采用国产Seplite LX-1000EP树脂对青霉菌柚苷酶进行固定化,以期提高其稳定性和利用效率。将单因素试验和Box-Behnken试验设计结合对影响Seplite LX-1000EP树脂固定化柚苷酶的主要影响因素进行分析、优化。结果显示,最佳固定化条件为:酶液pH 6.5、固定化温度为26.6℃、载体投放量为0.79 g、固定化时间为24 h,此时固定化柚苷酶酶活达到3 949.32 U/g,酶活回收率提高至44.79%,蛋白固定率为98.43%,说明将Seplite LX-1000EP树脂用于柚苷酶的固定化是切实可行的。

In order to improve naringinase catalytic performance,covalent immobilization of naringinase using domestic Seplite LX-1000EP resin was studied. Monofactorial experiment and Box-Behnken design were adopted to optimize naringinase immobilization conditions,then the data was analyzed with SAS software version 8. 0. The optimum immobilization conditions were summed up: pH of immobilization system,immobilization temperature,the ration of carrier addition,and immobilized time were 6. 5,26. 6℃,0. 79 g Seplite LX-1000EP resin per 5 mL enzyme solution,and 24 hours,respectively. Under these conditions,the specific activity of immobilized naringinase reached3949. 32 U /g,with an enzyme recovery of 44. 79%,and the protein binding rate was 98. 43%. The results above indicated that Seplite LX-1000EP resin was a good carrier for naringinase immobilization research. This study can provide a basic work for both the industrial production of naringinase and the practical application of this enzyme.