands which adhered to undigested cellulose from the culture medium. The enzyme in the culture supernatant and that eluted from residual cellulose had specific activities for Avicel hydrolysis that were 20- to 40-fold greater than that of Trichoderma reesei cellulase. However, Ca2+ and a reducing agent such as dithiothreitol were required for maximum Avicel hydrolysis rates by these A. cellulolyticus enzyme preparations. The effect of these agents on p-nitrophenyl lactopyranoside hydrolysis suggested that they were required by an exoglucanase component. Supernatant enzyme preparations contained large amounts of carbohydrate which was separated from most of the cellulase protein by phenyl-Sepharose chromatography. Removal of this carbohydrate, which interfered with protein fractionations, allowed for an activity stain analysis of the supernatant enzyme.

Full text

PDF
304

Images in this article

305Image
on p.305
308Image
on p.308

Selected References

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

  1. Bayer E. A., Setter E., Lamed R. Organization and distribution of the cellulosome in Clostridium thermocellum. J Bacteriol. 1985 Aug;163(2):552–559. doi: 10.1128/jb.163.2.552-559.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Deshpande M. V., Eriksson K. E., Pettersson L. G. An assay for selective determination of exo-1,4,-beta-glucanases in a mixture of cellulolytic enzymes. Anal Biochem. 1984 May 1;138(2):481–487. doi: 10.1016/0003-2697(84)90843-1. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Lamed R., Setter E., Bayer E. A. Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum. J Bacteriol. 1983 Nov;156(2):828–836. doi: 10.1128/jb.156.2.828-836.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. MacKenzie C. R., Bilous D., Johnson K. G. Purification and characterization of an exoglucanase from Streptomyces flavogriseus. Can J Microbiol. 1984 Sep;30(9):1171–1178. doi: 10.1139/m84-183. [DOI] [PubMed] [Google Scholar]
  7. Mackenzie C. R., Bilous D., Patel G. B. Studies on Cellulose Hydrolysis by Acetivibrio cellulolyticus. Appl Environ Microbiol. 1985 Aug;50(2):243–248. doi: 10.1128/aem.50.2.243-248.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ng T. K., Zeikus J. G. Purification and characterization of an endoglucanase (1,4-beta-D-glucan glucanohydrolase) from Clostridium thermocellum. Biochem J. 1981 Nov 1;199(2):341–350. doi: 10.1042/bj1990341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Petre J., Longin R., Millet J. Purification and properties of an endo-beta-1,4-glucanase from Clostridium thermocellum. Biochimie. 1981 Jul;63(7):629–639. doi: 10.1016/s0300-9084(81)80061-2. [DOI] [PubMed] [Google Scholar]
  10. Saddler J. N. Factors limiting the efficiency of cellulase enzymes. Microbiol Sci. 1986 Mar;3(3):84–87. [PubMed] [Google Scholar]
  11. Wood T. M., Wilson C. A., Stewart C. S. Preparation of the cellulase from the cellulolytic anaerobic rumen bacterium Ruminococcus albus and its release from the bacterial cell wall. Biochem J. 1982 Jul 1;205(1):129–137. doi: 10.1042/bj2050129. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES