内切型菊粉酶酶学性质的研究  

Fermentation production technique optimalization of endo-inulinase by response surface analysis

作　　者：王春[1] 曹泽虹[1] 董玉玮[1] 高兆建[1] 陈阔[1] 李雪晴[1] 王娟[1] 

机构地区：[1]徐州工程学院食品(生物)工程学院,江苏省食品资源开发与质量安全重点建设实验室,江苏省食品与生物工程实验中心,徐州221008

出　　处：《中国食品添加剂》2015年第3期119-126,共8页China Food Additives

基　　金：江苏省高校自然科学研究计划项目(07KJD550202);徐州市科技发展指导性计划项目(XZZD1204);徐州工程学院理工培育项目(XKY2011109);国家级大学生实践创新训练计划项目(XCX2014039)

摘　　要：以食品与生物工程实验中心酶工程实验室保藏的黑曲霉菌种作为菌源,利用黑曲霉细胞深层发酵法生产内切型菊粉酶。通过测定发酵液酶活,从20株黑曲霉菌株中筛选得到产菊粉酶活力最高的编号为E133131的一株黑曲霉菌株,酶活为0.66U/mL。对黑曲霉E133131号菌株所产内切型菊粉酶进行了酶学性质的研究,得到以下结论：最适pH为4.5;最适温度为60℃;最佳底物浓度为4mmol/L;米氏常数（Km）为1.434mmol/L;Ca^2＋对内切型菊粉酶有激活作用,当加入浓度为1.5mmol/L的Ca^2＋时,使酶活从0.66U/mL提高到1.09U/mL;Mn2＋、Cu^2＋和Mg2＋对内切型菊粉酶有抑制作用,其中Cu^2＋的抑制作用最大,使酶活从0.66U/mL降低到0.095U/mL;Na^＋、K＋、Zn2＋、Fe2＋、Mg2＋对内切型菊粉酶的酶活影响不大;pH在4.5-8.0范围内,温度在50-60℃之间,内切型菊粉酶的酶活力较稳定。This experiment regards aspergillus strains preserved by enzyme engineering laboratory as a source of bacteria,the aspergillus niger type cutting inside inulinase produced by enzymatic properties were studied. Of 20 strains of aspergillus strains screening in the first place,the enzyme activity can be determined through fermentation cultivation,finally selected one of the highest enzyme activity strain E133131,the enzyme activity of 0.66U/m L. Researching aspergillus niger E133131 strains produced inulinase enzymology properties,get the following conclusion： the optimum p H is 4.5,the optimum temperature was 60 ℃,the best substrate concentration for 4mmol/L,and michaelis constant（Km） is a tendency for 1.434 mmol/L,Ca^2＋ on the inulase activation function,when the concentration of join tendency for 1.5mmol/L of Ca^2＋,the enzyme activity increased from 0.66 U/m L to 1.09 U/m L,Mn2＋,Cu^2＋ and Mg2＋ internal cutting type inulinase has inhibitory effect,in which inhibition of Cu^2＋,the enzyme activity,down from 0.66 U/m L to 0.095 U/m L. Na^＋,K＋,Zn2＋,Fe2＋,Mg2＋plus the inulin little effects on the enzyme activity of the enzyme; pH in the range of 4.5 - 8.0,the temperature is between 50 - 60 ℃ inulinase activity is relatively stable.

关 键 词：内切型菊粉酶 酶学性质 黑曲霉 底物浓度 米氏常数 

分 类 号：TS201.3[轻工技术与工程—食品科学]