金线鱼鱼皮抗冻蛋白酶法制备工艺优化

发布时间：2018-06-30 03:13

本文选题：蛋白 + 酶　；参考：《农业工程学报》2017年05期

【摘要】：为了探讨金线鱼鱼皮抗冻蛋白的酶法高效制备工艺,该文首先探究不同蛋白酶酶解获得的酶解产物的热滞活性(thermal hysteresis activity,THA)以及不同分子区间的酶解产物的热滞活性。随后以抗冻蛋白得率为指标优化了底物与酶的比例、酶解时间、酶解温度以及p H值,并采用响应面法探究抗冻蛋白的最佳酶解工艺。最后使用差示扫描量热仪(differential scanning calorimeter,DSC)测定验证抗冻蛋白的热滞活性。结果表明:在碱性蛋白酶、中性蛋白酶、木瓜蛋白酶以及复合蛋白酶4种酶的最佳酶解条件下,碱性蛋白酶酶解所获得抗冻蛋白热滞活性最大,选择碱性蛋白酶作为制备金线鱼鱼皮抗冻蛋白的最佳蛋白酶;截留分子量为≥5~10 k Da鱼皮蛋白的抗冻效果最佳,热滞活性达1.20?0.07℃,确定≥5~10 k Da鱼皮蛋白得率作为酶解制备工艺的指标;得到金线鱼鱼皮抗冻蛋白最佳的酶解制备工艺条件为:底物与酶比例为32∶1、酶解时间为147 min、酶解温度为50℃和p H值为9,在此条件下,金线鱼鱼皮抗冻蛋白得率为49.25%±1.34%。相比于牛血清蛋白(bovine serum albumin,BSA),其降低了溶液起始结晶的温度;在最佳的保留温度条件下,其热滞活性为1.2℃。碱性蛋白酶酶解制备鱼皮抗冻蛋白是一种快速有效的方式,酶解获得的抗冻蛋白热滞活性高。该结果为综合发展和利用金线鱼鱼皮提供一定的理论依据。
[Abstract]:In order to explore the enzymatic preparation process of anti freeze protein of fish skin, the thermal hysteresis activity (thermal hysteresis activity, THA) and the thermal hysteresis activity of the hydrolysates of the enzyme hydrolysates obtained from different proteases were first explored. Then the ratio of substrate to enzyme was optimized by the yield of antifreeze protein. Enzyme hydrolysis time, enzymolysis temperature and P H value, and the best enzymatic hydrolysis process of antifreeze protein was investigated by response surface method. Finally, the thermal hysteresis activity of antifreeze protein was tested by differential scanning calorimeter (differential scanning calorimeter, DSC). The results showed that 4 kinds of alkaline protease, neutral protease, papain and compound protease were found. Under the optimum enzymatic hydrolysis conditions, the alkaline protease hydrolysis obtained the maximum thermal hysteresis activity of antifreeze protein. The best protease was selected as the alkaline protease to prepare the antifreeze protein of the skin of the fish skin. The best anti freezing effect of the fish skin protein with the molecular weight of 5~10 K Da was the best, the thermal hysteresis activity was 1.20? 0.07 C, and the yield of the fish skin protein of more than 5~10 K Da was determined. The optimum enzymatic hydrolysis preparation conditions were obtained by enzymatic hydrolysis. The optimum enzymatic hydrolysis conditions were: the ratio of substrate to enzyme was 32: 1, the time of enzymatic hydrolysis was 147 min, the temperature of enzymolysis was 50 and P H was 9. Under this condition, the yield of antifreeze protein of the fish skin was 49.25% + 1.34%. compared to bovine serum albumin, B SA) reduces the temperature of the initial crystallization of the solution, and its thermal hysteresis activity is 1.2 C under the optimum retention temperature. The enzyme hydrolysis of alkaline protease is a fast and effective way to produce the antifreeze protein of the fish skin. The enzyme hydrolyzed antifreeze protein has high thermal hysteresis activity. This result provides a certain theoretical basis for the comprehensive development and utilization of the skin of the fish.
【作者单位】：福建农林大学食品科学学院;
【基金】：福建省发改委产业技术联合创新专项项目(闽财指[2015]779号) 福建农林大学高水平大学建设项目(612014042)
【分类号】：TS254.9

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