食品科学 ›› 2017, Vol. 38 ›› Issue (22): 246-255.doi: 10.7506/spkx1002-6630-201722037

• 工艺技术 • 上一篇    下一篇

响应面优化酶法提取虎斑乌贼肌肉多糖的工艺及抗氧化活性测定

孙玉林,文菁,赵娟,田丽,李锐,陈道海,   

  1. (1.岭南师范学院生命科学与技术学院,广东?湛江 524048;2.岭南师范学院环北部湾海洋药用动物保护与利用研究所,广东?湛江 524048)
  • 出版日期:2017-11-25 发布日期:2017-11-03
  • 基金资助:
    广东省科技厅项目(2014B040404071;2015A070708013;2015A030302089); 广东省省部产学研合作专题项目(2013B090500036);广东省自然科学基金项目(2015A030310406;2017A030307029); 广东省高等教育“创新强校工程”项目(2016KTSCX081); 湛江市财政资金科技专项竞争性分配项目(2015A06008;2015A03017;2014A03011); 岭南师范学院博士启动项目(ZL1313);岭南师范学院自然科学研究合作项目(HL1402)

Optimization of Enzymatic Extraction and Antioxidant Activity of Polysaccharides from the Muscle of Sepia pharaonis Using Response Surface Methodology

SUN Yulin, WEN Jing, ZHAO Juan, TIAN Li, LI Rui, CHEN Daohai,   

  1. (1. School of Life Science and Technology, Lingnan Normal University, Zhanjiang 524048, China; 2. Round Beibu Gulf Institute for the Protection and Utilization of Marine Animals in Medicine, Lingnan Normal University, Zhanjiang 524048, China)
  • Online:2017-11-25 Published:2017-11-03

摘要: 研究胰蛋白酶提取虎斑乌贼肌肉多糖的工艺及体外抗氧化活性,并与水提法和碱提法的多糖进行比较。在单因素试验基础上以响应面法优化酶法提取多糖的工艺,结果显示最佳工艺条件为:酶解温度55?℃、料液比1∶40(g/mL)、酶解时间4?h、加酶量2.5%,此时虎斑乌贼肌肉多糖得率为4.15%,优于水提法和碱提法的多糖得率。体外抗氧化活性研究表明酶法提取的多糖比碱提和水提的多糖具有更好的抗氧化活性,当多糖质量浓度为4?mg/mL时,水提、碱提和酶提虎斑乌贼肌肉多糖对·OH的清除率分别为50.15%、55.14%和89.47%,其IC50值分别为4.51、3.56?mg/mL和0.82?mg/mL;对DPPH自由基的清除率分别为28.89%、31.48%和40.80%,其IC50值分别为16.66、15.43?mg/mL和11.50?mg/mL;对O2-·的清除率分别为75.43%、81.63%和97.00%,其IC50值分别为1.15、0.69?mg/mL和0.29?mg/mL;还原力大小分别为0.134、0.156和0.193。综合分析表明,酶提法得到的多糖得率较高且具有较好的体外抗氧化活性。

关键词: 虎斑乌贼, 多糖, 酶法提取, 响应面, 抗氧化活性

Abstract: The trypsin-assisted extraction of polysaccharides from the muscle of cuttlefish (Sepia pharaonis) was optimized using one-factor-at-a-time method and response surface methodology. The in vitro antioxidant activity of the obtained polysaccharides was assessed and compared with that of those obtained by water extraction and alkali extraction. The optimum extraction conditions were determined as follows: hydrolysis temperature, 55 ℃; solid-to-water ratio, 1:40 (g/mL); hydrolysis time, 4 h; and trysin dosage, 2.5%. Under these conditions, the maximum polysaccharide yield of 4.15% was obtained, which was higher than those extracted with water and alkali. The antioxidant tests in vitro showed that the enzymatically extracted polysaccharide displayed higher antioxidant activity than the water- and alkali-extracted ones. At a concentration of 4 mg/mL, the percentage scavenging of hydroxyl radical by the polysaccharides extracted with water, alkali and trypsin were 50.15%, 55.14% and 89.47%, with median inhibition concentration (IC50) values of 4.51, 3.56 and 0.82 mg/mL, respectively; the percentage scavenging of DPPH radical were 28.89%, 31.48% and 40.80% with IC50 values of 16.66, 15.43 and 11.50 mg/mL, respectively; and the percentage scavenging of superoxide anion radical were 75.43%, 81.63% and 97.00% with IC50 values of 1.15, 0.69 and 0.29 mg/mL, respectively. The reducing capacity values of the three polysaccharides were 0.134, 0.156 and 0.193, respectively. Collectively, it was shown that the yield of enzymatically extracted polysaccharides was higher and the polysaccharides had higher antioxidant activity in vitro. The polysaccharides from the muscle of S. pharaonis have the potential to be a new non-hazardous and natural antioxidant.

Key words: Sepia pharaonis, polysaccharides, enzymatic extraction, response surface methodology, antioxidant activity

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