CN105713943A - 一种用莜麦蛋白粉制备莜麦降糖多肽的方法 - Google Patents
一种用莜麦蛋白粉制备莜麦降糖多肽的方法 Download PDFInfo
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Abstract
本发明提供了一种用莜麦蛋白粉制备莜麦降糖多肽的方法,将脱脂莜麦蛋白粉加入纯净水中,料液比为1:100?500,用1M盐酸调pH值为1.0?3.0,按E/S=1:50?100加入胃蛋白酶,在37℃下酶解1?4h,得到一次酶解液;将一次酶解液,用1M NaoH调pH值为7.0?8.0,然后按E/S=1:50?100分别加入胰蛋白酶、α?糜蛋白酶、弹性蛋白酶,在37℃下酶解1?4h,得到二次酶解液;将二次酶解液,加温至95℃,灭酶活15分钟,喷雾干燥,即得到含有抑制α葡萄糖苷酶水解活性的多肽。本发明工艺简单、生产成本低,产品安全无毒副作用。本发明方法制备的莜麦降糖肽可在制备预防和治疗糖尿病的药物或保健品中应用。
Description
技术领域
本发明涉及莜麦蛋白粉深加工的方法,具体涉及一种用莜麦蛋白粉制备莜麦降糖多肽的方法。
背景技术
莜麦,学名为裸燕麦,为我国特有的粮食作物,也是山西的特色小杂粮,是世界公认的营养价值很高的粮食作物之一,其蛋白含量高出玉米75%,小麦66%,比籼大米、粳大米分别高105.3%和132.8%,莜麦水溶性膳食纤维是小麦的3.5倍,玉米的1.4倍,粳米的7.0倍。莜麦具有多种保健功效,诸如降血脂降胆固醇、抗动脉粥样硬化、控制和降低血糖、降低血压、促进儿童生长延缓老年人衰老的进程,同时预防直肠癌、胆结石和静脉曲张,提高血粘度和血小板聚集。这些功效和莜麦独特的有效成分紧密相关,比如莜麦中亚油酸,芦丁以及人们提到较多的莜麦中可溶性膳食纤维(主要存在于莜麦的麸皮中)。
我国人民在长期食用莜麦的历史中发现,莜麦是一种非常好的药食同源的粮食作物,特别是在作为糖尿病人的食品,起到很好的作用。目前认为莜麦能在糖尿病人食疗起到很好保健作用的是其中的膳食纤维(莜麦粉中可溶性膳食纤维的降糖、降脂作用,中国粮油学报,1998,13(4):37-39)。四氧嘧啶糖尿病小鼠模型试验,揭示与拜糖平一样,莜麦多糖和蛋白均可维持血清中甘油三脂含量处于相对较低水平,莜麦多糖在降糖功效方面优于蛋白,但在改善血清高密度脂蛋白含量上又差于莜麦蛋白与拜糖平。(莜麦中蛋白与多糖成分对四氧嘧啶致糖尿病小鼠血糖、血脂影响比较,食品科技,2013,38(1):66-70)。和蛋白类产品相比,小分子量多肽产品,常常由于兼具多种生理功能比如抗氧化、降糖、降压、抑制癌症、抗菌等,加之相比蛋白吸收快,起效快等特点,成为开发的一个热点。
目前莜麦产品多属初级产品,大多只简单作为一种粮食作物的食品开发,如莜麦饼干,莜麦片等。在莜麦蛋白深度加工利用方面基本处于停滞状态。如何利用莜麦开发功能性食品或药品,提升莜麦综合利用价值,延伸高附加值产业链,在当前具有重要意义。
发明内容
本发明的目的是提供一种用莜麦蛋白粉制备莜麦降糖多肽的方法。
本发明技术方案如下:
一种用莜麦蛋白粉制备莜麦降糖多肽的方法,其特征在于包括以下步骤:
(1)、将脱脂莜麦蛋白粉加入纯净水中,料液比为1:100-500,用1M盐酸调pH值为1.0-3.0,按E/S=1:50-100加入胃蛋白酶,在37℃下酶解1-4h,得到一次酶解液;
(2)、将步骤(1)得到的一次酶解液,用1MNaoH调pH值为7.0-8.0,然后按E/S=1:50-100分别加入胰蛋白酶、α-糜蛋白酶、弹性蛋白酶,在37℃下酶解1-4h,得到二次酶解液;
(3)、将步骤(2)得到的二次酶解液,加温至95℃,灭酶活15分钟,喷雾干燥,即得到含有抑制α葡萄糖苷酶水解活性的多肽。
所述的胃蛋白酶的活力单位为3000-3500U/mg;弹性蛋白酶的活力单位为50U/mg;胰蛋白酶的活力单位为250U/mg;α-糜蛋白酶的活力单位为1200U/mg。
所述步骤(1),步骤(2)中的酶与蛋白底物的比例(E/S)优选为1:50。
本发明方法得到的莜麦降糖多肽分子量低于5000Da的多肽占80%以上,莜麦多肽产品具有抑制α葡萄糖苷酶水解活性,可在制备预防和治疗糖尿病的药物或保健品中应用。
本发明的优点:
1、本发明制备的多肽在体外具有抑制α葡萄糖苷酶水解活性,具有良好抑制多糖代谢的降糖保健功能。
2、本发明结合人体蛋白水解过程,设计酶解工艺,获得特殊功效的多肽产品。
3、本发明制备的多肽产品,分子量控制在5000Da以下占整个产品的80%以上含量。
