响应面法优化酶法提取蜜环菌多肽及其抗疲劳活性

以蜜环菌子实体为原料,采用酶解法制备蜜环菌多肽。在单因素实验的基础上,根据Box-Behnken中心组合实验设计原理,以水解度为响应值,研究底物浓度、加酶量、酶解时间、酶解温度对响应值的影响。通过游泳实验建立小鼠疲劳模型,考察蜜环菌多肽对小鼠游泳时间、肝糖原、肌糖原、尿素氮（BUN）、乳酸（LA）、乳酸脱氢酶（LDH）等指标的影响。结果表明:酶解法提取蜜环菌多肽的最佳工艺为:底物浓度7.4%,加酶量2%,酶解温度51℃,酶解时间4.6 h,水解度为35.61%,与理论值接近。以中剂量组为例,相对于空白组,蜜环菌多肽可以使小鼠肝糖原和肌糖原的含量分别提高109.01%和58.33%,显著降低代谢产物尿素氮水平36.99%、乳酸水平30.33%,乳酸脱氢酶的活性提高9%,以上数值p<0.05。综上蜜环菌多肽具有提高抗疲劳能力的作用。 

Optimization of enzymatic hydrolysis of Armillaria mellea Vahl ex Fr peptides by response surface methodology and its anti-fatigue ability

Armillaria mellea Vahl ex Fr was obtained by enzymatic hydrolysis with Armillaria mellea Vahl ex Fr powder as raw material.The preparation process of walnut peptide was optimized on foundation of single factor experiment, according to Boxbehnken central composition design principle, the method of response surface analysis was adopted with the degree of hydrolysis as the response value.The effect of concentration of substrate, enzyme dosage, time and temperature on the degree of hydrolysis were studied in this paper.Fatigue model was established by swimming test.Effects of Armillaria mellea Vahl ex Fr Peptides on swimming time, liver glycogen, muscle glycogen, serum urea nitrogen, lactic acid, LDH were determined.The rusults showed that the optimum preparation conditions were obtained as follows: concentration of substrate 7.4%, enzyme dosage 2%, time 4.6 h and temperature 51 ℃, the degree of hydrolysis was 35.61%, which was close to the theoretical value. Take the middle dose group as an example, compared to the blank group, the Armillaria mellea Vahl ex Fr Peptides increased content of liver glycogen and muscel glycogen 109.01% and 58.33%, reduced metabolic product of LA and BUN levels 36.99% and 30.33%, increased LDH activities 9%, their differences were significant ( p < 0.05) . The Armillaria mellea Vahl ex Fr Peptides has the function of anti-fatigue obviously.