裙带菜乙醇洗脱组分对α-葡萄糖苷酶活性的影响
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摘要
探讨养殖型褐藻裙带菜水提醇沉上清部分洗脱组分对α-葡萄糖苷酶活性的抑制作用及其反应动力学.采用AB-8大孔吸附树脂柱,依次用去离子水、25%乙醇、50%乙醇、75%乙醇、100%乙醇梯度洗脱裙带菜水提醇沉上清部分,获得相应洗脱组分.采用PNPG法测定α-葡萄糖酶活性,并与阳性对照阿卡波糖作比较.随后,对其中抑制α-葡萄糖苷酶活性较强的组分进行酶抑制动力学分析,推断酶抑制类型.结果表明:相比于裙带菜水提醇沉上清部分与其他洗脱组分,50%乙醇洗脱组分具有较强的α-葡萄糖苷酶抑制活性,半效应浓度IC50为0.77mg/mL,远小于阿卡波糖IC506.63 mg/mL,其酶抑制类型属于混合I型抑制,主要通过与游离酶(E)的亲和来起到抑制作用.因此,裙带菜水提醇沉上清部分50%乙醇洗脱组分具有作为新型α-葡萄糖苷酶抑制剂材料的开发价值.
This study aimed to explore the effects of various elutions from the supernatant from water extract-alcohol precipitation of Undaria pinnatifida onα-glucosidase activity and its reaction kinetics.For various elutions,the supernatant from water extract-alcohol precipitation of Undaria pinnatifida was eluted by the AB-8 macroporous resin with water,25%,50%,75%,and 100% ethanol,respectively.The effects of various fractions onα-glucosidase activity were determined by the PNPG method,and acarbose was used as a positive control.The elution fraction with the strongestα-glucosidase inhibitory activity was then selected for the enzymatic kinetic analysis and identifying its inhibitory type.The results showed that the 50%ethanol elution from the supernatant from water extract-alcohol precipitation of Undaria pinnatifida had strongestα-glucosidase inhibitory activity by comparing with the supernatant and other elution fractions.The IC_(50) value of the 50% ethanol elution was 0.77 mg/mL,which was much less than the IC50 of acarbose with 6.63 mg/mL.Enzyme inhibitory type of the 50% ethanol elution was mixed Ⅰtype inhibition,and its inhibitory effects were mainly through the affinity with free enzyme(E).These results indicated that the 50%ethanol elution from the supernatant from water extract-alcohol precipitation of Undaria pinnatifida should have developmental value to be a source of newα-glucosidase inhibitor.
This study aimed to explore the effects of various elutions from the supernatant from water extract-alcohol precipitation of Undaria pinnatifida onα-glucosidase activity and its reaction kinetics.For various elutions,the supernatant from water extract-alcohol precipitation of Undaria pinnatifida was eluted by the AB-8 macroporous resin with water,25%,50%,75%,and 100% ethanol,respectively.The effects of various fractions onα-glucosidase activity were determined by the PNPG method,and acarbose was used as a positive control.The elution fraction with the strongestα-glucosidase inhibitory activity was then selected for the enzymatic kinetic analysis and identifying its inhibitory type.The results showed that the 50%ethanol elution from the supernatant from water extract-alcohol precipitation of Undaria pinnatifida had strongestα-glucosidase inhibitory activity by comparing with the supernatant and other elution fractions.The IC_(50) value of the 50% ethanol elution was 0.77 mg/mL,which was much less than the IC50 of acarbose with 6.63 mg/mL.Enzyme inhibitory type of the 50% ethanol elution was mixed Ⅰtype inhibition,and its inhibitory effects were mainly through the affinity with free enzyme(E).These results indicated that the 50%ethanol elution from the supernatant from water extract-alcohol precipitation of Undaria pinnatifida should have developmental value to be a source of newα-glucosidase inhibitor.
引文
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[9]陈清西.酶学及其研究技术[M].厦门:厦门大学出版社,2010.
[2]康文艺,张丽,宋艳丽.茜草抑制α-葡萄糖苷酶活性成分研究[J].中国中药杂志,2009,34(9):1104-1107.
[3]李亚美,陈黎.对比伏格列波多糖片与阿卡波糖片治疗2型糖尿病的临床效果[J].中国医药指南,2015,13(20):31-33.
[4]盛占武,王会,郑丽,等.香蕉花提取物中抑制α-葡萄糖苷酶活性成分研究[J].中国食品学报,2014,14(9):68-74.
[5]徐婷,褚江洪,丁萍英,等.葛根素对α-葡萄糖苷酶抑制作用机理的探讨[J].中国生化药物杂志,2015,35(1):43-45.
[6]付小梅,高淑清,孙侠,等.裙带菜的生理药理作用研究进展[J].癌变·畸变·突变,2004,16(4):254-256.
[7]张文竹,刘红兵.裙带菜的化学成分及其生物活性研究进展[J].海洋科学,2009,33(4):72-74.
[8]黄晓冬,蔡琼瑶,王远花,等.海藻铁钉菜抑制α-葡萄糖苷酶活性部位的筛选及其反应动力学研究[J].中国医院药学杂志,2016,36(1):16-20.
[9]陈清西.酶学及其研究技术[M].厦门:厦门大学出版社,2010.