抑制α-葡萄糖苷酶活性的啤酒糟多肽的合成与表征
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摘要
采用以Fmoc为保护基团的固相合成法来合成啤酒糟多肽SS-5(序列:Ser-Pro-Asp-Arg-Ser),以高效液相色谱(HPLC)技术对合成肽粗品进行纯化脱盐,得到纯度95%以上的样品。合成肽SS-55对α-葡萄糖苷酶的抑制作用检测结果表明,合成肽SS-5的活性受所含盐离子浓度的影响,低浓度时抑制率较高。而圆二色谱和荧光猝灭试验结果表明,合成肽SS-5与α-葡萄糖苷酶之间存在相互作用。说明可以通过固相合成法快速大量获得对α-糖苷酶具有一定抑制作用的啤酒糟多肽SS-5。
Brewer's grains peptide SS-5(sequence;Ser-Pro-Asp-ArgSer) was synthesized by solid-phase synthesis with Fmoc protect group.Then the crude synthetic peptide was purified and desalted by HPLC,and as a result the purity was over 95%.The inhibitory activity of synthetic peptide SS-5 to α-glucosidase was detected,and the results showed that it was influenced by salt ionic concentration and the lower concentration presented higher mhibihty.Furthermore,the interaction between synthetic peptide SS-5 and α-glucosidase was confirmed by circular dichroism spectrum(CD) and fluorescence quenching assays..Therefore,a fast method of obtaining brewer's grains peptide SS-5 with α-glucosidase inhibitory activity in quantity was proposed in this paper.
Brewer's grains peptide SS-5(sequence;Ser-Pro-Asp-ArgSer) was synthesized by solid-phase synthesis with Fmoc protect group.Then the crude synthetic peptide was purified and desalted by HPLC,and as a result the purity was over 95%.The inhibitory activity of synthetic peptide SS-5 to α-glucosidase was detected,and the results showed that it was influenced by salt ionic concentration and the lower concentration presented higher mhibihty.Furthermore,the interaction between synthetic peptide SS-5 and α-glucosidase was confirmed by circular dichroism spectrum(CD) and fluorescence quenching assays..Therefore,a fast method of obtaining brewer's grains peptide SS-5 with α-glucosidase inhibitory activity in quantity was proposed in this paper.
引文
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[15]欧阳嘉,李鑫,潘宗敏,等.Fmoc(9-芴甲氧羰基)法固相合成胸腺五肽[J].南京工业大学学报,2005,27(3):61-64.
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[19]杨城,管骁,韩飞,等.木犀草素抑制酪氨酸酶活性的分子机制[J].分析测试学报,2015,34(5):532-538.
[2]姜丽燕.2型糖尿病患者大血管并发症与LDL-C的相关性分析[D].郑州:郑州大学,2013:1-2.
[3]屠洁,李前龙.天然产物中α-葡萄糖苷酶抑制剂的筛选研究进展[J].食品研究与开发,2010,31(9):206-210.
[4]李玉萍,白冰,叶军,等.α-葡萄糖苷酶抑制剂的制备和活性研究进展[J].食品科学,2008,29(9):617-620.
[5]BHANDARI M R,JONG-ANURAKKUN N,GAO Hong,KAWABATA J.Alpha-glucosidase and alpha-amylase inhibitory activities of Nepalese medicinal herb Pakhanbhed(Bergeniaciliata Haw.)[J].Food Chemistry,2008,106(1):247-252.
[6]WANG Hui,DU Yang-ji,SONG Hua-can.Alpha-Glucosidase and alpha-amylase inhibitory activities of guava leaves[J].Food Chemistry,2010,123(1):6-13.
[7]KIM J S,HYUN T K,KIM M J.The inhibitory effects of ethanol extracts from sorghum,foxtail millet and proso millet on alpha-glucosidase and alpha-amylase activities[J].Food Chemistry,2011,124(4):1 647-1 651.
[8]CHEN Jing,CHENG Yong-qiang,YAMAKI K,et al.Anti-alpha-glucosidase activity of Chinese traditionally fermented soybean(douchi)[J].Food Chemistry,2007,103(4):1 091-1 096.
[9]YU Zhi-peng,YIN Yong-guang,ZHAO Wen-zhu,et al.Novel peptides derived from egg white protein inhibiting alpha-glucosidase[J].Food Chemistry,2011,129(4):1 376-1 382.
[10]林鸿佳.具有α-糖苷酶抑制作用的啤酒糟多肽的分离及其作用机理研究[D].广州:华南理工大学,2013.
[11]崔洁,李琳,张乔会,等.山杏整仁的酶法制备条件优化及功能评价[J].食品科学,2015,36(19):153-158.
[12]李婷婷,王君伟.口蹄疫合成肽疫苗研究进展[J].动物医学进展,2012,33(4):76-79.
[13]王德芝,周则卫,刘鉴峰,等.心肌肌钙蛋白I(cTnI)合成肽制备、鉴定及抗体研究[J].免疫学杂志,2013,9(29):796-799.
[14]王秋玲,张晶.乙型肝炎病毒PreS1合成肽制备PreS1抗体及其应用[J].现代预防医学,2004,31(5):794-795.
[15]欧阳嘉,李鑫,潘宗敏,等.Fmoc(9-芴甲氧羰基)法固相合成胸腺五肽[J].南京工业大学学报,2005,27(3):61-64.
[16]潘宇,朱学军,牟颖.高效液相色谱法分析纯化合成肽[J].生命科学仪器,2008,9(6):13-18.
[17]OHTA T,SASAKI S,OOHORI T,et al.α-Glucosidase inhibitory activity of a 70%methanol extract from ezoishige(Pelvetiababingtonii de Toni)and its effect on the elevation of blood glucose level in rats[J].Biosci.,Biotechnol.,Biochem.,2002,66(7):1 552-1 554.
[18]DU Zhi-yun,LIU Rong-rong,SHAO Wei-yan,et al.α-Glucosidase inhibition of natural curcuminoids and curcumin analogs[J].Eur.J.Med.Chem.,2006,41(2):213-218.
[19]杨城,管骁,韩飞,等.木犀草素抑制酪氨酸酶活性的分子机制[J].分析测试学报,2015,34(5):532-538.