响应面试验优化绿豆凝集素提取工艺

凝血法检测绿豆凝集素活性，并改进凝集活性检测兔血红细胞处理条件，在单因素试验的基础上设计响应面Box-Behnken试验，优化绿豆凝集素提取工艺条件。结果表明，凝集活性检测兔血红细胞的处理最佳条件为戊二醛体积分数0.15%、25 ℃处理血细胞20 min；胰蛋白酶添加量25 U/mL、25 ℃处理15 min，优化条件下提高了兔血红细胞凝集灵敏度且延长了兔血红细胞保存时间。磷酸盐缓冲溶液为绿豆凝集素最佳浸提溶液，绿豆凝集素优化工艺条件为料液比1∶9.09（g/mL）、NaCl浓度0.3 mol/L、浸提时间4.16 h，在此条件下最大绿豆凝集素比活力预测和验证值分别为134.91 U/mg和139.02 U/mg。

Response Surface Methodology to Optimize the Extraction Process of Mung Bean Lectin

The experiment was designed to use erythrocytes for assaying the agglutinating activity of mung bean lectin,
and to improve the processing conditions for glutaraldehyde- and trypsin-treated rabbit red blood cells. Rabbit red blood
cells (RBC) were fixed with 0.15% (V/V) glutaraldehyde at 25 ℃ for 20 min, and the RBC were treated with trypsin
(25 U/mL) at 25 ℃ for 15 min to increase the assay activity and extend their preservation time. In the investigation, we
found that the most suitable extraction solvent for mung bean lectin was phosphate buffer solution. Based on the results of
single-factor experiments, a Box-Behnken design (BBD) of three variables at three levels each was carried out to obtain
maximum activity of mung bean lectin by response surface analysis. The maximum activity of mung bean lectin was
predicted to be 134.91 U/mg and observed to be 139.02 U/mg under the optimum extraction conditions: solid-to-liquid ratio,
1:9.09 (g/mL); NaCl concentration, 0.3 mol/L; and extraction time, 4.16 h.