CN111357865A - 红色食品复合小分子肽及在机体氧化损伤修复中的应用 - Google Patents
红色食品复合小分子肽及在机体氧化损伤修复中的应用 Download PDFInfo
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明提供一种红色食品复合小分子肽及制备方法和在细胞修复中的应用，所述基于红色食品原料的复合小分子肽包括以下原料：赤小豆、红米、红薏米、红南瓜、花生、大枣、山楂、红藜麦、枸杞子、洛神花、红参；所述基于红色食品原料的复合小分子肽的制备方法包括以下步骤：原料粉的制备；发酵产物的制备；酶解产物的制备；复合小分子肽成品的制备。本发明提出的复合小分子肽，具有纯天然，无添加的特点，色泽鲜红诱人，食欲感强，食用人群广，口感好，营养价值高，保质期长，并具有修复受损细胞、延缓衰老、去除氧自由基、补气血及增强免疫力的保健功效。
Description
技术领域
本发明提供一种红色食品复合小分子肽，具有机体氧化修复中的医用用途，同时进一步公开了该小分子肽制备方法，属于涉及药物技术领域。
背景技术
红色食品是指食品为红色、橙红色或棕红色的食品。这类食品富含天然活性植物色素(如番茄红素、花青素、辣椒红素、甜菜红素等)、胡萝卜素、铁、蛋白质和部分氨基酸，是优质蛋白质、碳水化合物、膳食纤维、B族维生素和多种无机盐的重要来源，可以弥补粳米、白面中的营养缺失。科学研究表明，红色食品有治疗缺铁性贫血和缓解疲劳的作用，对乳腺癌等肿瘤疾病有防治作用，给人以兴奋感，有增加食欲，光洁皮肤，增强表皮细胞再生和防止皮肤衰老，预防感冒等作用。经常食用红色食品，可以进一步提高对主食中营养的利用率，山楂、红参等食品还有治疗癌症的功效。被称为“红色生力军。”营养学家认为，红色蔬果最典型的优势在于它们都是富含天然铁质的食物，例如常吃的花生、大枣等都是贫血患者的天然良药，也适合女性经期失血后的滋补。所以，红色蔬果，女人尽可放心多吃。红色食品中还含有致病微生物的“杀手”——巨噬细胞，可以有效地抵御感冒病毒等微生物，增强人体抵抗感冒的能力。
但是上述涉及的红色食品传统的应用方法大多作为普通食品或者提取其中的活性天然植物色素进行开发利用，极少关注其中的蛋白质类成分。科学研究发现，多种食物的蛋白水解物(小分子肽)具有较强的抗氧化能力，而市面上一些普遍的抗氧化剂如维生素E等，虽然能够清除活性氧，但对已造成的损伤修复效果较弱且会产生副作用。本发明对红色食品中的复合小分子肽进行开发利用，弥补了普通抗氧化剂不具有损伤后修复功能的缺陷，更全面地开发利用了红色食品的营养价值。
发明内容
本发明公开一种红色食品复合小分子肽及其制备方法，以赤小豆、红米、红薏米、红南瓜、红衣花生、大枣、山楂、红藜麦、枸杞子、洛神花、红参等众多红色食物为原料，通过发酵及酶解提取的小分子肽，对细胞损伤起到修复作用，解决了传统食用方法对红色食品中蛋白质较少利用的局限性，提高了红色食品的蛋白利用率。
本发明提供了一种红色食品复合小分子肽，其特征在于是由以下原料按以下重量份比制成的：
赤小豆(200～300)；红米(200～300)；红薏米(200～300)；红南瓜(200～300)；花生(100～200)；山楂(50～150)；红糖(50～150)；大枣(50～100)；红藜麦(30～50)；枸杞子(10～30)；猴头菇粉(10～30)；茯苓(10～30)；洛神花(10～30)；红参(10～30)。
本发明所述的一种食品小分子肽的制备方法，包括以下步骤：
1)原料粉的制备：称量上述物料，水清洗干净，烘干去除水分并使用破壁机破壁粉碎，备用；
2)发酵产物的制备：将原料粉倒入发酵桶内并加入面包酵母粉；物料:面包酵母粉体积比＝70～100:1～0.5，密封1小时进行发酵，发酵产物收集备用；
