条斑紫菜多糖对α-淀粉酶的抑制效果
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摘要
目的分析条斑紫菜多糖对α-淀粉酶的抑制效果。方法从条斑紫菜中提取多糖α-淀粉酶抑制剂(α-amylase inhibitor,α-AI),分析其在不同浓度(2.5~30 mg/m L)下,经热(30~90℃,20 min)、酸(p H 2.0~7.0,室温,30 min)、碱(p H 7.0~10.0,室温,30 min)、蛋白酶(分别为100 U/g的风味蛋白酶、中性蛋白酶和碱性蛋白酶,50℃,2 h)以及模拟胃肠道环境(胃处理:p H 2.0,100 U/g胃蛋白酶;肠道处理:p H 8.2,100 U/g胰蛋白酶)处理后对α-AI活性的影响,探讨α-AI的稳定性及其对淀粉酶的抑制效果。结果条斑紫菜多糖α-AI在多糖浓度为2.5~30 mg/m L之间对α-淀粉酶的抑制呈剂量依赖关系,浓度为30 mg/m L具有较高的抑制率。经不同条件处理,α-AI活性无显著变化(P>0.05),仍具有良好的淀粉酶抑制活性,且其活性在胃液和肠液中稳定,可较好地发挥生理作用。结论条斑紫菜多糖α-AI在控制血糖稳定方面具有较强的应用价值,可作为2型糖尿病患者潜在的功能性食品。
Objective To analyze the inhibitory effect of polysaccharide from Porphyra yezoensis on α-amylase.Methods Polysaccharide α-amylase inhibitor(α-AI)was extracted from P. yezoensis and analyzed for activity change at various concentrations(2. 5 ~ 30 mg/m L)as well as after treatment by heat(30 ~ 90 ℃,20 min),acid(p H 2. 0 ~ 7. 0,room temperature,30 min),base(p H 7. 0 ~ 10. 0,room temperature,30 min)and protease(100 U/g flavourzyme,neutral protease and alkaline protease respectively,50 ℃,2 h)and in mimic gastrointestinal environment(100 U/g pepsin for gastric treatment,p H 8. 2,100 U/g trypsin for enteric treatment),and evaluated for stability and inhibitory effect on amylase. Results The α-AI at a concentration of 2. 5 ~ 30 mg/m L showed a dose-dependent inhibitory effect on α-amylase.The activity of α-AI showed no significant change after treatment under various conditions(P > 0. 05) and was stable in gastric and enteric fluids. Conclusion The α-AI from P. yezoensis may be used for control of blood glucose,which may be developed as a potentially functional food for patients with type 2 diabetes.
Objective To analyze the inhibitory effect of polysaccharide from Porphyra yezoensis on α-amylase.Methods Polysaccharide α-amylase inhibitor(α-AI)was extracted from P. yezoensis and analyzed for activity change at various concentrations(2. 5 ~ 30 mg/m L)as well as after treatment by heat(30 ~ 90 ℃,20 min),acid(p H 2. 0 ~ 7. 0,room temperature,30 min),base(p H 7. 0 ~ 10. 0,room temperature,30 min)and protease(100 U/g flavourzyme,neutral protease and alkaline protease respectively,50 ℃,2 h)and in mimic gastrointestinal environment(100 U/g pepsin for gastric treatment,p H 8. 2,100 U/g trypsin for enteric treatment),and evaluated for stability and inhibitory effect on amylase. Results The α-AI at a concentration of 2. 5 ~ 30 mg/m L showed a dose-dependent inhibitory effect on α-amylase.The activity of α-AI showed no significant change after treatment under various conditions(P > 0. 05) and was stable in gastric and enteric fluids. Conclusion The α-AI from P. yezoensis may be used for control of blood glucose,which may be developed as a potentially functional food for patients with type 2 diabetes.
引文
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[16]RANG Y F.Preparation of white kidney beanα-amylase inhibitor and its application in instant porridge with low GI[D].Wuxi:Jiangnan University,2016.(in Chinese)让一峰.白芸豆α-淀粉酶抑制剂的制备及其在低GI方便粥中的应用[D].江苏无锡:江南大学,2016.
[17]DING G Y,HUANG L G,AN B,et al.Study on LJPS's effect on decreasing hypoglycemic and hypolipidemic[J].Health Med Res Prac,2010,7(4):10-11.(in Chinese)丁国玉,黄连光,安斌,等.海带多糖降血脂、血糖研究[J].保健医学研究与实践,2010,7(4):10-11.
[18]RANG Y F,ZHAO W,YANG R J,et al.Study on activity changes ofα-amylase inhibitor from white kidney beans during processing and digestion[J].Sci Technol of Food Indust,2015,36(17):53-57.(in Chinese)让一峰,赵伟,杨瑞金,等.白芸豆α-淀粉酶抑制剂在加工和消化过程中的活性变化研究[J].食品工业科技,2015,36(17):53-57.
[19]SU Z H.Research on screening marine microorganism of secretingα-amylase inhibitors and purifying the active ingredient[J].Nanning:Guangxi Normal University,2013.(in Chinese)苏照环.海洋生境分泌α-淀粉酶抑制剂微生物的筛选及活性成分纯化的研究[D].广西南宁:广西师范大学,2013.
