长足大竹象外切葡聚糖酶活性最适反应条件的初步探究  

The optimal reaction conditions of exo-glucanase from Cyrtotrachelus buqueti(Coleoptera:Curculionidae)

作　　者：唐昊 王昌吉 王明珺 甘晓凤 张鑫月 罗朝兵 TANG Hao;WANG Changji;WANG Mingjun;GAN Xiaofeng;ZHANG Xinyue;LUO Chaobing(College of Sciences,Leshan Normal University,Leshan 614000,China)

机构地区：[1]乐山师范学院生命科学学院,四川乐山614000

出　　处：《甘肃农业大学学报》2021年第3期95-102,共8页Journal of Gansu Agricultural University

基　　金：四川省科技计划项目(2019YGF0139);竹类病虫防控与资源开发四川省重点实验室开放项目(ZLKF1902).

摘　　要：【目的】为客观评价昆虫消化道外切葡聚糖酶的反应条件,丰富纤维素降解酶资源.【方法】通过单因素试验和正交优化试验设计,采用DNS法探究长足大竹象(Cyrtotrachelus buqueti)消化道外切葡聚糖酶活性的最适反应条件.【结果】长足大竹象幼虫外切葡聚糖酶最适反应条件为:反应温度40℃,pH 4.8,底物反应浓度为0.5%;成虫(雌):反应温度30℃,pH 5.2,底物反应浓度为2%;成虫(雄):反应温度为30℃,pH 4.8,底物反应浓度2%.外切葡聚糖酶在经优化后的最佳反应条件下,酶比活力为成虫雌28.26 U/mg、成虫雄26.42 U/mg、幼虫21.50 U/mg.在最佳单因素及正交反应条件下,幼虫酶比活力显著低于成虫.【结论】长足大竹象外切葡聚糖酶活性最适反应条件的研究可以更加深入的了解长足大竹象消化道的纤维素降解酶系统和为进一步解决竹材木质纤维素的生物降解过程的某些技术难题提供部分参考信息.【Objective】In order to evaluate the enzyme reaction conditions of exo-glucanase in intestine of the Cyrtotrachelus buqueti and to enrich the cellulolytic enzyme resource.【Method】The single factor experiment and orthogonal optimized test for investigating the optimal reaction conditions of exo-glucanase activity of the insect were designed and performed by DNS method.【Result】The optimal reaction conditions of exo-glucanase activity in C.buqueti larvae with reaction temperature of 40℃,pH 4.8,substrate reaction concentration of 0.5%;in female adults with reaction temperature of 30℃,pH 5.2 and substrate reaction concentration of 2%;and in male adults was 30℃reaction temperature,pH 4.8,and 2%substrate concentration.The exo-glucanase activities under the optimal reaction conditions of orthogonal optimization test were 28.26 U/mg(female adult),26.42 U/mg(male adult)and 21.50 U/mg(larvae).The enzyme activity of larvae was significantly lower than adults under the optimal reaction conditions of both single factor test and orthogonal optimization test.【Conclusion】The study of the optimal reaction conditions for exo-glucanase activity of C.buqueti could help to understand the cellulolytic enzyme system of the digestive tract of C.buqueti and to provide some reference information for further solving the certain technical problems of the biodegradation process of bamboo lignocellulose.

关 键 词：长足大竹象 外切葡聚糖酶 正交优化试验 反应温度 PH 底物浓度 

分 类 号：Q814.9[生物学—生物工程]