3)酶解产物的制备：发酵液置于混合酶解反应罐，使温度升至53℃，加入木瓜蛋白酶(物料:酶体积比＝70～100:1～0.3)，反应温度45～65℃之间，pH控制在5.0～7.0，反应1h～3h，将木瓜蛋白酶高温灭活处理后的酵素加入风味蛋白酶(物料：酶体积比＝70～100:1～0.3)，pH 6.0左右，温度53℃，反应时间：30～60min，得酶解产物；
4)复合小分子肽成品的制备：将上述酶解后的产物经过胶体磨、均质机处理后得一种食品小分子肽(参见图1：红色小分子肽中肽类成分鉴定分析质谱图)，将该小分子肽进行罐装及杀菌釜杀菌，得成品。
本发明所述的红色食品复合小分子肽是一种活性肽，能有效去除氧自由基，可用于制备细胞修复制品。
【用法用量】口服；2次/日；100ml/次
【适应症】人体细胞修复。
本发明的药理药性如下：
赤小豆：蛋白质含量36％，易于消化，对满足人体对蛋白质的需要具有重要意义；脂肪含量16％，主要含不饱和脂肪酸，吸收率高达95％，除满足人体对脂肪的需要外，还有降低血液中胆固醇的作用；赤小豆含有丰富的维生素、蛋黄素、黑色素及卵磷脂等物质，其中B族维生素和维生素E含量很高，具有营养保健作用，赤小豆中还含有丰富的微量元素，对保持机体功能完整，延缓机体衰老、降低血液粘度、满足大脑对微量物质需求都是必不可少的。
红米：富含的微量元素锌、铜、铁、硒、钼、钙、锰等含量比普通稻(白)米高0.5～3倍；红米还含有黄酮类化合物、生物碱、食品甾醇以及胡萝卜素等药物成份。现代医学证明，红米具有升高血浆高密度脂肪蛋白胆固醇(HD)L和提高机体抗氧化能力的作用；有防治有氧自由基引起的疾病之功效；具有抑制癌细胞、抗肿瘤及免疫复活作用；对老年补钙、儿童增高、长智有益，具有强筋壮骨、补血养颜之功效。这些微量元素和营养物质主要富集在色素中。
红薏米：富含薏仁脂、淀粉、蛋白质以及、维生素等，有丰富的营养，又是普遍、常吃的食物，性味甘淡微寒，有利水消肿、健脾去湿、舒筋除痹、清热排脓等功效，为常用的利水渗湿药；常食可以保持人体皮肤光泽细腻，消除粉刺、斑雀、老年斑、妊娠斑、蝴蝶斑，对脱屑、痤疮、皲裂、皮肤粗糙等都有良好疗效；具有调整免疫抗过敏的功效，是高蛋白质含量之谷类，含有薏仁脂，具有消水肿，帮助消化吸收的作用，而且已被证实具有促进新陈代谢作用，可预防青春痘与皮肤粗糙老化现象的发生，并能减少和淡化色斑。
红参：五加科人参Panaxginseng C.A.Mey.的栽培品，经过蒸制后的干燥根及根茎，于秋季采挖，洗净蒸制后，干燥。红参具有大补元气、复脉固脱、益气摄血的功效，主治体虚欲脱、肢冷脉微、气不摄血、崩漏下血等证。红参在补虚作用方面强于人参，在中医临床上应用很广泛。现代药理学研究显示红参具有抗糖尿病作用，对糖尿病视网膜病变具有保护作用，还具有一定的抗氧化、抗肿瘤、抗衰老等作用。
花生：中含有25％～35％的蛋白，主要有水溶性蛋白和盐溶性蛋白，水溶性蛋白又称为乳清蛋白，占花生蛋白的10％左右，盐溶性蛋白占花生蛋白的90％。花生中的蛋白与动物性蛋白营养差异不大，而且不含胆固醇，花生蛋白的花生蛋白的生物效价为58，蛋白效价为1.7，其营养价值在食品性蛋白质中仅次于大豆蛋白。花生果实还含脂肪、糖类、维生素A、维生素B6、维生素E、维生素K，以及矿物质钙、磷、铁等营养成分，含有8种人体所需的氨基酸及不饱和脂肪酸，含卵磷脂、胆碱、胡萝卜素、粗纤维等物质。花生含有一般杂粮少有的胆碱、卵磷脂，可促进人体的新陈代谢、增强记忆力，可益智、抗衰老、延寿。
大枣：所含有的环磷酸腺苷，是人体细胞能量代谢的必需成分，能够增强肌力、消除疲劳、扩张血管、增加心肌收缩力、改善心肌营养，对防治心血管系统疾病有良好的作用；中医中药理论认为，红枣具有补虚益气、养血安神、健脾和胃等作用，红枣对慢性肝炎、肝硬化、贫血、过敏性紫癜等病症有较好疗效；红枣含有三萜类化合物及环磷酸腺苷，有较强的抗癌、抗过敏作用。大枣能润心肺、止咳、补五脏、治虚损、除肠胃癖气，还能安中养脾、平胃气、通九窍、助十二经等。