[20]YU L G.Ultrafiltration,purification and characterization of polysaccharides from Porphyra haitanensis[D].Xiamen:Jimei University,2012.(in Chinese)于立国.坛紫菜多糖超滤纯化与理化性质的研究[D].福建厦门:集美大学,2012.
[21]GONDA R K,TOMODA M,KANARI M,et al.Constituents of the seed of Malva verticillata.III.:Characterization of the major pectic peptidoglycan and oligosaccharides[J].Chem Pharm Bull,1988,36(8):2790-2795.
[22]ZHU X J.Research on extraction,purification and in vitro antitumor activity of polysaccharides from Porphyra yezoensis[D].Nanjing:Nanjing Agricultural College,2008.(in Chinese)朱晓君.条斑紫菜多糖的提取纯化及其体外抗肿瘤活性研究[D].江苏南京:南京农业大学,2008.
[23]LONG S H.The hypoglycemic effect and mechanism of luminaria japonica polysaccharide in type 2 diabetes mellitus model mice[D].Qingdao:Qingdao University,2013.(in Chinese)龙少华.海带多糖在2型糖尿病小鼠中的降糖作用和机制[D].山东青岛:青岛大学,2013.
[24]ZHAO R.Purification and nature research on lose weight medicineα-amylase inhibitor from white kidney beans[D].Xi'an:Northwest University,2010.(in Chinese)赵蓉.白芸豆中减肥药物α-淀粉酶抑制剂的提纯及其性质研究[D].陕西西安:西北大学,2010.
[25]LI F C,TANG Z H,CUI B W,et al.Hypoglycemic activity of three polysaccharides from Luminaria japonica[J].Chin J Mar Drug,2000,19(5):12-15.(in Chinese)李福川,唐志红,崔博文,等.三种海带多糖的降糖作用[J].中国海洋药物,2000,19(5):12-15.
[26]CHO M L,HAN J H,YOU S G.Inhibitory effects of fucan sulfates on enzymatic hydrolysis of starch[J].LWT-Food Sci Technol,2011,44(4):1164-1171.
[27]KIM K T,RIOUX L E,TURGEON S L.Molecular weight and sulfate content modulate the inhibition ofα-amylase by fucoidan relevant for type 2 diabetes management[J].Pharma Nutrit,2015,3(3):108-114.
[2]ZHOU C S.Extractive technology and physical degradation of polysaccharides from Porphyra yezoensis[D].Zhenjiang:Jiangsu University,2007.(in Chinese)周存山.条斑紫菜多糖提取及物理降解研究[D].江苏镇江:江苏大学,2007.
[3]QIU W F,YANG W J,XUE M.et al.Purification,physicochemical properties and immunomodulatory activity of polysaccharides from Porphyra yezoensis[J].Food Sci,2013,34(23):51-56.(in Chinese)邱伟芬,杨文建,薛梅,等.条斑紫菜多糖的纯化、理化性质及其免疫活性研究[J].食品科学,2013,34(23):51-56.
[4]ZHANG J,YANG X D,WANG W.Study on the anti-tumor effects of Porphyra yezoensis polysaccharide on human hepatocellular carcinoma Bel7402 cells[J].Food Nutr in China,2010(8):82-84.(in Chinese)张杰,杨旭东,王崴.条斑紫菜多糖对人肝癌Bel7402抗肿瘤作用的初步研究[J].中国食物与营养,2010(8):82-84.
[5]HE F,WANG Z H,MA W W,et al.Study on antioxidant activity of polysaccharides with different molecular weights from Pyropia yezoensis in vitro[J].J Shanghai Ocean Univ,2015,24(5):783-788.(in Chinese)何芳,汪之和,马婉婉,等.不同分子量条斑紫菜多糖体外抗氧化活性研究[J].上海海洋大学学报,2015,24(5):783-788.
[6]WANG W M.Study on isolation and purification ofα-amylase inhibitor and its characteristics[D].Tianjin:Tianjin University of Commerce,2010.(in Chinese)王文蒙.α-淀粉酶抑制剂的提取、分离及性质研究[D].天津:天津商业大学,2010.
[7]DILIP C.Phaseolamin compositions and methods for using the same.U.S.Patent 20030059457A1[P].2001-09-25.
[8]LAKSHMANASENTHILS,VINOTHKUMART,GEETHARAMANI D,et al.Fucoidan-anα-amylase inhibitor from Sargassum wightii with relevance to NIDDM[J].Int J Biol Macromol,2015,81(11):644-647.
[9]MITRAKOU A,KELLEY D,MOKAN M,et al.Role of reduced suppression of glucose production and diminished early insulin release in impaired glucose tolerance[J].N Engl J Med,1992,326(1):22-29.
[10]BARON A D.Postprandial hyperglycaemia andα-glucosidase inhibitors[J].Diabetes Res Clin Pract,1998,40(Suppl):S51-S55.
