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高产纤维素酶菌株的筛选
作者:周成 徐磊 肖新 谢越 汪建飞 马忠友 来源:《安徽农学通报》2016年第08期
摘 要:该研究从农田土壤中分离得到67株细菌,并从中选出一株高产纤维素酶菌株。将这些菌株接种到纤维素培养基上,在pH 5.5和28℃的条件下,利用刚果红染色溶液进行染色后,测其水解透明圈与菌落的比值的大小,进行初步筛选。将这些初筛的菌株接种到培养基中进行摇床培养后,制得粗酶液,并分析羧甲基纤维素酶(CMC)活力测定和滤纸酶(FP)活力及β-葡萄糖苷酶(BG)。在不同温度的条件下,对纤维素酶活力测定,最终筛选出曲霉A25(Aspergillus sp.)这一株菌株,最适酶活温度为50℃。产纤维素酶酶活力分别为:CMC酶活达2 340.92 U/mL;FP酶活达2.66U/mL;BG酶活达164.72U/mL。 关键词:羧甲基纤维素酶;滤纸酶;β-葡萄糖苷酶;曲霉A25 中图分类号 S154 文献标识码 A 文章编号 1007-7731(2016)08-22-04
Abstract:67 strains of soil bacteria were isolated form farmland,from which a high-yielding cellulase strain was screened.These strains were inoculated on the cellulose medium,and a preliminary screening was conducted,in which we measured hydrolysis transparent circle and the ratio of the size of the colony under pH5.5 and 28℃ conditions by using the Congo Red
staining.Furthermore,these selected strains were cultured,and a crude enzyme liquid was extracted for analyzing the activities of carboxymethyl cellulose(CMC),filter paper enzyme(FP)and β-glucosidase(BG).Under different temperature conditions,the activity of cellulase was
determined,and ultimately this one strain of Aspergillus A25 was obtained.Its great activity is under the condition of optimum temperature for 50 ℃.Producing cellulose enzyme activity was as following:CMC amounted to 2 340.92 U/mL,FP activity amounted to 2.66 U/mL,and BG activity amounted to 164.72 U/mL.
Key words:Cerboxymethl cellulose;Filter paper enzyme;β- glucosidase;Aspergillus A25 纤维素物质是地球上含量最丰富的碳水化合物[1],而目前人类对纤维素的开发与利用还非常有限,因此如何更有效地开发和利用纤维素资源已成为当今世界的热门课题之一。目前,对纤维素的降解利用主要是用酸碱处理等化学手段,以及汽爆及蒸汽加热等物理手段[2]。生物法由于对设备要求低,分解后的产物易回收利用,以及对环境污染较小等特点而备受重视。纤维素是一个复杂酶系,根据其功能的不同可分为3类:作用于纤维素非结晶区的内切葡聚糖酶(CMC);作用于无定型区切割糖苷键的外切葡聚糖酶(FP);以及作用于纤维二糖的β-葡萄糖苷酶(BG)[3-5]。协同作用才能将结构复杂的天然纤维降解。纤维素酶在食品、洗涤、纺织、饲料、造纸等方面具有广泛的应用和发展前景[5]。通过对本实验室保存的67株菌株进行初筛和复筛,A25这一株菌株是本次实验所筛选的,具有较高降解纤维素活力的菌株,
高产纤维素酶菌株的筛选