4、本发明制备技术工艺简单,生产成本低,产品安全无毒副作用,可以用来开发保健品、食品添加剂以及医药中间体。
附图说明
图1为脱脂莜麦蛋白粉酶解前后的Tricine-SDS电泳检测结果,其中:泳道1为蛋白分子量标准;泳道2为一次酶解后多肽产品;泳道3为二次酶解后多肽产品
图2为莜麦脱脂蛋白、莜麦多糖、莜麦水解多肽、阿卡波糖对α-葡萄糖苷酶活性的抑制比较
具体实施方式
下述实施例中所用方法如无特别说明均为常规方法。
实施例1莜麦降糖多肽制备
称取莜麦脱脂蛋白粉与纯净水按重量比1:500配料混合,充分搅拌,用1M盐酸调节pH为1.5,然后以酶与底物(莜麦脱脂蛋白)质量比为1:50加入胃蛋白酶,在37℃下保温1小时,进行酶解反应;然后将所得的酶解液用1MNaOH调节至pH8.0;按酶与底物(莜麦脱脂蛋白)质量比为1:50,分别加入胰蛋白酶、α-糜蛋白酶、弹性蛋白酶,在37℃下二次酶解1h;将二次酶解完毕后的多肽溶液,置于95℃水浴中灭活15min,获得莜麦降糖多肽溶液。
分别取莜麦脱脂蛋白、一次酶解液、二次酶解液样品,进行Tricine-SDS检测,结果如图1。
实施例2莜麦多肽抑制α-葡萄糖苷酶活力的测定
α-葡萄糖苷酶的测活体系:0.5mL反应体系中,底物pNP-α-Glu(4Nitrophenyl-α-D-g1ucopyranosodide)浓度为3×10-4mol/L,还原性谷胱甘肽浓度为8.125×10-5mol/L,酶浓度为0.5U/mL;37℃准确反应10min后,加入终止液(0.1mol/LNa2CO3溶液)2.5mL,测定A410nm。
α-葡萄糖苷酶活性单位定义:在pH6.8,37℃条件下,每分钟释放1μmolPNP为一个酶活单位。
将实施例1制备的莜麦多肽与阿卡波糖(购置于四川绿叶宝光股份有限公司)、莜麦脱脂蛋白(莜麦中蛋白与多糖成分对四氧嘧啶致糖尿病小鼠血糖、血脂影响比较,食品科技,2013,38(1):66-70)、莜麦多糖(购置于宁夏宏德生物技术有限公司)进行对α-葡萄糖苷酶抑制活性测定,结果见图2。莜麦多肽对α-葡萄糖苷酶抑制活性优于阿卡波糖。
Claims (5)
1.一种用莜麦蛋白粉制备莜麦降糖多肽的方法,其特征在于包括以下步骤:
(1)、将脱脂莜麦蛋白粉加入纯净水中,料液比为1:100-500,用1M盐酸调pH值为1.0-3.0,按E/S=1:50-100加入胃蛋白酶,在37℃下酶解1-4h,得到一次酶解液;
(2)、将步骤(1)得到的一次酶解液,用1MNaoH调pH值为7.0-8.0,然后按E/S=1:50-100分别加入胰蛋白酶、α-糜蛋白酶、弹性蛋白酶,在37℃下酶解1-4h,得到二次酶解液;
(3)、将步骤(2)得到的二次酶解液,加温至95℃,灭酶活15分钟,喷雾干燥,即得到含有抑制α葡萄糖苷酶水解活性的多肽。
2.如权利要求1所述的用莜麦蛋白粉制备莜麦降糖多肽的方法,其特征在于,所述的胃蛋白酶的活力单位为3000-3500U/mg;弹性蛋白酶的活力单位为50U/mg;胰蛋白酶的活力单位为250U/mg;α-糜蛋白酶的活力单位为1200U/mg。
3.如权利要求1所述的用莜麦蛋白粉制备莜麦降糖多肽的方法,其特征在于,所述的E/S=1:50。
4.如权利要求1所述方法制备的莜麦降糖多肽在制备预防和治疗糖尿病的药物中应用。
5.如权利要求1所述方法制备的莜麦降糖多肽在制备保健食品中应用。
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Cited By (5)
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CN108949873A (zh) * | 2018-04-19 | 2018-12-07 | 金华市景和科技有限公司 | 从魔芋中提取多肽活性物质的方法 |
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CN115353551B (zh) * | 2022-06-27 | 2024-01-26 | 上海理工大学 | 一种燕麦源促glp-1分泌寡肽及其制备方法和应用 |
CN115521962A (zh) * | 2022-09-26 | 2022-12-27 | 福瑞施生物医药科技(深圳)有限公司 | 水稻多肽组合物及其制备方法和应用 |
CN115521962B (zh) * | 2022-09-26 | 2024-03-15 | 福瑞施生物医药科技(深圳)有限公司 | 水稻多肽组合物及其制备方法和应用 |
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