红藜麦：富含的维生素、多酚、类黄酮类、皂苷和食品甾醇类物质具有多种健康功效。红藜麦具有高蛋白，其所含脂肪中不饱和脂肪酸占83％，还是一种低果糖低葡萄糖的食物能在糖脂代谢过程中发挥有益功效；红藜麦的营养价值超过任何一种传统的粮食作物，红藜麦是一种全营养完全蛋白碱性食物，红藜麦作为一种藜科食品其蛋白质含量与牛肉相当，其品质也不亚于肉源蛋白与奶源蛋白。藜麦所含氨基酸种类丰富，除了人类必须的9种必须氨基酸，还含有许多非必须氨基酸，特别是富集多数作物没有的赖氨酸，并且含有种类丰富且含量较高的矿物元素，以及多种人体正常代谢所需要的维生素，不含胆固醇与麸质，糖含量、脂肪含量与热量都属于较低水平。
枸杞子：茄科食品枸杞的成熟果实，具有多种保健功效，是卫生部批准的药食两用食物。适量食用有益健康，主要食用功效益精明目，滋补肝肾；适宜人群：虚劳精亏，腰膝酸痛，视力功能下降，眩晕耳鸣。
山楂：具有降血脂、血压、强心、抗心律不齐等作用，同时也是健脾开胃、消食化滞、活血化痰的良药，对胸膈脾满、疝气、血淤、闭经等症有很好的疗效。山楂内的黄酮类化合物牡荆素，是一种抗癌作用较强的药物，其提取物对抑制体内癌细胞生长、增殖和浸润转移均有一定的作用。
洛神花：除了上述几种红色食物，本发明还加入了红南瓜、猴头菇粉、茯苓、洛神花和红糖。猴头菇粉性平、味甘，具有助消化、利五脏的功效。用于治消化不良、胃溃疡、胃窦炎、胃痛、胃胀及神经衰弱等病症；茯苓用于水肿尿少，痰饮眩悸，脾虚食少，便溏泄泻，心神不安，惊悸失眠；洛神花敛肺止咳，降血压，解酒，主肺虚咳嗽，高血压，醉酒。
本发明的积极效果在于：红色食品中通过发酵及酶解得到的食品小分子肽，对过氧化氢导致的A549(人肺癌)细胞损伤起到修复作用；利用小分子肽制成的药物具有临床效果显著，毒副作用小的特点；本发明小分子肽制备方法简单实用；补充营养需求，修复细胞缺损，又可以改善功能状态。通过发酵及酶解分离提取的小分子肽，在细胞修复领域，起着至关重要的作用。
附图说明：
图1为本发明红色小分子肽中肽类成分鉴定分析质谱图。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚，下面将对本发明实施方式作进一步地详细描述。
实施例1
1、称量赤小豆300g、红米300g、红薏米300g、红南瓜300g、花生200g、山楂150g、红糖150g、大枣100g、红藜麦50g、枸杞子30g、猴头菇粉30g、茯苓30g、洛神花30g及红参30g；
2、原料粉的制备：称量上述物料，水清洗干净，烘干去除水分并使用破壁机破壁粉碎，备用；
3、发酵产物的制备：将原料粉倒入发酵桶内并加入面包酵母粉；物料:面包酵母粉体积比＝100:1，密封1小时进行发酵，发酵产物收集备用；
4、酶解产物的制备：发酵液置于混合酶解反应罐，使温度升至53℃，加入木瓜蛋白酶(物料:酶体积比＝100:1)，反应温度45℃之间，pH控制在6.0，反应3h，将木瓜蛋白酶高温灭活处理后的酵素加入风味蛋白酶(物料：酶体积比＝100:1)，pH 6.0，温度53℃，反应时间：60min，得酶解产物；
5、将上述酶解后的产物经过胶体磨、均质机处理后得一种食品小分子肽溶液，将该小分子肽进行罐装及杀菌釜杀菌，得成品。
实施例2
1、称量赤小豆250g、红米250g、红薏米250g、红南瓜250g、花生150g、山楂100g、红糖100g、大枣70g、红藜麦40g、枸杞子20g、猴头菇粉20g、茯苓20g、洛神花20g及红参20g；
2、原料粉的制备：称量上述物料，水清洗干净，烘干去除水分并使用破壁机破壁粉碎，备用；
3、发酵产物的制备：将原料粉倒入发酵桶内并加入面包酵母粉；物料:面包酵母粉体积比＝85:0.8，密封1小时进行发酵，发酵产物收集备用；
4、酶解产物的制备：发酵液置于混合酶解反应罐，使温度升至53℃，加入木瓜蛋白酶(物料:酶体积比＝85:0.6)，反应温度55℃之间，pH控制在7.0，反应2h，将木瓜蛋白酶高温灭活处理后的酵素加入风味蛋白酶(物料：酶体积比＝85:0.6)，pH 6.0，温度53℃，反应时间：45min，得酶解产物；
5、将上述酶解后的产物经过胶体磨、均质机处理后得一种食品小分子肽溶液，将该小分子肽进行罐装及杀菌釜杀菌，得成品。
