提高漆酶稳定性的化学修饰方法的研究  

Study on the chemical modification of Laccase with stability enhancement

作　　者：熊亚红[1] 高敬忠[1] 郑坚鹏[1] 

机构地区：[1]华南农业大学理学院生物材料研究所,广东广州510642

出　　处：《化学研究与应用》2011年第8期985-990,共6页Chemical Research and Application

基　　金：华南农业大学"211工程"重点学科建设项目(2009B010100001)资助;国家自然科学基金(青年科学基金项目)(20906034B0604)资助;广东省科技计划项目(2008B080701011)资助;国家林业局948项目(2011-4-75)资助

摘　　要：天然漆酶在其应用条件下容易失活,因此如何提高漆酶的稳定性显得尤为重要和紧迫。以目前广泛应用的商品化漆酶DeniLite IIS为研究对象,采用邻苯二甲酸酐(PA)、丁二酸酐(SA)及马来酸酐(MA)对漆酶进行化学修饰,根据修饰酶的酶活和热稳定性的变化确定SA为漆酶的最佳修饰剂。采用L9(34)正交设计表研究了磷酸盐缓冲液的pH值、SA浓度及修饰时间对修饰酶的酶活和热稳定性的影响,结果表明三种因素之间不存在各阶交互作用,SA修饰漆酶的最佳条件为:磷酸盐缓冲液的pH 7.5,SA浓度2 mmol.L-1,修饰时间1.0 h。经SA修饰后漆酶的酶活可提高50%、50℃下热处理30 min的热稳定性可提高15%。这为天然漆酶的改性提供了一条良好的途径。Native laccase was applied widely in industry.It was becoming very important and urgent because native laccase was amenable to activity loss under its application conditions.In this paper,a commercial native laccase named DeniLite IIS with wide application was researched.This laccase was modified with different anhydrides,such as phthalic anhydride（PA）,succinic anhydride（SA）and maleic anhydride（MA）.SA was confirmed as a best modification reagent for laccase according to enzymatic activity and stability enhancement of modified laccase.The influences of three factors of phosphate buffer pH and SA concentration and modification time on enzymatic activity and stability were studied by L9（34）orthogonal experiment.The results showed that no interaction existed between these three factors.The optimal conditions for modification were that phosphate buffer pH was 7.5,SA concentration was 2 mmol·L-1 and modification time was 1.0 hour.SA modification of laccase under the optimal conditions caused enzymatic activity increased by 50%and thermal stability at 50 oC for 30min enhanced by 15%.This research offered a suitable method for property reform of native laccase.

关 键 词：漆酶 化学修饰 酶活力 稳定性 

分 类 号：O643.32[理学—物理化学]