探究褶牡蛎中金属硫蛋白的提取方法并优化其分离制备工艺。以褶牡蛎为原料，采用Cd2+诱导机体产生MT，然后根据单因素实验、Box-Behnken实验原理，以牡蛎组织与提取液物料比、提取温度、提取pH及缓冲液浓度为影响因素，以金属硫蛋白提取量作为响应值，进行牡蛎金属硫蛋白提取工艺响应面优化分析。结果发现，提取条件对金属硫蛋白提取量影响从大到小依次为：物料比>提取pH值>提取温度=缓冲液浓度，进而获得最佳提取工艺参数为：物料比1:4，提取液pH 9.0，提取温度30 ℃，缓冲液浓度0.25 mol/L，提取时间2 h。在此参数条件下，MT提取量为0.221 mg/g，该结果与模型的预测值基本相符。研究结果可为海洋源金属硫蛋白分离提取以及进一步的活性开发与应用，提供一定的参考与基础。

This study was conducted to investigate the extraction of metallothionein (MT) from oyster (Crassostrea plicatul) and optimize its separation and preparation process. According to the single factor experiment and the Box-Behnken experimental principle, the response surface methodology was performed in this experiment, with the material ratio of the oyster tissue to extract, the extraction temperature, the pH value and buffer concentration were taken as the experimental factors, and the extraction capacity of MT was taken as the response value to perform optimization analysis for the extraction process of metallothionein from oyster (Crassostrea plicatul) based on response surface methodology. The results showed that the effects of extraction conditions on the amount of metallothionein extraction descending order were as follows: the ratio of oyster tissue to extraction buffer > pH > extraction temperature = buffer concentration, and then the optimum extraction process parameters were as follows: the material ratio of was 1:4, the value of pH was 9.0, the extraction temperature was 30℃, the concentration of buffer was 0.25 mol/L, and the extraction time was 2 h. Under these conditions, the obtained MT was 0.221 mg/g, which were almost consistent with the predicted data of the model. These results could provide a reference and theoretical basis for the separation and extration of marine metallothionein in addition to its further exploitation and application of the activity.

中国博士后基金项目（2016M590663、2017T100518）；浙江省公益性项目（201632080、201632081）；浙江海洋大学大学生科技创新项目