文章摘要
陶静,丁冬各,陈荫,王斌,王加斌,赵玉勤,霍健聪.带鱼高F值寡肽的制备工艺及活性[J].水产学报,2018,42(10):1648~1660
带鱼高F值寡肽的制备工艺及活性
Preparation and bioactivity evaluation of high Fischer ratio oligo-peptide from Trichiurus japonicus
投稿时间:2017-07-13  修订日期:2018-01-21
DOI:10.11964/jfc.20170710902
中文关键词: 带鱼  高F值寡肽  酶解  脱芳工艺  抗疲劳  抗氧化
英文关键词: Trichiurus japonicus  high Fischer ratio oligo-peptide (HFP)  enzymatic hydrolysis  technology on removing aromatic amino acids  anti-fatigue  antioxidant
基金项目:国家自然科学基金(81673349);浙江省公益技术研究项目(LGN18D060002);浙江省自然科学基金(LY17C200011)
作者单位E-mail
陶静 浙江海洋大学食品与医药学院, 浙江 舟山 316022  
丁冬各 浙江海洋大学食品与医药学院, 浙江 舟山 316022  
陈荫 浙江海洋大学食品与医药学院, 浙江 舟山 316022  
王斌 浙江海洋大学食品与医药学院, 浙江 舟山 316022 wangbin4159@hotmail.com 
王加斌 浙江海力生集团有限公司, 浙江 舟山 316022  
赵玉勤 浙江海洋大学食品与医药学院, 浙江 舟山 316022  
霍健聪 浙江海洋大学食品与医药学院, 浙江 舟山 316022  
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中文摘要:
      以带鱼为原料,通过双酶分步水解制备蛋白酶解液,利用活性炭吸附脱除芳香族氨基酸,紫外和氨基酸组成分析测定高F值寡肽(HFP)的F值,并对昆明小鼠体内的活性进行了系统评价。酶解实验结果显示,带鱼蛋白采用胃蛋白酶和风味蛋白酶进行双酶分步水解,胃蛋白酶的最优酶解工艺参数为酶用量3%,料液比1∶5(g/mL),温度35℃,pH 2.0,酶解时间3 h;风味蛋白酶的最优酶解工艺参数为酶用量3%,pH 7.0,温度50℃,酶解时间3 h。活性炭型号筛选和脱芳工艺研究表明,4号粉末活性炭(200目)优于其他4种活性炭,在料液比1∶20(g/mL)、pH 6.0、常温下处理4 h,蛋液的脱芳率最高,F值(OD220/OD280)由脱芳前的5.75增加到脱芳后的30.72。制备的带鱼高F值寡肽经氨基酸组成分析验证,F值(支链氨基酸/芳香族氨基酸)为28.06。活性实验结果显示,与对照组相比,高F值寡肽剂量组昆明小鼠的醉酒时间明显延长,醒酒时间显著缩短,力竭游泳时间明显延长,游泳后的剂量组肝糖原和肌糖原含量增加,血清尿素氮和乳酸含量下降,而乳酸脱氢酶活性则明显提高。另外,剂量组小鼠体内的抗氧化酶SOD、GSH-Px和CAT的活性均显著提高,而丙二醛含量则显著减少。本实验为带鱼高F值寡肽的制备提供技术支持,所制备的高F值寡肽对小鼠具有较好的抗醉酒、醒酒、抗疲劳作用和抗氧化活性。
英文摘要:
      In the experiment, the preparation technology of the high Fischer ratio oligo-peptide (HFP) from Trichiurus japonicus (HFP-T) was studied, and the parameters on two-step sequential enzymolysis and removing aromatic amino acids (AAA) using activated carbon were optimized. The F value of the HFP was confirmed by determination of amino acid components. Taking hydrolysis degree and protein conversion ratio as criterions, binary-enzymes hydrolysis process was optimized and the results were as follows: pepsin was selected as the first enzyme, and its optimal parameters were ratio of enzyme to substrate 3%, ratio of meat to water 1∶5 (g/mL), temperature 35 ℃, pH 2.0 and enzymolysis time 3 h; flavourzyme was selected as the second enzyme, and its optimal parameters were ratio of enzyme to substrate 3%, pH 7.0, temperature 50 ℃ and enzymolysis time 3h. The highest hydrolysis degree and F value (OD220/OD280) of hydrolysate prepared under the optimal parameters were 40.51%±3.18% and 5.75, respectively. In order to raise the F value of the sample by removal of AAA, five kinds of activated carbons including one powdered and four granular activated carbons were chosen to evaluate their removing AAA effects, and the results indicated that No.4 activated carbon (200 mesh) have the best removing AAA effects under the parameters of solid-liquid ratio 1∶20 (g/mL) and pH 6.0 for 4 h at room temperature, and the F value (OD220/OD280) of hydrolysate reached 30.72 from 5.75. The F value (BCAA/AAA) of hydrolysates (removing AAA) measured on Automatic Amino Acid Analyzer was calculated to be 28.06, and the content of Lysine was 22.403 μmol/L. The results of activity tests showed that HFP-T can significantly prolong time of the loss of righting reflex and shorten time to sober up, increase liver and muscle glucogen contents and lower blood urea nitrogen and blood lactate levels after mice have exercised. HFP-T also enhanced the activity of lactic dehydrogenase to scavenge excessive LA for slowing the development of fatigue. In addition, HFP-T increased the activities of superoxide dismutase, catalase and glutathione peroxidase to reduce the reactive oxygen species (ROS) damage in mice. In conclusion, HFP-T possessed good anti-fatigue capacities in mice by inhibiting the oxidative reactions and provided an important basis for developing the swim bladder peptide functional food.
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