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瘤胃纤毛虫 Eudiplodinium maggii 的几丁质酶。

Chitinolytic enzymes of the rumen ciliate Eudiplodinium maggii.

机构信息

The Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Instytucka 3, 05-110, Jabłonna near Warsaw, Poland.

出版信息

Folia Microbiol (Praha). 2012 Jul;57(4):317-9. doi: 10.1007/s12223-012-0133-6. Epub 2012 Apr 13.

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.

摘要

纤毛虫能够消化几丁质这一能力已得到明确的认可。我们研究了反刍动物纤毛虫真毛滴虫的几丁质分解系统。这些纤毛虫在选择性饲养的绵羊中生长。它们从瘤胃中分离出来,并通过沉淀进行纯化。在使用抗生素孵育纤毛虫之后,制备了粗酶制剂,以减少其与细胞内细菌的污染。通过定量测定特定底物释放的产物,检查了特定酶的活性。据指出,检测到的活性的最佳条件在 pH4.5 到 5.5 之间,在 45 到 55°C 之间变化。β-N-乙酰氨基葡萄糖苷酶被发现是活性最高的酶(每毫克蛋白每小时释放 4.2 μmol 的产物)。内切几丁质酶和外切几丁质酶的活性几乎是β-N-乙酰氨基葡萄糖苷酶的两倍。同工酶研究表明,在检查的制剂中存在两种内切几丁质酶、两种外切几丁质酶和两种β-N-乙酰氨基葡萄糖苷酶。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a0f4/3389239/f2f69b82e344/12223_2012_133_Fig1_HTML.jpg

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