贯筋藤酶解核桃分离蛋白及其体内抗疲劳作用  

Enzymatic Cleavage of Walnut Isolate Proteins by Dregea sinensis Hemsl.Protease and Its Anti-fatigue Effect In Vivo

作　　者：黄思 张霞[1] 牟泓羽 吴宽 马志星 凌云 赵存朝 HUANG Si;ZHANG Xia;MU Hongyu;WU Kuan;MA Zhixing;LING Yun;ZHAO Cunzhao(Yunnan Agricultural University,Kunming 650201,China;Kunming Biomanufacturing Research Institute Co.,Ltd.,Kunming 650201,China;Yunnan Highland Characteristic Agricultural Industry Research Institute,Kunming 650201,China;Yunnan Specialty Resource Food Bio-Manufacturing Engineering Research Center,Kunming 650201,China)

机构地区：[1]云南农业大学,云南昆明650201 [2]昆明生物制造研究院有限公司,云南昆明650201 [3]云南省高原特色农业产业研究院,云南昆明650201 [4]云南省特色资源食品生物制造工程研究中心,云南昆明650201

出　　处：《食品工业科技》2024年第22期305-313,共9页Science and Technology of Food Industry

基　　金：云南省教育厅科学研究基金项目(2023Y1010);云南省省市一体化专项(202302AN360002);高层次科技人才及创新团队选拔专项(202305AS350025);云南省凤庆县核桃产业科技特派团(202204BI090012)。

摘　　要：为了研究核桃蛋白酶解物的抗疲劳作用,为其功能性产品的开发和利用提供科学依据。本文以超临界萃取的核桃粕为原料,提取核桃分离蛋白(WPI),使用贯筋藤蛋白酶酶解WPI,以水解度为指标,研究最佳酶解条件制备核桃肽,并研究不同分子量的核桃蛋白酶解物对小鼠运动性疲劳的缓解作用。将小鼠随机分为五组,每日经口灌胃受试样品,干预周期为一个月,最后进行负重及力竭游泳实验,测定血清尿素氮(BUN)、乳酸(LA)、肝糖原(HG)、肌糖原(MG)、谷胱甘肽过氧化氢酶(GSH-Px)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量。SDS-PAGE凝胶电泳表明贯筋藤蛋白酶对WPI有明显的降解作用;单因素实验结合响应面试验表明,当料液比为9:100、pH9.6、酶添加质量9.4%、酶解时间180 min、酶解温度63℃时,水解度最高达30.1%;HE染色显示,使用贯筋藤蛋白酶提取的酶解物对小鼠无任何毒副作用;与其他组相比,分子量小于3000 Da的核桃蛋白酶解物干预小鼠的LA和BUN含量有所降低,HG和MG含量提高,延长负重游泳时间较为显著(P<0.05),核桃蛋白酶解物干预的各组小鼠血清中SOD酶、GSH-Px酶活性显著提高(P<0.05)。表明核桃蛋白酶解物可以缓解小鼠运动性疲劳,而小分子量的核桃蛋白酶解物效果更显著,这可能是通过调节体内抗氧化活性实现的。To study the anti-fatigue effect of walnut proteolytic enzymes and to provide scientific basis for the development and utilization of its functional products.Walnut isolated protein(WPI)was extracted from supercritically extracted walnut meal,and the WPI was enzymatically digested by Dregea sinensis Hemsl.protease.The hydrolysis degree(DH)was used as an index to study the optimal conditions for the preparation of walnut peptides,and the effects of different molecular weights of walnut proteins on the alleviation of exercise fatigue in mice were also investigated.The mice were randomly divided into five groups,and the test samples were orally gavaged daily for one month.Finally,weight-bearing and exhaustion swimming experiments were performed to determine the serum urea nitrogen(BUN),lactic acid(LA),hepatic glycogen(HG),myo-glycogen(MG),glutathione peroxidase(GSH-Px),malondialdehyde(MDA),and superoxide dismutase(SOD)contents.SDS-PAGE gel electrophoresis showed that the WPI was significantly degraded by Dregea sinensis Hemsl.Protease.The one-way combined response surface experiment showed that the DH was up to 30.1%when the material-liquid ratio was 9:100,pH9.6,enzyme addition quality 9.4%,enzyme digestion time 180 min,and enzyme digestion temperature 63℃.HE staining showed that the enzyme extracted by the use of Dregea sinensis Hemsl.Protease did not have any toxicity side effects on mice.Compared with other groups,the enzymatic digests with molecular weight less than 3000 Da showed a decrease in LA and BUN content,an increase in HG and MG content,and a more significant prolongation of weight-bearing swimming time(P<0.05),and a significant increase in the activities of SOD enzyme and GSH-Px enzyme in serum of mice in each group in the sample(P<0.05).It indicated that walnut proteolytic digests could alleviate exercise fatigue in mice,and the effect of small molecular weight walnut proteolytic digests was more significant,which might be realized by regulating antioxidant activity in vivo.

关 键 词：核桃蛋白 贯筋藤蛋白酶 SDS-PAGE 运动性疲劳 负重游泳 

分 类 号：R151.2[医药卫生—营养与食品卫生学]