PL EN


Preferences help
enabled [disable] Abstract
Number of results
2010 | 15 | 189 - 196
Article title

ISOLATION AND PURIFICATION OF CHITINOLYTIC ENZYMES OF RUMEN CILIATES Eudiplodinium maggii

Content
Title variants
Languages of publication
EN
Abstracts
EN
Results of the earlier studies suggested an involvement of ciliates Eudiplodinium maggii in the digestion and metabolism of chitin in the rumen. In the presented paper we described the results on the preliminary identification and characterization of chitinolytic enzymes of this ciliate as well as the method of their purification. The protozoal crude enzyme preparation was used as source of enzymes, whereas the molecular filtration on Sephadex G-150 (single step method) or separation of protein by precipitation with ammonium sulfate followed by molecular filtration (two step method) were applied to purify the identified enzymes. The identification studies resulted in the detection of endochitinase, exochitinase and N-acetyl-D-glucosaminidase. The highest activity of identified enzymes were obtained in 4.0 - 4.5 pH and at 45 - 50 °C. Results of the comparative study on purification procedures showed that the single-step method enabled us to obtain enzymes of higher purity and higher activity than the two-step purification method.
Publisher

Year
Volume
15
Pages
189 - 196
Physical description
Contributors
author
  • The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences, r.miltko@ifzz.pan.pl
  • The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences
  • The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences
  • The Kielanowski Institute of Animal Physiology and Nutrition Polish Academy of Sciences
References
  • Bełżecki G., Miltko R., Michałowski T., Šimůnek J., Kopečný J.; (2008) Chitinolytic activity of the rumen ciliates Diploplastron affine. Folia Microbiol. 53, pp. 201-203.
  • Coleman G. S., Davies J. I., Cash M. A.; (1972) The cultivation of rumen ciliates Epidinium ecaudatum and Polyplastron multivesiculatum in vitro. J. Gen. Microbiol. 75, pp. 509-521.
  • Dogiel V. A.; (1927) Monographie der Familie Ophryoscolecidae. Arch. Protistenk. 59, pp. 1-288.
  • Hungate R. E.; (1942) The culture of Eudiplpdinium neglectum with experiments on the digestion of cellulose. Biol. Bull. 83, pp. 303-319.
  • Kopecný J., Hodrová B.; (2000) Chitinolytic enzymes produced by ovine rumen bacteria. Folia Microbiol., 45, pp. 465-468.
  • Michałowski T., Harmeyer J., Bełżecki G.; (1999) The importance of washing the omasum for successful defaunation of sheep. J. Anim. Feed. Sci. 8, pp. 611-619.
  • Michałowski T.; (1979) A simple system for continuous culture of rumen ciliates. Bull. Pol. Acad. Sci. Biol. 27, pp. 581-583.
  • Miller G. L., Blum R., Glennon W. E., Butron A. L.; (1960) Measurement of carboxymethylcellulase activity. Anal. Biochem. 2, pp. 127-132.
  • Morgavi D. P., Sakurada M., Tomita Y., Onodera R.; (1994) Presence in rumen bacteria and protozoal populations of enzymes capable of degrading fungal cell walls. Microbiology 140, pp. 631-636.
  • Novotná Z, Fliegerová K, Simůnek J.; (2008) Characterization of chitinases of polycentric anaerobic rumen fungi. Folia Microbiol. 53, pp. 241-245.
  • Orpin C. G.; (1975) Studies on the rumen flagellate Neocalimastix frontalis. J. Gen. Microbiol. 84, pp. 395-398.
  • Reguera G. Leschine S. B.; (2003) Biochemical and genetic characterization of ChiA, the major enzyme component for the solubilization of chitin by Cellulomonas uda. Arch Microbiol. 180, pp. 434-443.
  • Sakurada M.; Morgavi D. P.; Ushirone N., Komatani K., Tomita Y., Onodera R.; (1998) Purification and characteristics of membrane-bound chitinase of anaerobic ruminal fungus Piromyces communis OTS1. Current Microbiology. 37, pp. 60-63.
  • Takeshi T., Shinsuke F., Shingo N., Toshiaki F., Masahiro T., Tadayuki I.; (1999) A Unique Chitinase with Dual Active Sites and Triple Substrate Binding Sites from the Hyperthermophilic Archaeon Pyrococcus kodakaraensis KOD1. Appl. Environ Microbiol. 63, pp. 5338-5344.
  • Wirth S. J., Walh G. A.; (1990) Day-label substrates for the assay and detection of chitinase and lysozyme activity. J. Microbial Method. 12, pp. 197-205.
  • Yem D. W., Wu H. C.; (1976) Purification and properties of β-N-Acetyloglucosoamidase from Escherichia coli. J. Bacteriol. 125, pp. 324-331
Document Type
article
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.psjd-d5f5cf6e-937b-434d-9e69-a17faef098af
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.