壳聚糖固化绿色木霉A10产纤维素酶的研究

Chitosan-` Immobillized Cellulase from Trichoderma viride A10

用壳聚糖作为载体、戊二醛作为交联剂来固定化木霉A10产纤维素酶。试验发现交联剂的最佳交联浓度为5%。通过L9(34)正交试验,结果显示用该方案来固定化木霉A10号产纤维素酶是可行的,并且固定化效率最高为69.12%,最佳条件为固定化pH值为8、物料比为1∶0.03、处理时间为12 h。固定化后,酶对热的稳定性明显提高;固定化酶的最适pH比原酶要低,往酸性方向移,且在弱酸性范围内受pH的影响较小;表观米氏常数减小。半衰期为35 d左右。

In this study,chitosan was used as immobile carrier and glutaraldehyde was used as cross-linking reagent.In the Orthogonal design there were three factors and three levels.The factors were pH,the proportion between chitosan and enzyme,the time dealt with glutaraldehyde.According to L9（34） Orthogonal design the optimal test was done.The result showed that the optimal density of crosslinking reagent was 5%.The efficiency was 69.12%;the optimal orthogonal condition was pH 8,the proportion 1：0.03,and the time 12 hours.The analysis to the dynamics of the enzyme showed that the hot stability raised obviously;the optimal pH was lower than before and moved to the acid direction,and affected by pH relatively lightly in the weak acid range;the apparent Michaelis constant reduced,that is to say the affinity to Na-CMC increased.The half-life was about 35 days.