椰果内生枯草芽孢杆菌YZ-21产β-甘露聚糖酶的纯化及酶学性质研究  

Purification and properties of β-mannanse produced by Bacillus subtilis YZ-21 in coconut fruit

作　　者：张建新 郭祥瑞 穆广亚 冯军厂 常绪路 Zhang Jianxin;Guo Xiangrui;Mu Guangya;Feng junchang;Chang Xulu(College of Fisheries,Henan Normal University,Xinxiang 453007,China)

机构地区：[1]河南师范大学水产学院,河南新乡453007

出　　处：《河南师范大学学报（自然科学版）》2020年第3期95-102,共8页Journal of Henan Normal University(Natural Science Edition)

基　　金：国家自然科学青年基金(31902361);河南省重点科技攻关(182102110011);河南省自然科学基金(182300410098).

摘　　要：[目的]分离纯化来源于内生枯草芽孢杆菌YZ-21的β-甘露聚糖酶,并研究其酶学性质.[方法]采用乙醇沉淀法、硫酸铵盐析法、聚乙二醇(PEG)/硫酸铵[(NH 4)2SO 4]双水相体系进行分离与纯化,在此基础研究它的酶学性质.[结果]比较3种纯化方法,最优方法为双水相萃取法,在双水相体系为16%(NH 4)2SO 4、18%PEG2000、2%NaCl条件下,纯化倍数最高可以达到3.06,酶回收率达到99.26%;酶学性质研究表明,β-甘露聚糖酶的最适反应pH为7.0,最适的反应温度是40℃,pH稳定性在3.0~7.0,热稳定范围在35~55℃之间;Ca^2+,Ba^2+,Mg^2+,K^+,Co^2+对酶都有激活作用,而Ca^2+激活作用最强,Cu^2+,Na^+,Zn^2+,Mn^2+,EDTA对酶有抑制作用.[结论]利用双水相萃取法较好纯化了β-甘露聚糖酶,酶学性质良好,可以广泛应用于饲料添加剂、纸浆漂白和食品加工等领域.[Objective]The aim of this study is to isolate and purify β-mannanase from endogenous Bacillus subtilis YZ-21,and further study its enzymatic properties.[Method]Ethanol precipitation method,ammonium sulfate salting out method,polyethylene glycol(PEG)/ammonium sulfate[(NH 4)2SO 4]dual aqueous phase system were used for separation and purification.On this basis,its enzymatic properties were studied.[Result]The optimal purification method was finally obtained by the two-aqueous phase extraction method.The optimal two-aqueous phase system was 16%(NH 4)2SO 4,18%PEG 2000,and 2% NaCl.Under this condition,the purification factor could reach 3.06,the enzyme recovery rate reached 99.26%.The optimal reaction pH of β-mannanase is 7.0,the optimal reaction temperature is 40℃,the pH stability range is 3.0-7.0,and the thermal stability range is below 60℃;Ca^2+,Ba^2+,Mg^2+,K^+,Co^2+ have activation effects,while Ca^2+ has the strongest activation effect,and Cu^2+,Na^+,Zn^2+,Mn^2+ and EDTA have inhibitory effects on enzymes.[Conclusion]Using aqueous two-phase extraction,β-mannanase was preferably purified,which has good enzymatic properties,can be widely used in feed additives,pulp bleaching,food processing and the like.

关 键 词：Β-甘露聚糖酶 枯草芽孢杆菌 分离纯化 酶学性质 

分 类 号：TQ464[化学工程—制药化工]