Current Microbiology

, Volume 23, Issue 3, pp 175–180 | Cite as

Nascent synthesis and secretion of cellobiase inNeocallimastix frontalis EB188

  • R. E. Calza


De novo synthesis and the secretion of cellobiase fromNeocallimastix frontalis EB188 were studied. Cellobiase was secreted rapidly in cellulose switch cultures. Chemical inhibition of protein synthesis and radiotracer studies suggested secretion was dependent on nascent protein synthesis. Inhibitors of protein glycosylation also inhibited secretion. An 85,000 dalton protein (and several others) represented the principal differences in de novo synthesis between cellulose- and glucose-switched cultures. Concentrations of the 85,000 daltons protein increased with culture incubation time and ultimately accounted for 7% of the total protein secreted. This protein was purified by gel electrophoresis separations and was determined to be a cellobiase. Secretion of this cellobiase was correlated with its radiolabeling. The possibility of cellobiase induction representing a specialized gene control system inNeocallimastix frontalis EB188 is discussed.


Cellulose Control System Electrophoresis Protein Synthesis Incubation Time 
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Literature Cited

  1. 1.
    Barichievich EB, Calza RE (1990) Supernatant protein and cellulase activities of the anaerobic ruminal fungusNeocallimastix frontalis EB188. Appl Environ Microbiol 56:43–48PubMedGoogle Scholar
  2. 2.
    Barichievich EB, Calza RE (1990) Media carbon induction of extracellular cellulase activities inNeocallimastix frontalis EB188. Curr Microbiol 20:265–271Google Scholar
  3. 3.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72:248–252PubMedGoogle Scholar
  4. 4.
    Calza RE (1990) Regulation of protein and cellulase excretion in the ruminal fungusNeocallimastix frontalis EB188. 21:109–115Google Scholar
  5. 5.
    Calza RE, Schroeder AL (1983) Release of high molecular weight DNA fromNeurospora crassa using enzymic digestions. J Gen Microbiol 129:413–422PubMedGoogle Scholar
  6. 6.
    Calza RE, Irwin DC, Wilson DB (1985) Purification and characterization of two B-1,4 endoglucanases fromThermomonospora fusca. Biochemistry 24:7797–7804Google Scholar
  7. 7.
    Hungate RE (1969) A roll-tube method for the cultivation of strict anaerobes. In: Norris JR, Ribbons DW (eds). Methods Microbiol 3B:117–132Google Scholar
  8. 8.
    Joblin KN (1981) Isolation, enumeration, and maintenance of rumen anaerobic fungi in roll tubes. Appl Environ Microbiol 42:1119–1122Google Scholar
  9. 9.
    Jue CL, Lipke PN (1985) Determination of reducing sugars in the nanomole range with tetrazolium blue. J Biochem Biophys Methods 11:109–116PubMedGoogle Scholar
  10. 10.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedGoogle Scholar
  11. 11.
    Lowe SE, Theodorou ML, Trinci APJ, Hespell RB (1985) Growth of anaerobic rumen fungi in defined and semi-defined media lacking rumen fluid. J Gen Microbiol 131:2225–2229Google Scholar
  12. 12.
    Maurer HR (1971) In: (eds) Discontinuous electrophoresis and related techniques of PAGE. New York: Walter de Gruyter & Co., pp 4–52Google Scholar
  13. 13.
    Mountfort DO, Asher RA (1983) Role of catabolite regulatory mechanisms in control of carbohydrate utilization by the rumen anaerobic fungusNeocallimastix frontalis. Appl Environ Microbiol 46:1331–1338PubMedGoogle Scholar
  14. 14.
    Mountfort DO, Asher RA (1985) Production and regulation of cellulase by two strains of the rumen anaerobic fungusNeocallimastix frontalis. Appl Environ Microbiol 49:1314–1322PubMedGoogle Scholar
  15. 15.
    Orpin CG (1977) The rumen flagellatePiromonas communis: its life-history and invasion of plant material in the rumen. J Gen Microbiol 99:107–117PubMedGoogle Scholar
  16. 16.
    Sadana JC, Patil RV, Shewale JG (1988) Beta-d-glucosidase fromSelerotium rolfsii. Methods Enzymol 160 (A):424–431Google Scholar
  17. 17.
    Suzuki H, Terada T (1988) Removal of dodecyl sulfate from protein solution. Anal Biochem 172:259–263PubMedGoogle Scholar
  18. 18.
    Whistler RL, Wolfram ML (1962) Determination of reducing sugars and carbohydrates. Methods Carbohydrate Chem 1:389–390Google Scholar
  19. 19.
    Williams AG, Orpin CG (1987) Polysaccharide-degrading enzymes formed by the three species of anaerobic rumen fungi grown on a range of carbohydrate substrates. Can J Microbiol 33:419–426Google Scholar
  20. 20.
    Wood TM, Wilson CA, McCrae SI, Joblin KN (1986) A highly active extracellular cellulase from the anaerobic rumen fungusNeocallimastix frontalis. FEMS Microbiol Lett 34:37–40Google Scholar
  21. 21.
    Wood TM, McCrae SI, Wilson CA, Bhat KM, Gow LA (1988) In: Aubert JP, Bequin P, Millet J (eds) Biochemistry and genetics of cellulose degradation. London: Academic Press, Inc. (London) Ltd., pp 31–52Google Scholar
  22. 22.
    Wray W, Baulikas T, Wray VP, Hancock R (1981) Silver staining of proteins in polyacrylamide gels. Anal Biochem 118:197–203PubMedGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 1991

Authors and Affiliations

  • R. E. Calza
    • 1
  1. 1.Department of Animal SciencesWashington State UniversityPullmanUSA

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