Abstract
P. ruminis strain 3 was isolated from the ovine rumen and identified on the basis of comparison of its 16S rRNA gene with GenBank. The bacterium was able to grow on Timothy grass fructan, inulin, sucrose, fructose and glucose as a sole carbon source, reaching absorbance of population in a range of 0.4–1.2. During 1 d the bacteria exhausted 92–97 % of initial dose of saccharides except for inulin (its utilization did not exceed 33 %). The bacterial cell extract catalyzed the degradation of Timothy grass fructan, inulin and sucrose in relation to carbon source present in growth medium. Molecular filtration on Sephadex G-150, polyacrylamide gel electrophoresis combined with zymography technique and TLC was used to identify enzymes responsible for the digestion of sucrose and both polymers of fructose. Two specific endolevanases (EC 3.2.1.65), nonspecific β-fructofuranosidase (EC 3.2.1.80 and/or EC 3.2.1.26) and sucrose phosphorylase (EC 2.4.1.7) were detected in cell-free extract from bacteria grown on Timothy grass fructan.
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Gabriel O., Wang S.-F.: Determination of enzymatic activity in polyacrylamide gels. Analyt.Biochem.27, 545–554 (1969).
Guczyńska W.: Substrate preferences of some of rumen bacteria in metabolism of sugars. PhD Thesis (in Polish). The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences. Jabłonna near Warsaw, Poland (1999).
Kasperowicz A., Michałowski T.: Assessment of fructanolytic activities in the rumen bacterium Treponema saccharophilum strain S. J.Appl.Microbiol.92, 140–146 (2002).
Klaus R., Fisher W.: Quantitative thin-layer chromatography of sugars, sugar acids, and polyalcohols, pp. 159–180 in W.A. Wood, S.T. Kellogg (Eds): Methods in Enzymology, Vol. 160. Academic Press-Harcourt Brace Jovanovich Publishers, San Diego 1988.
Lüscher M., Frehner M., Nösberger J.I.: Purification and some properties of fructan: fructan-fructosyl transferase from dandelion (Taraxacum officinale WEBER). New Phytol.123, 437–442 (1993).
Miller G.L., Blum R., Glennon W.E., Burton A.L.: Measurement of carboxymethylcellulase activity. Analyt.Chem.2, 127–132 (1960).
Purich D.L., Allison R.D.: The Enzyme Reference. A Comprehensive Guide to Enzyme Nomenclature, Reactions and Methods. Academic Press, San Diego 2002.
Ziołecki A., Guczyńska W., Wojciechowicz M.: Some rumen bacteria degrading fructan. Lett.Appl.Microbiol.15, 244–247 (1992).
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Kasperowicz, A., Stan-Glasek, K., Guczynska, W. et al. Fructanolytic and saccharolytic enzymes of the rumen bacterium Pseudobutyrivibrio ruminis strain 3 — preliminary study . Folia Microbiol 55, 329–331 (2010). https://doi.org/10.1007/s12223-010-0051-4
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DOI: https://doi.org/10.1007/s12223-010-0051-4