响应面法优化微波辅助酶解合浦珠母贝蛋白工艺

研究微波对酶解合浦珠母贝蛋白的影响，以水游离氨态氮含量和抗氧化活性为指标，选择较好的作用酶，通过单因素试验确定料液比、加酶量（E/S）、微波温度、微波功率及时间和后续水浴时间等因素水平，以水解度和DPPH自由基清除率为响应值，响应面法优化酶解合浦珠母贝蛋白工艺条件。结果表明，微波辅助蛋白酶酶解合浦珠母贝蛋白工艺条件为微波温度58 ℃、微波功率300 W、微波时间17 min、加酶量5 000 U/g，后续在58 ℃条件下再水浴1.5 h。预测响应值为0.224 4，水解度达到26.15%，验证实验证明与响应优化模型预测值误差不大，二次多元拟合度较好。

Optimization of Conditions for Microwave-Assisted Enzymolysis of Pinctada martensii Muscle Proteins Using Response Surface Methodology

The microwave-assisted enzymatic hydrolysis of Pinctada martensii muscle proteins was optimized by response
surface methodology. Alcalase 2.4L was chosen as the best enzyme to hydrolyze Pinctada martensii muscle proteins based
on free ammonia nitrogen content and DPPH radical scavenging activity of hydrolsates. The levels of operating parameters
such as substrate concentration, E/S ratio, solvent temperature, microwave power, radiation time and water bath heating
time were established by single-factor experiments for response surface Box-Behnken design. Based on the mathematical
product of degree of hydrolysis (DH) and DPPH radical scavenging rate, the optimal hydrolysis conditions were determined
as a solvent temperature of 58 ℃, microwave treatment at 300 W for 17 min after addition of 5 000 U/g Alcalase 2.4L, and
subsequent water bath heating at 58 ℃ for 1.5 h. Under these conditions, the maximum predicted DH was 26.15%, which
was multiplied by DPPH radical scavenging rate to obtain 0.224 4. The small difference from the actual values observed in
verification experiments suggested a high degree of fitting.