CAJ | 학술논문

目的:利用灰绿曲霉EU7-22发酵产纤维素酶,从中分离到β-葡萄糖苷酶,分析其理化特性,确定其最佳活性条件.方法:灰绿曲霉EU7-22发酵液离心后,上清液经硫酸铵沉淀、Phenyl 6 Fast Flow(high sub)疏水层析和Sephacryl S-200凝胶层析,获得纯化的β-葡萄糖苷酶.结果:纯酶的比活性为5.1 IU／mg,得率为13.89％.SDS-PAGE凝胶电泳分析表明该酶是单亚基蛋白,其分子量为56.2 kDa.在pH 4.0～6.0范围内,β-葡萄糖苷酶具有较高的稳定性,该酶的最适酶促反应pH为5.0.当β-葡萄糖苷酶在温度低于60℃的缓冲液中温育1 h后,酶活损失不大,表现了较好的稳定性;当该酶在温度高于60℃的缓冲液中温育1 h后,酶活迅速丧失.β-葡萄糖苷酶在70℃时具有最大催化活性.结论:灰绿曲霉EU7-22发酵产生的β-葡萄糖苷酶具有较高活性,具有分子量较小、最佳催化温度较高的特点.
목적:이용회록곡매EU7-22발효산섬유소매,종중분리도β-포도당감매,분석기이화특성,학정기최가활성조건.방법:회록곡매EU7-22발효액리심후,상청액경류산안침정、Phenyl 6 Fast Flow(high sub)소수층석화Sephacryl S-200응효층석,획득순화적β-포도당감매.결과:순매적비활성위5.1 IU／mg,득솔위13.89％.SDS-PAGE응효전영분석표명해매시단아기단백,기분자량위56.2 kDa.재pH 4.0～6.0범위내,β-포도당감매구유교고적은정성,해매적최괄매촉반응pH위5.0.당β-포도당감매재온도저우60℃적완충액중온육1 h후,매활손실불대,표현료교호적은정성;당해매재온도고우60℃적완충액중온육1 h후,매활신속상실.β-포도당감매재70℃시구유최대최화활성.결론:회록곡매EU7-22발효산생적β-포도당감매구유교고활성,구유분자량교소、최가최화온도교고적특점.
Objective: β-glucosidase produced by Aspergillus glaucus EU7-22 was purified and characterized. Method: The β-glucosidase of Aspergillus glaucus EU7-22 was purified from ferment supernatant liquid by three steps of purification, ammonium sulfate precipitation (80%, W/V), Phenyl 6 Fast Flow (high sub) column chromatography and Sephacryl S-200 column chromatography, with a specific activity of 5.1 IU/ mg and a yield of 13.89%. Result: The purified β-glucosidase was determined as a monomeric protein with a molecular weight of 56.2 kDa. The enzyme exhibited high stability when it was kept in the buffer at pH 4.0 ～ 6.0 and at the temperature below 60℃.β-glucosidase exhibited its optimal activity at 701. Conclusion: β-glucosidase produced by Aspergillus glaucus EU7-22 with small molecular showed high activity under the optimal conditions.