拟青霉β-13-葡聚糖酶同源建模及共价固定化.pdfVIP

252 2013, Vol.34, No.01 食品科学 ※生物工程 β -1,3(4)- 1 1 2 2 华承伟 ，于江傲 ，谢凤珍 ，陈晓静 (1.河南科技学院生命科技学院，河南 新乡 453003 ；2.河南科技学院新科学院，河南 新乡 453003) 摘 要： β 采用同源建模的方法构建拟青霉 -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 m m 学参数。结果表明： (酶粉): (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。 β-1,3(4)-葡聚糖酶；同源建模；共价固定化 Homology Modeling and Covalent Immobilization of β-1,3(4)-Glucanase from Paecilomyces sp. FLH30 1 1 2 2 HUA Cheng-wei ，YU Jiang-ao ，XIE Feng-zhen ，CHEN Xiao-jing (1. School of Life Science and Technology, Henan Institute of Science and Technology, Xinxiang 453003, China； 2. College of Xinke, Henan Institute of Science and Technology, Xinxiang 453003, China) Abstract ：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 chrysosporium 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 ratio1.2:1, temperature 40—45 ℃, and immobilization time 8 h. Under these conditions, the protein bindi