[11]CERIE LLO A.The emerging role of post-prandial hyperglycaemic spikes in the pathogenesis of diabetic complications[J].Diabet Med,1998,15(3):188-193.
[12]DOI T,YOSHIMATSU H,KATSURAGI I,et al.Alphaamylase inhibitor increases plasma 3-hydroxybutyric acid in food-restricted rats[J].Experientia,1995,51(6):585-588.
[13]KIM K T,RIOUX L E,TURGEON S L.Alpha-amylase and alpha-glucosidase inhibition is differentially modulated by fucoidan obtained from Fucus vesiculosus and Ascophyllum nodosum[J].Phytochemistry,2014,98(2):27-33.
[14]PEDROCHE J,YUST M M,GIR仵N-CALLE J,et al.Utilisation of chickpea protein isolates for production of peptides with angiotensin I-converting enzyme(ACE)-inhibitory activity[J].J Sci Food Agr,2002,82(9):960-965.
[15]SUN Y Z,RANG H L,MA R L,et al.Survival of lactic acid bacteria isolated from gut of Epinephelus coioides in mimic gastrointestinal environments[J].J Fishery Sci of China,2010,17(1):128-135.(in Chinese)孙云章,杨红玲,马如龙,等.斜带石斑鱼消化道乳酸菌在模拟胃肠道环境中的存活[J].中国水产科学,2010,17(1):128-135.
[16]RANG Y F.Preparation of white kidney beanα-amylase inhibitor and its application in instant porridge with low GI[D].Wuxi:Jiangnan University,2016.(in Chinese)让一峰.白芸豆α-淀粉酶抑制剂的制备及其在低GI方便粥中的应用[D].江苏无锡:江南大学,2016.
[17]DING G Y,HUANG L G,AN B,et al.Study on LJPS's effect on decreasing hypoglycemic and hypolipidemic[J].Health Med Res Prac,2010,7(4):10-11.(in Chinese)丁国玉,黄连光,安斌,等.海带多糖降血脂、血糖研究[J].保健医学研究与实践,2010,7(4):10-11.
[18]RANG Y F,ZHAO W,YANG R J,et al.Study on activity changes ofα-amylase inhibitor from white kidney beans during processing and digestion[J].Sci Technol of Food Indust,2015,36(17):53-57.(in Chinese)让一峰,赵伟,杨瑞金,等.白芸豆α-淀粉酶抑制剂在加工和消化过程中的活性变化研究[J].食品工业科技,2015,36(17):53-57.
[19]SU Z H.Research on screening marine microorganism of secretingα-amylase inhibitors and purifying the active ingredient[J].Nanning:Guangxi Normal University,2013.(in Chinese)苏照环.海洋生境分泌α-淀粉酶抑制剂微生物的筛选及活性成分纯化的研究[D].广西南宁:广西师范大学,2013.
[20]YU L G.Ultrafiltration,purification and characterization of polysaccharides from Porphyra haitanensis[D].Xiamen:Jimei University,2012.(in Chinese)于立国.坛紫菜多糖超滤纯化与理化性质的研究[D].福建厦门:集美大学,2012.
[21]GONDA R K,TOMODA M,KANARI M,et al.Constituents of the seed of Malva verticillata.III.:Characterization of the major pectic peptidoglycan and oligosaccharides[J].Chem Pharm Bull,1988,36(8):2790-2795.
[22]ZHU X J.Research on extraction,purification and in vitro antitumor activity of polysaccharides from Porphyra yezoensis[D].Nanjing:Nanjing Agricultural College,2008.(in Chinese)朱晓君.条斑紫菜多糖的提取纯化及其体外抗肿瘤活性研究[D].江苏南京:南京农业大学,2008.
[23]LONG S H.The hypoglycemic effect and mechanism of luminaria japonica polysaccharide in type 2 diabetes mellitus model mice[D].Qingdao:Qingdao University,2013.(in Chinese)龙少华.海带多糖在2型糖尿病小鼠中的降糖作用和机制[D].山东青岛:青岛大学,2013.
[24]ZHAO R.Purification and nature research on lose weight medicineα-amylase inhibitor from white kidney beans[D].Xi'an:Northwest University,2010.(in Chinese)赵蓉.白芸豆中减肥药物α-淀粉酶抑制剂的提纯及其性质研究[D].陕西西安:西北大学,2010.
[25]LI F C,TANG Z H,CUI B W,et al.Hypoglycemic activity of three polysaccharides from Luminaria japonica[J].Chin J Mar Drug,2000,19(5):12-15.(in Chinese)李福川,唐志红,崔博文,等.三种海带多糖的降糖作用[J].中国海洋药物,2000,19(5):12-15.
[26]CHO M L,HAN J H,YOU S G.Inhibitory effects of fucan sulfates on enzymatic hydrolysis of starch[J].LWT-Food Sci Technol,2011,44(4):1164-1171.
[27]KIM K T,RIOUX L E,TURGEON S L.Molecular weight and sulfate content modulate the inhibition ofα-amylase by fucoidan relevant for type 2 diabetes management[J].Pharma Nutrit,2015,3(3):108-114.