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首页> 外文期刊>African Journal of Biotechnology >Purification and characterization of xylanase from Aspergillus fumigatus isolated from soil
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Purification and characterization of xylanase from Aspergillus fumigatus isolated from soil

机译：从土壤中分离出的烟曲霉木聚糖酶的纯化和鉴定

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The objectives of the present study were to purify and characterize xylanase enzyme from the fungus obtained from soil. A total of 40 fungi were isolated from 25 soil samples collected after primary screening on Potato Dextrose Agar. In the secondary screening (malt extract agar, 0.5% birch wood xylan), based on the diameter of the clear zone, the fungus was identified as?Aspergillus fumigatus?by microbial type culture collection (MTCC), Chandigarh, India and was selected for xylanase enzyme production in solid state fermentation using wheat bran. Xylanase was subjected to a three-step purification scheme involving ammonium sulphate precipitation, gel filtration chromatography and anion exchange chromatography. Purity was verified by running the extracted protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a single band was observed. When compared with the standard wide range protein molecular markers on SDS-PAGE, it was found to have a molecular weight of 43 KDa. The Km?and Vmax?value of xylanase was 3.12 mg/ml and 2857 μmol/min/mg protein as obtained from a Lineweaver-Burk plot. The optimal temperature and pH was found to be 30 and 10°C, respectively. After 4 h of incubation, enzyme retained 100% activity at 30°C. Xylanase was incubated at various pH levels (2 to 12) for 4 h at 30°C, and the residual activity was measured. More than 65% of the original activity was retained at pH ranging from 4 to 10 after 4 h.
机译：本研究的目的是从土壤获得的真菌中纯化和表征木聚糖酶。在马铃薯右旋糖琼脂上初步筛选后，从收集的25个土壤样品中分离出总共40种真菌。在二次筛选（麦芽提取物琼脂，0.5％桦木木聚糖）中，根据透明区的直径，该真菌被印度昌迪加尔的微生物类型培养物收集中心（MTCC）鉴定为“烟曲霉”，并被选作使用麦麸固态发酵生产木聚糖酶。对木聚糖酶进行三步纯化方案，包括硫酸铵沉淀，凝胶过滤色谱和阴离子交换色谱。通过在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）上运行提取的蛋白来验证纯度，并观察到一条条带。当与SDS-PAGE上的标准宽范围蛋白质分子标记进行比较时，发现其分子量为43 KDa。从Lineweaver-Burk图获得的木聚糖酶的Km？和Vmax？值为3.12 mg / ml和2857μmol/ min / mg蛋白。发现最佳温度和pH分别为30和10℃。温育4小时后，酶在30℃下保持100％活性。木聚糖酶在各种pH值（2至12）下于30°C孵育4小时，并测量残留活性。 4小时后，pH在4至10范围内保留了超过65％的原始活性。

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