Abstract
The ability of rumen ciliates to digest chitin is clearly recognized. We investigated the chitinolytic system of the rumen ciliate Eudiplodinium maggii. The ciliates were grown in a selectively faunated sheep. They were isolated from the rumen and purified by sedimentation. A crude enzyme preparation was prepared following incubation of ciliates with antibiotics. This was done in order to reduce their contamination with intracellular bacteria. The activity of particular enzymes was examined by quantification of the products released from specific substrates. It was stated that the optimum conditions for the detected activities varied between 4.5 and 5.5 pH, and 45 and 55 °C. β-N-Acetylglucosaminidase was found as an enzyme of the highest activity (4.2 μmol/l released product per mg protein per h). The activities of endochitinase and exochitinase were almost two times lower than that of β-N-acetylglucosaminidase. Zymographic studies revealed the presence of two endochitinases, two exochitinases and two β-N-acetylglucosaminidases in the examined preparation.
Highlights
Rumen ophryoscolecid protozoa engulf readily the fungal zoospores which are rich in chitin (Williams and Coleman 1997; Lee et al 2001)
We already found that the ciliates Eudiplodinium maggii digest and ferment chitin (Miltko et al 2010)
We showed that E. maggii ciliates possess endochitinase, exochitinase and β-N-acetylglucosaminidase which were responsible for the breakdown of chitin
Summary
Rumen ophryoscolecid protozoa engulf readily the fungal zoospores which are rich in chitin (Williams and Coleman 1997; Lee et al 2001). We already found that the ciliates Eudiplodinium maggii digest and ferment chitin (Miltko et al 2010). No information is available on chitinolytic enzymes of this species of rumen protozoa. The objective of this study was to identify and characterize chitinolytic enzymes of this species of ciliates
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