生物学杂志

生物学杂志 ›› 2022, Vol. 39 ›› Issue (1): 104-.doi: 10. 3969/ j. issn. 2095-1736. 2022. 01. 104

• 技术方法 • 上一篇    下一篇

木瓜蛋白酶法制备抗氧化活性壳寡糖的工艺优化

  

  1. 广东海洋大学食品科技学院广东省水产品加工与安全重点实验室, 湛江524088
  • 出版日期:2022-02-18 发布日期:2022-02-15
  • 通讯作者: 孙力军,教授,研究方向为水产品质量与安全,E-mail:dfsun01@126.com;王雅玲,教授,研究方向为水产品质量与安全,E-mail:wangylchina@163.com
  • 作者简介:黄晓月,硕士研究生,研究方向为水产品质量与安全,E-mail:15819918379@163.com;毕思远,博士,研究方向为水产品质量与安全,E-mail:bisiyuan0129@126.com;黄晓月和毕思远为共同第一作者
  • 基金资助:
    广东省现代农业产业技术体系创新团队建设专项资金项目(No. 2020KJ149;No. 2020KJ151);国家自然科学基金面上项目(31871898);2020 年度广东省普通高校特色创新项目(2020KQNCX025);2020 年度湛江市科技发展专项资金竞争性分配项目(200915134541577);湛江市科技计划项目(No.2020A03009;2020A01011)

Process optimization of preparation of antioxidant chitooligosaccharides by papain

  1. Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety,College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China
  • Online:2022-02-18 Published:2022-02-15

PDF (PC)

50

摘要：

为明确高抗氧化活性壳寡糖制备的木瓜蛋白酶法工艺,以壳聚糖为原料,酶解产物的DPPH 自由基清除率和总还原力为检测指标,使用单因素试验探究酶底比、酶解温度、pH 值、酶解时间对酶解产物抗氧化活性的影响,确定最佳酶解条件为酶底比20%、酶解温度50. 00 ℃、pH 4. 80。在单因素试验的基础上,对酶解温度、酶解时间和pH值进行Box-Behnken 响应面设计并优化得到最佳酶解条件为酶解温度47. 74 ℃、酶解pH 4. 81、酶解时间1 h,预测壳聚糖酶解物的DPPH 自由基清除率为65. 10%,总还原力为0. 323。在酶解温度48. 00 ℃、酶解pH4. 80、酶解时间1 h条件下重复试验得壳聚糖酶解物的DPPH 自由基清除率为65. 43%±0. 71%,IC50 为1. 86 mg/ mL±0. 10 mg/ mL;总还原力为0. 328±0. 010,相当于39. 44 μg/ mL±2. 94 μg/ mL 抗坏血酸的还原能力,该壳寡糖的平均分子质量为1 603 u,产率为45. 07%±3. 52%,为抗氧化活性壳寡糖的研制提供理论参考。


关键词: 木瓜蛋白酶, 壳聚糖, 壳寡糖, DPPH 自由基清除率, 总还原力

Abstract: In order to clarify the papain process for the preparation of chitooligosaccharides with high antioxidant activity, chitosan was used as raw material, the DPPH radical scavenging rate and total reducing power of the hydrolysates were used as detection indexes.The single factor experiment was used to explore the effects of enzyme substrate ratio, temperature, pH value and time on the antioxidant activity of the hydrolysates. The optimal conditions were determined as enzyme substrate ratio of 20%, temperature of 50. 00 ℃ and pH value of 4. 80. On the basis of single factor experiment, box Behnken response surface method was used to design and optimize the hydrolysis temperature, time and pH value. The optimal hydrolysis conditions were obtained as follows: temperature of 47. 74 ℃,pH of 4. 81 and time of 1 h. Under these conditions, the DPPH radical scavenging rate of chitosan hydrolysate was 65. 10% and the total reducing power was 0. 323. The experiment was repeated under the conditions of enzymolysis temperature of 48. 00 ℃, enzymolysis pH of 4. 80 and enzymolysis time of 1 h, the result showed that the DPPH radical scavenging rate of chitosan hydrolysates was 65. 43%±0. 71%, IC50 was 1. 86 mg / mL±0. 10 mg / mL, and the total reducing power was 0. 328 ± 0. 01, which was equivalent to the reducingpower of 39. 44 μg/ mL±2. 94 μg/ mL ascorbic acid. The average molecular weight of this chitooligosaccharide was 1 603 u and the yield of this chitooligosaccharide was 45. 07% ± 3. 52%. The result can provide a theoretical reference for the development of chitooligosaccharides with antioxidant activity.

Key words: papain, chitosan, chitooligosaccharides, DPPH radical scavenging rate, total reducing power

中图分类号: