Optimization of enzymolysis conditions for degradation of agarose by cellulase
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摘要: 本文以还原糖生成量为指标,研究纤维素酶降解琼脂糖的条件。在单因素实验的基础上,通过Box-Behnken实验设计和响应面分析对琼脂糖酶解条件进行优化,确定4个因子（纤维素酶含量、酶解温度、p H和酶解时间）的适宜水平,建立纤维素酶含量、酶解温度、p H和酶解时间与酶解率之间的回归模型,并对酶解产物进行初步鉴定。最优酶解条件:底物含量0.2%、纤维素酶含量1909.3 U/m L、酶解温度50.0℃、p H4.9、酶解时间24.2 h,该条件下琼脂糖的酶解率为29.58%±0.26%。薄层色谱（TLC）鉴定结果表明酶解主要产物为聚合度26的琼胶低聚糖。Abstract: In this paper, the conditions of cellulose enzymatic degradation of agarose were studied with the yield of reducing sugar as the index. Following single factor experiment, the enzymatic hydrolysis conditions of agarose were optimized by four factors ( cellulase content, enzymolysis temperature, p H and enzymolysis time) Box-Behnken experiment design and response surface methodology ( RSM) , and a regression model was established for predicting the enzymatic hydrolysis yield at different levels of cellulase content, enzymolysis temperature, p H and enzymatic hydrolysis time. In addition, the enzymatic hydrolysate products were preliminary identified. The optimum enzymolysis parameter, including, the substrate content was 0.2%, the cellulase content was 1909.3 U/m L, the enzymolysis temperature was 50.0 ℃, the p H was 4.9, and the enzymolysis time was24.2 h. Under these conditions, the enzymolysis rate of agarose reached 29.58% ± 0.26%. The results of thin layer chromatography ( TLC) showed that the main products were agar oligosaccharides with polymerization degree of 26.
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