Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Carboxymethylcellulase production by the anaerobic rumen fungusNeocallimastix sp. GMLF7

  • 71 Accesses

  • 9 Citations

Abstract

Anaerobic fungi have highly active fibrolytic enzymes and these enzymes are attractive for scientific research. We isolated a ruminal fungus of the genusNeocallimastix sp., named GMLF7, which could survive on a variety of cellulosic material such as carboxymethylcellulose (CMC), fibrous cellulose, avicel and wheat straw. Carboxymethylcellulase production was investigated with the above carbon sources and high CMCase activity was obtained with CMC (73.75 U/ml), fibrous cellulose (72.68 U/ml) and avicel (70.03 U/ml). While growth temperature of the microorganism was 39°C, for CMCase activity optimal temperature was 50°C and optimal pH 6.5. Enzyme reached maximum activity in 60–180 min at 50°C. Substrate concentration also affected the enzyme activity which was increased more than 2 fold with 40 mg/ml CMC. Activity was measured in presence of various divalent ions and reagents, and Co2+ and DTT has a positive effect on the enzyme activity.

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

References

  1. Akin D.E., Gordon G.L.R., Hogan J.P. (1983). Rumen bacterial and fungal degradation ofDigitaria pentzii grown with or without sulfur. Appl. Environ. Microbiol., 46: 738–748.

  2. Akin D.E. (1994). Ultrastructure of plant cell-walls degraded by anaerobic fungi. In: Mountfort D.O., Orpin C.G., Eds, Anaerobic Fungi, Marcel Dekker, Inc., New York, pp. 169–190.

  3. Barichievich E.M., Calza R.E. (1990). Supernatant protein and cellulase activities of the anaerobic ruminal fungusNeocallimastix frontalis EB188. Appl. Environ. Microbiol., 56: 43–48.

  4. Bhat M.K. (2000). Cellulases and related enzymes in biotechnology. Biotech. Adv., 18: 355–383.

  5. Borneman W.S., Akin D.E., Ljungdahl L.G. (1989). Fermentation products and plant cell wall-degrading enzymes produced by monocentric and polycentric anaerobic ruminal fungi. Appl. Environ. Microbiol., 55: 1067–1073.

  6. Garg S.K., Neelakantan S. (1981). Effect of cultural factors on cellulase activity and protein production byAspergillus terreus. Biotechnol Bioeng., 23: 1653–1659.

  7. Harhangi H.R., Akhmanovab A., Steenbakkers P.J.M., Jetten M.S.M., Drift C., Camp H.J.B. (2003). Genomic DNA analysis of genes encoding (hemi-)cellulolytic enzymes of the anaerobic fungusPiromyces sp. E2. Gene, 314: 73–80.

  8. Hungate R.E. (1969). A roll tube method for the cultivation of strict anaerobes. Methods In Microbiology, Academic Press, London.

  9. Li X.L., Chen, H., Ljungdahl L.G. (1997). Two cellulases, CelA and CelC, from the polycentric anaerobic fungusOrpinomyces strain PC-2 contain N-terminal docking domains for a cellulase-hemicellulase complex. Appl. Environ. Microbiol., 63: 4721–4728.

  10. Lin L.L., Chyau C.C., Hsu W.H. (1998). Production and properties of a raw-starch-degrading amylase from the thermophilic and alkaliphilicBacillus sp. TS-23. Biotechnol. Appl. Biochem., 28: 61–68.

  11. Liu J.R., Yu B., Liu F.H., Cheng K.J., Zhao X. (2005). Expression of rumen microbial fibrolytic enzyme genes in probioticLactobacillus reuteri. Appl. Environ. Microbiol., 71: 6769–6775.

  12. Lowe S.E., Theodorou M.K., Trinci A.P.J. (1987) Cellulases and xylanases of an anaerobic rumen fungus grown on wheat straw, wheat straw holocellulose, cellulose, and xylan. Appl. Environ. Microbiol., 53: 1216–1223.

  13. Lynd L.R., Weimer P.J., Zyl W.H., Pretorius I.S. (2002). Microbial cellulose utilization: fundamentals and biotechnology. Microbiol. Mol. Biol. Rev., 66: 506–77.

  14. Mawadza C., Hatti-Kaul R., Zvauya R., Mattiasson B. (2000). Purification and characterization of cellulases produced by twoBacillus strains. J. Biotech., 83: 177–187.

  15. Miller G.L. (1959). Use of dinitrosalicylic reagent for the determination of reducing sugars. Anal. Chem., 31: 426–428.

  16. Mountfort D.O., Asher R.A. (1985). Production and regulation of cellulase by two strains of the rumen anaerobic fungusNeocalliniastix frontalis. Appl. Environ. Microbiol., 49: 1314–1322.

  17. Mountfort D.O., Asher R.A. (1989). Production of xylanase by the ruminal anaerobic fungusNeocallimasix frontalis. Appl. Environ. Microbiol., 55: 1016–1022.

  18. Niehaus F., Bertoldo C., Kahler M., Antranikian G. (1999). Extremophiles as a source of novel enzymes for industrial application. Appl. Microbiol. Biotech. 51: 711–729.

  19. Niranjane A.P., Madhou P., Stevenson T.W. (2007). The effect of carbohydrate carbon sources on the production of cellulase byPhlebia gigantean. Enzyme Microb. Tech., 40: 1464–1468.

  20. Oksanen T., Pere J., Paavilainen L., Buchert J., Viikari L. (2000). Treatment of recycled kraft pulps withTrichoderma reesei hemicellulases and cellulases. J. Biotech., 78: 39–48.

  21. Orpin C.G. (1975). Studies on the rumen flagellateNeocallimastix frontalis. J. Gen. Microbiol., 91: 249–262.

  22. Orpin C.G., Letcher A.J. (1979). Utilization of cellulose, starch, xylan, and other hemicelluloses for growth by the rumen phycomycete,Neocallimastix frontalis. Curr. Microbiol., 3: 121–124.

  23. Qiu X., Selinger B., Yanke L.J., Cheng K.J. (2000). Isolation and analysis of two cellulase cDNAs fromOrpinomyces joyonii. Gene, 245: 119–126.

  24. Shin C.S., Lee J.P., Lee J.S., Park S.C. (2000). Enzyme production ofTrichoderma reseei Rut C-30 on various lignocellulosic substrates. Appl. Biochem. Biotechnol., 84–86: 237–245.

  25. Steenbakkers P.J.M., Harhangi H.R., Bosscher M.W., Van Der Hooft M.M.C., Keltjens J.T., Van Der Drift C., Vogels G.D., Op Den Camp H.J.M. (2003). β-Glucosidase in cellulosome of the anaerobic fungusPiromyces sp. strain E2 is a family 3 glycoside hydrolase. Biochem. J., 370: 963–970.

  26. Trinci A.P.J., Davies D.R., Gull K., Lawrence M.I., Nielsen B.B., Rickers A., Theodorou M.A. (1994). Anaerobic fungi in herbivorous animals. Mycol. Res., 98: 129–152.

  27. Wallace R.J. (1994). Ruminal microbiology, biotechnology and ruminant nutrition: progress and problems. J. Anim. Sci., 72: 2992–3003.

Download references

Author information

Correspondence to Ugur Comlekcioglu.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Comlekcioglu, U., Akyol, I., Ozkose, E. et al. Carboxymethylcellulase production by the anaerobic rumen fungusNeocallimastix sp. GMLF7. Ann. Microbiol. 58, 115–119 (2008). https://doi.org/10.1007/BF03179454

Download citation

Key words

  • anaerobic fungi
  • Neocallimastix sp.
  • CMCase
  • metal ions