UDC 577.112: 577.122: 579.222                                  http://dx.doi.org/10.15407/animbiol18.04.078

KINETIC PARAMETERS OF INULINASE AND LEVANASE ACTIVITY OF RAM RUMEN BACTERIA UNDER THE INFLUENCE OF CLINOPTILOLITE

M. Y. Sabat1, R. Ya. Iskra2

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1Lviv National University of Veterinary Medicine
and Biotechnologies named after S. Z. Gzhytskyj,
50 Pekarska str., Lviv 79010, Ukraine

2Institute of animal biology NAAS,
38 Stus str., Lviv 79034, Ukraine

Fructans are a group of fructose polymers, synthesized by plants and microorganisms. Plant fructans (inulin and flein) have a relatively low degree of polymerization (to 40), and bacterial (levan) — large (up to 1.000.000). Microscopic fungi, yeast and a lot of bacteria, in particular the representatives of the rumen microflora, are capable of producing inulinase, which breaks down a variety of fructane, and levanase, which hydrolyzes the main chain of levan.

We determined the levanase activity of extracellular bacteria extract of rumen in rams which were fed with natural fructan and sorbent of clinoptilolite during twenty one days. Kinetic analysis of the inulinase fructan hydrolysis reaction and levanase was performed in a standard incubation environment modified by the physical-chemical parameters (temperature, pH) and substrates content. We determined the Michaelis constant for the substrate saturation conditions (inulin and levan) and a maximum reaction speed of hydrolysis by the method of Lainuiver-Berk. Temperature (37 °C) and pH optimum (4–6) inulinase and levanase activity was defined.

We have found out that with the increasing concentration of inulin and levan from 0.1 to 0.4–0.5 mM the enzymatic activities of fructanhydrolase monotonously increased to their maximum values and thereafter they were maintained at a constant level. It was found that inulin was metabolized with inulinase at greater speed compared to levan under conditions of saturation with the substrate. Levanase hydrolyzed levan but was inactive concerning inulin. In the presence of clinoptilolite the affinity of both hydrolasesdue to the substrates was increased.

Keywords: FRUCTANS, INULIN, LEVAN, CLINOPTILOLITE, RUMEN MICROBIAL METABOLISM

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