Optimization of Extraction Technology of Metallothionein in Arca subcrenata

The study was aimed to establish an efficient extraction technology of metallothionein. The present experiment was conducted to investigate the extraction of metallothionein (MT) from Arca subcrenata by Zn²⁺ stimulated. The changes of MT content before and after induction were compared. To explore the extraction method of metallothionein from Arca subcrenata and optimize the extraction factors, Tris-HCl buffer was selected as extractant. On the basis of single factor experiments, the factors that had great influence on MT extraction content were selected, including pH of extraction buffer, concentration of buffer, ratio of material to liquid and extraction temperature. Based on Box-Behnken experiment principle, response surface optimization analysis of metallothionein extraction process was carried out on the basis of four factors and three levels with MT extraction content as response value.The results showed that the best optimum extraction conditions of MT in Arca subcrenata were as follows:The ratio of extraction buffer to Arca subcrenata tissue was 6:1 mL/g, the concentration of extractant was 0.18 mol/L, the pH of extracti buffer was 8.45, and the heating temperature was 35℃. Under these conditions, the obtained MT was 0.695 mg/g in the actual experiment which were almost accorded with the predicted data. Therefore, this study established an efficient extraction process of MT from Arca subcrenata.