食品科学

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响应面优化鱿鱼须脱皮液胶原肽酶解工艺及抗氧化活性

罗春艳1,吴杨阳1,孙海燕1,2,余 辉1,江旭华3,陈小娥1,2,*   

  1. 1.浙江海洋大学食品与医药学院,浙江 舟山 316022;2.浙江省水产品加工技术研究联合重点实验室,浙江 舟山 316022;
    3.浙江富丹旅游食品有限公司,浙江 舟山 316104
  • 出版日期:2016-11-15 发布日期:2016-11-18

Optimization of Enzymatic Hydrolysis of Collagen-Rich Waste Liquid from Squid Tentacle Peeling for Preparing Antioxidant Peptides Using Response Surface Methodology and Their Antioxidant Activities

LUO Chunyan1, WU Yangyang1, SUN Haiyan1,2, YU Hui1, JIANG Xuhua3, CHEN Xiaoe1,2,*   

  1. 1. School of Food and Pharmacy, Zhejiang Ocean University, Zhoushan 316022, China;
    2. Joint Key Laboratory of Zhejiang Province for the Research of Fishery Processing Technology, Zhoushan 316022, China;
    3. Zhejiang Fudan Tourism Food Co. Ltd., Zhoushan 316104, China
  • Online:2016-11-15 Published:2016-11-18

摘要:

目的:研究秘鲁鱿鱼须脱皮液胶原肽酶解工艺及抗氧化活性。方法:以水解度和1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率为指标,利用单因素试验和响应面法对其酶解工艺进行优化,并采用2,2’-联氮-二(3-乙基苯并噻唑啉-6-磺酸)二铵盐(2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt,ABTS)自由基与•OH清除率及亚铁还原能力评价其酶解产物抗氧化活性以及通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)和凝胶排阻色谱分析其分子质量分布。结果:最适酶解工艺为酶解温度50 ℃、初始pH 7.4、底物蛋白质质量分数3.2%、酶解时间3.7 h、酶添加量3 000 U/g。在此条件下,水解度和酶解产物DPPH自由基清除率分别为(37.23±0.08)%和(43.61±0.09)%,对ABTS+•和•OH清除率的IC50分别为0.37 mg/mL和0.41 mg/mL,且表现出较强的亚铁还原能力;SDS-PAGE显示最适酶解工艺下,蛋白质基本被水解为小分子多肽;凝胶排阻色谱分析可知,酶解产物分子质量在1~5 kD之间。结论:酶解产物具备较强的抗氧化活性,可作为抗氧化活性物质基料,充分开发利用。

关键词: 鱿鱼须脱皮液, 酶解, 抗氧化, 水解度, 半抑制浓度

Abstract:

Objective: To investigate the optimum parameters for enzymatic hydrolysis of collagen-rich waste liquid from
squid (Dosidicus gigas) tentacle peeling and evaluate the antioxidant activity of its hydrolysate. Methods: The hydrolysis
parameters were optimized using one-factor-at-a-time method and response surface methodology based on 1,1-diphenyl-2-
picrylhydrazyl (DPPH) radical scavenging activity and degree of hydrolysis (DH). The antioxidant activity of hydrolysate
was evaluated by using scavenging rate of 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt radical
(ABTS+·) and hydroxyl radical scavenging and ferric reducing/antioxidant power (FRAP) assays and the molecular weight
distribution was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration
chromatography. Results: The optimal hydrolysis parameters were determined as follows: hydrolysis temperature, 50 ℃;
substrate (protein) content, 3.2%; hydrolysis time, 3.7 h; enzyme dosage, 3 000 U/g; and initial pH, 7.4, resulting in a DH
value of (37.23 ± 0.08) % and a scavenging rate of DPPH radical of (43.61 ± 0.09) %, respectively. The half inhibitory
concentrations for ABTS+· and hydroxyl radical scavenging ability (IC50) of the hydrolysate obtained were 0.37 and
0.41 mg/mL, respectively. Moreover, the product showed a strong reducing capacity. SDS-PAGE analysis showed that most
collagens in the waste liquid were hydrolyzed into small peptides with molecular weight of 1?5 kD as determined by gel
exclusion chromatography. Conclusion: The hydrolysate derived from waste liquid from squid tentacles peeling has strong antioxidant
activity and could be used as a functional seafood condiment and nutritional supplement for further development and utilization.

Key words: waste liquid from squid tentacle peeling, enzymatic hydrolysis, antioxidant activity, degree of hydrolysis, half inhibitory concentration

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