固定化灵芝漆酶脱色氨基黑反应体系优化

Optimization of Decolorization Reaction System of Amino Black 10B Using Immobilized Ganoderma Lucidum Laccase

采用响应面分析法,对固定化灵芝漆酶(Lac)降解氨基黑10B脱色反应体系进行优化分析.首先采用单因素实验分别获取反应体系中6个影响脱色率因素的最佳值;然后通过Plackett-Burman设计法对选取的6个影响因素进行筛选,确定主要影响因素为ABTS浓度和Lac酶量;再用最陡爬坡实验逼近ABTS浓度和Lac酶量2个主要因素的最大响应区域,并通过中心组合设计和响应面分析,确定2个主要影响因素的最佳值.优化后的氨基黑10B脱色200μL反应体系为:pH值4.6、温度30℃、ABTS浓度0.07mmol·L-1、硫酸铜浓度1.75mmol·L-1、Lac酶量6.23mg、初始底物浓度100mg·L-1.该反应体系氨基黑10B脱色率可达60.02%,比单因素法脱色率提高了44.46%.

Response surface methodology（RSM）was applied for optimization of amino black 10 Bdecolouration using immobilized ganoderma lucidum laccase.The optimum value of the six selected impact factors about decolorizing efficiency were measured through single factor experiment.A Plackett-Burman design was applied to screen potential key factors,and the steepest ascent method was adopted to approach the approximate optimal reaction system.Finally,central composite design and response surface methodogy were applied to define more precisely the optical composition of the reaction system.The data revealed ABST concentration and Laccase amount to be the most influential parameters,and the composition of the optimal reaction system was determined to consist of pH 4.6,temperature 30 ℃,copper sulfate concentration 1.75 mmol·L-1,ABTS concentration 0.07mmol·L-1,enzyme 6.23 mg,the initial amino black 10B100mg·L-1.The decolorization ratio of amino black 10 Bin the optimal reaction system was 60.02%,which was 44.46% higher than single factor method.