摘要
采用同源建模的方法构建拟青霉β-1,3(4)-葡聚糖酶的三维结构。通过对其活性位点及表面氨基酸残基侧链的分析,利用氨基载体Sepabeads EC-HA共价固定化葡聚糖酶,优化固定化条件,比较固定化酶与游离酶的酶学参数。结果表明:m(酶粉):m(载体)=1.2:1、温度40～45℃、固定化时间8h,固定化效果最好。蛋白结合率可达91.7%,酶活回收率达87.6%,固定化酶最适温度、热稳定性、pH值稳定性和批次使用稳定性均得到明显提高。
The three-dimensional structure of β-1,3（4）-glucanase from Paecilomyces sp. FLH30 was constructed by means of homology modeling using the crystal structure of endo-β-1,3（4）-glucanase from Phanerochaete chrysosporiurn as a template, and its active site and side chains of surface amino acid residues were analyzed. Sepabeads EC-HA as a carrier of amino groups was used for the covalent immobilization of this enzyme and immobilization conditions were optimized. Meanwhile, enzymatic characteristics of free and immobilized β-1,3（4）-glucanase were compared. The best immobilization results were obtained under the conditions： enzyme/carrier mass ratiol.2：1, temperature 40-45 ℃, and immobilization time 8 h. Under these conditions, the protein binding rate was 91.7% and the activity recovery was 87.6%. The optimum temperature, thermal stability, pH stability and operational stability of immobilized glucanase were all improved when compared to free glucanase.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第1期252-256,共5页
Food Science
