裂叶菌Bcc26414纤维素二糖脱氢酶的纯化及特性研究

Vinita Yadav, Sadhana Nighojkar
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引用次数: 0

摘要

CDH是一种由木材分解真菌产生的酶,在生物传感器生产、生物修复和生物医学工业中有着广泛的应用。本研究采用硫酸铵沉淀法、deae -纤维素层析法和Sephadex G-200层析法对Schizophyllum commune BCC26414中的CDH进行纯化。纯化倍数为65.81,比活性为1612.34 U/mg。CDH的纯度和分子量通过native和SDS PAGE进行了验证。最佳温度为30℃,pH为5℃。纯化后的CDH在较宽的pH范围(3.5 ~ 6.5)内表现出24小时的稳定性,在40°C下保持完全的活性,在50°C下观察150分钟后活性降低。KCl、MgSO4、ZnSO4和NiCl2在5 mM浓度下增强CDH活性,HgCl2和CuSO4抑制酶活性。CDH对乳糖的动力学常数、Km和Vmax分别为125 mM和13.26 U/ml。纯化后的CDH可用于各种商业用途。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cellobiose Dehydrogenase from Schizophyllum commune Bcc26414: Purification and Characterization
CDH, an enzyme produced by wood-decomposing fungi, has diverse applications in biosensor production, bioremediation, and biomedical industries. In this study, CDH from Schizophyllum commune BCC26414 was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and Sephadex G-200 chromatography. The purification fold achieved was 65.81 with a specific activity of 1612.34 U/mg. The purity and molecular weight of CDH was confirmed using native and SDS PAGE. Optimal temperature and pH were found to be 30°C and 5, respectively. The purified CDH exhibited stability over a wide pH range (3.5 to 6.5) for 24 hrs and retained complete activity at 40°C, with reduced activity at 50°C when observed for 150 min. KCl, MgSO4, ZnSO4, and NiCl2 at a concentration of 5 mM enhanced CDH activity and HgCl2 and CuSO4 inhibited the enzyme activity. The kinetic constants, Km and Vmax of CDH for lactose were observed to be 125 mM and 13.26 U/ml, respectively. The purified CDH may be utilized commercially in various applications.
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