超高压辅助酶法制备黑斑蛙皮抗氧化肽及其活性评估  

Extraction of Antioxidant Peptides and Activity Evaluation of Frog(Rana nigromaculata)Skin by Ultra-high Pressure Assisted Enzyme Hydrolysis

作　　者：张喜才[1,2] 李蓉[1] 黄业传 ZHANG Xicai;LI Rong;HUANG Yechuan(School of Biological Engineering,Jingchu University of Technology,Jingmen 448000,China;Institute of Agricultural Biotechnology,Jingchu University of Technology,Jingmen 448000,China)

机构地区：[1]荆楚理工学院生物工程学院,湖北荆门448000 [2]荆楚理工学院农业生物技术研究所,湖北荆门448000

出　　处：《食品工业科技》2025年第4期167-175,共9页Science and Technology of Food Industry

基　　金：湖北省科技厅重点研发计划(2022BCE057);湖北省教育厅科学研究计划资助项目(D20214303);荆楚理工学院重点科学研究项目(ZD202313)。

摘　　要：为提高黑斑蛙加工副产物蛙皮的综合利用价值,对黑斑蛙蛙皮抗氧化肽的提取工艺及性能进行研究。采用超滤方法对黑斑蛙蛙皮的蛋白酶解液进行分离,得到不同分子量范围的小分子肽,通过体外抗氧化试验和秀丽隐杆线虫模型,评价不同分子量范围蛙皮小分子肽的抗氧化性能;以高活性小分子肽得率为目标,采用响应面方法对酶添加量、超高压压力、超高压时间、液料比四个参数优化,确定小分子肽最佳提取工艺。结果表明,1~2000 Da小分子肽具有最强的抗氧化能力,DPPH自由基、ABTS^(+)自由基清除率分别达到了76.93%、59.23%,并显著提高了秀丽隐杆线虫的抗氧化能力;在中性蛋白酶水解的基础上,二次酶解最佳工艺参数为碱性蛋白酶用量10210 U/g,超高压压力315 MPa,压力作用时间9.3 min,液料比16倍,1~2000 Da小分子肽得率为25.26%。超高压辅助双酶水解显著提高了黑斑蛙蛙皮小分子肽的得率,为黑斑蛙副产物的高值化利用提供了参考。In order to improve the comprehensive utilization value of the skin of the frog(Rana nigromaculata),the extraction process of antioxidant peptides of the skin of the frog(Rana nigromaculata)was studied in this experiment and its activity was evaluated.Ultrafiltration was used to separate the protease hydrolysis solution of the skin of the frog(Rana nigromaculata)to obtain small molecule peptides with different molecular weight.The antioxidant properties of frog(Rana nigromaculata)skin small molecule peptides were evaluated in Caenorhabditis elegans model as well as in vitro.Taking the yield of highly active small molecule peptides as the index,the response surface method was utilized to optimize the four parameters containing enzyme addition,ultra-high pressure,the action time,and liquid-to-material ratio to determine the best extraction process for small molecule peptides.Results showed that 1~2000 Da small molecule peptides possessed the optimal antioxidant effect compared to other peptides in the same molecular weight range,the scavenging capacity of DPPH free radicals and ABTS^(+)free radicals reached 76.93%and 59.23%,respectively.The antioxidant capacity of Caenorhabditis elegans was significantly improved.Besides,on the basis of neutral protease hydrolysis,the optimum technology parameters for secondary enzymatic hydrolysis were alkaline protease dosage 10210 U/g,ultra-high pressure pressure 315 MPa,pressure action time 9.3 min,liquid-material ratio 16 times,and the yield of small molecule peptides with 1~2000 Da was 25.26%under above condition.Ultra-high pressure assisted two-enzyme hydrolysis significantly increased the yield of small-molecule peptides from the skin of the frog(Rana nigromaculata).The results of this research provide a reference for the high-value utilization of by-products of the frog(Rana nigromaculata).

关 键 词：超高压 双酶水解 黑斑蛙皮 小分子肽 抗氧化性能 

分 类 号：TS201.1[轻工技术与工程—食品科学]