大孔树脂法纯化黑豆酯酶的工艺优化

Optimization of purification process of black soybean esterase by mac-ro-porous absorbent resin adsorption

中文关键词:  黑豆酯酶  大孔树脂  吸附法  纯化  正交实验

英文关键词:black soybean esterase  macro-porous resin  adsorption method  purification  orthogonal test

中文摘要:

      目的 对采用大孔树脂法纯化黑豆酯酶的工艺进行优化, 确定其最佳工艺条件。方法 首先通过静态吸附和洗脱试验, 以黑豆酯酶的吸附率和洗脱率为指标, 筛选最佳树脂。然后进行动态吸附和洗脱试验, 研究上样稀释倍数、洗脱剂体积分数和洗脱剂体积3因素对洗脱率的影响。最后以黑豆酯酶得率和比活力为指标, 用L16(43)正交实验进行优化。结果 通过筛选发现, D301R树脂吸附率和洗脱率都较高。动态吸附和洗脱实验中获得最佳工艺条件为: 上样稀释倍数为40倍, 洗脱剂体积分数为60%, 洗脱剂用量为20 mL。在此条件下黑豆酯酶得率为 (89.39±1.65) %, 黑豆酯酶比活力(6.585±0.090) U/mg。结论 与粗酶的纯度相比, 优化后的大孔树脂法纯化黑豆酶工艺的提纯倍数达到6.143倍,为黑豆酯酶的提纯和有机磷农药残留的检测提供了科学依据。

英文摘要:

      Objective To optimize the purification process of black soybean esterase by macro-porous absorbent resin adsorption. Methods Firstly, the best resin was chosen by the static adsorption and desorption experiments. Then effects of sample dilution multiple, eluent volume fraction and eluent volume on desorption rate were determined by dynamic adsorption and desorption experiments. Finally, orthogonal experiments of L16 (43) were used to optimize the purification process and yield and specific activities of black bean esterase were tested. Results The D301R resin had higher adsorption and desorption rates than others, and the optimum purification process conditions were as follows: the sample dilution volume was 40 times, eluent volume fraction was 60% and eluent volume was 20 mL. Under these conditions, the yield of black beans esterase by macro-porous resin was (89.39±1.65)%, and the specific activity of black beans esterase was (6.585±0.090) U/mg. Conclusion The yield of black soybean esterase purified with optimized macro-porous absorbent resin was increased by 6.143 times compared with crude black bean esterase. The result is useful to purify and determine the residues of organophosphorus pesticides for black soybean esterase.