链霉菌胞外几丁质酶的制备与筛选

Ibrahim Sani, A. Umar, E. Udeze
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引用次数: 0

摘要

本研究的目的是制备链霉菌胞外几丁质酶,并筛选其对临床分离的白色念珠菌的抗真菌活性。链霉菌从某农田中分离得到;对它们进行了鉴定和筛选,以产生几丁质酶。采用标准方法测定了时间、温度、pH和氮源对几丁质酶产量的影响。采用硫酸铵沉淀法对几丁质酶进行部分纯化。以牛血清白蛋白为标准,分光光度法测定蛋白质浓度。采用琼脂孔扩散法评价几丁质酶对白色念珠菌的抑菌活性。分离到的链霉菌共有3株,均为革兰氏阳性、过氧化氢酶阳性、氧化酶阳性,菌株A和C均为吲哚阳性,只有菌株B为柠檬酸盐阳性。以酵母浸膏为氮源,发酵72 h,温度40℃时几丁质酶产量最高。硫酸铵沉淀(80%)酶活最高,为39.0U/ml。温度为40℃,pH为5.5,底物为1.0%胶体甲壳素时酶活性最高。部分纯化的几丁质酶对白色念珠菌的抑制区为20.11±1.26 mm。与标准药物氟康唑的抑制区(21.42±0.08 mm)比较,差异无统计学意义(P>0.05)。这些发现表明链霉菌在有利条件下产生几丁质酶,该酶可作为抗白色念珠菌和其他含几丁质真菌的抗真菌剂。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Production and Screening of Streptomyces-Extracellular Chitinase
The aim of this research was to produce Streptomyces-extracellular chitinase and screen its antifungal activity on a clinically isolated Candida albicans. The Streptomyces were isolated from an agricultural farmland; they were identified and screened for the chitinase production. Effects of time, temperature, pH and nitrogen sources on the chitinase production were determined using standard methods. Ammonium sulphate precipitation was used to partially purify the chitinase. Protein concentrations were determined spectrophotometrically using bovine serum albumin as standard. Agar-well diffusion method was used to evaluate the antifungal activity of the chitinase on C. albicans. The isolated Streptomyces were of three (3) strains, and all the strains are Gram positive, catalase positive, oxidase positive while, Strain A and C are indole positive and only Strain B is citrate positive. The maximum chitinase production was at 72 h, 40°C and when yeast extract was used as the nitrogen source. Ammonium sulphate (80%) precipitation yielded the highest enzyme activity of 39.0U/ml. The maximum enzyme activity was observed at temperature of 40oC, pH 5.5 and 1.0% colloidal chitin (substrate). The partially purified chitinase showed a zone of inhibition of 20.11 ± 1.26 mm against the Candida albicans. This result has no significant difference (P>0.05) when compared with that of the standard drug (Fluconazole) with 21.42 ± 0.08 mm zone of inhibition. These findings suggest that Streptomyces at favourable conditions produce chitinase, and this enzyme can be used as an antifungal agent on Candida albicans and other chitin containing fungi.
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