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Optimization and characterization of cold-active endoglucanase produced by Aspergillus terreus strain AKM-F3 grown on sugarcane bagasse

机译:蔗渣上生长的曲霉曲霉AKM-F3产生的冷活性内切葡聚糖酶的优化与表征

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摘要

The psychrotolerant microfungus Aspergillus terreus strain AKM-F3 (GenBank Accession No. KF417464) was investigated for the production of cold-active endoglucanase using sugarcane bagasse. The effects of different process parameters were studied for hyperproduction. Optimum production was found at 15 degrees C with an initial pH of 5.0 at 120 h. For enzyme production, 1 mM ZnCl2, 0.01% aspartate, 2% lactose, and 3% NaNO3 were regarded as potent inducers. The final production gave an activity 3.764-fold higher than the unoptimized conditions. The molecular weight of purified endoglucanase was 55 kDa, showing a 3.464-fold purification increase with a 23.925% yield having 75.279 U/mg of specific activity. The K m and V max values of the purified enzyme were found to be 0.37 mg/mL and 24.63 U/mg, respectively. The enzyme was more active at 35 degrees C and pH 4.0, whereas more stability was observed at 0 degrees C and pH 7.0. Ca2+, Mn2+, and all tested organic solvents were found to be activators of endoglucanase. EDTA had very low effect, whereas the most active inhibitor was sodium deoxycholate. The present study successfully produced a cold-active endoglucanase with novel properties that can be used for the degradation of cellulosic wastes in cold regions and the economically sound removal of unwanted extra microfibrils from cotton.
机译:研究了使用甘蔗渣生产抗精神病的微真菌曲霉曲霉菌株AKM-F3(GenBank登录号KF417464)来生产冷活性内切葡聚糖酶。研究了不同工艺参数对超生产的影响。发现在15摄氏度,120 h的初始pH为5.0时产量最佳。为了产生酶,将1 mM ZnCl2、0.01%天冬氨酸,2%乳糖和3%NaNO3视为有效的诱导剂。最终生产的活性比未优化的条件高3.764倍。纯化的内切葡聚糖酶的分子量为55 kDa,显示出3.464倍的纯化增加,产率为23.925%,比活性为75.279 U / mg。纯化的酶的K m和V max值分别为0.37 mg / mL和24.63 U / mg。该酶在35摄氏度和pH 4.0下更具活性,而在0摄氏度和pH 7.0下则具有更高的稳定性。发现Ca2 +,Mn2 +和所有测试的有机溶剂都是内切葡聚糖酶的激活剂。 EDTA的作用很低,而活性最高的抑制剂是脱氧胆酸钠。本研究成功生产了一种具有新颖特性的冷活性内切葡聚糖酶,可用于在寒冷地区降解纤维素废料,并从经济上合理地去除棉花中多余的微纤维。

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