实施例3
1、称量赤小豆200g、红米200g、红薏米200g、红南瓜200g、花生150g、山楂100g、红糖100g、大枣100g、红藜麦30g、枸杞子10g、猴头菇粉10g、茯苓10g、洛神花10g及红参10g；
2、原料粉的制备：称量上述物料，水清洗干净，烘干去除水分并使用破壁机破壁粉碎，备用；
3、发酵产物的制备：将原料粉倒入发酵桶内并加入面包酵母粉；物料:面包酵母粉体积比＝70:0.5，密封1小时进行发酵，发酵产物收集备用；
4、酶解产物的制备：发酵液置于混合酶解反应罐，使温度升至53℃，加入木瓜蛋白酶(物料:酶体积比＝70:0.3)，反应温度45℃之间，pH控制在5.0，反应1h，将木瓜蛋白酶高温灭活处理后的酵素加入风味蛋白酶(物料：酶体积比＝70:0.3)，pH 6.0，温度53℃，反应时间：30min，得酶解产物；
5、将上述酶解后的产物经过胶体磨、均质机处理后得一种食品小分子肽溶液，将该小分子肽进行罐装及杀菌釜杀菌，得成品。
通过以下实验例证明本发明的小肽对细胞修复作用(本方法参考J EnvironOccup Med 2016,33,233～236)
试验例1
培养A549(人肺癌)细胞株，复苏后培养于改良Eagle(DMEM)高糖培养液中，其中含小牛血清体积分数10％，青霉素(100mg/L)和链霉素(100mg/L)，37℃下置于体积分数为5％的CO2培养箱中无菌培养，消化传代用体积分数为0.25％的胰蛋白酶。
以600μmol/L的H2O2刺激细胞产生损伤。将细胞分为空白对照组，H2O2组(600μmol/L的H2O2损伤24h)。阳性对照组用抗氧化剂N-乙酰半胱氨酸(NAC)处理细胞，即阳性对照前处理组用6mmol/L NAC孵育24h，再加600μmol/L H2O2损伤24h；阳性对照组后处理组用500μmol/L H2O2损伤24h，再加6mmol/L NAC孵育24h。
用25cm2细胞培养瓶培养细胞，药物处理后收集细胞，用4℃磷酸缓冲盐溶液(PBS)洗涤3遍。沉淀用细胞裂解液裂解，4℃1600g离心15min，取上清作为待测样品。用MTT比色法测细胞活力，严格按照试剂盒说明书测定超氧化物歧化酶(SOD)活性、过氧化氢酶(CAT)活性及总抗氧化能力(T-AOC)，使用蛋白浓度试剂盒测定不同组细胞蛋白浓度，校正细胞内SOD、CAT、T-AOC水平。
实验数据均以
进行表示。采用SPSS 19.0软件进行统计学分析，组间均数的比较采用单因素方差分析，两两比较采用Dunnett t检验。检验水准α＝0.05。
小分子肽处理对A549细胞中SOD、CAT和T-AOC的影响(
以蛋白计)
表中数据应使用*等符号标明t检验的结果，如*P<0.05,**P<0.01
实验结果表明，与空白组比较，H2O2组SOD、CAT的活性及T-AOC的能力都下降，差异具有统计学意义(P＜0.05)；与小分子肽组比较，未水解的样品组对SOD活性和T-AOC能力均低于小分子肽样品组；与H2O2组比较，小分子肽处理组对SOD活性和T-AOC能力均达到空白组水平以上；小分子肽处理组修复SOD、CAT活性均比NAC组好。
本发明所述的复合食品小分子肽对过氧化氢致A549细胞损伤起到修复作用；利用小分子肽制成的药物具有临床效果显著，毒副作用小的特点；本发明提取的食物小分子肽制备方法简单实用。
上述本发明实施例序号仅仅为了描述，不代表实施例的优劣。
以上所述仅为本发明的较佳实施例，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (3)
1.一种红色食品复合小分子肽，是由以下原料按以下重量份比制成的：
赤小豆（200～300）；红米（200～300）；红薏米（200～300）；红南瓜（200～300）；花生（100～200）；山楂（50～150）；红糖（50～150）；大枣（50～100）；红藜麦（30～50）；枸杞子（10～30）；猴头菇粉（10～30）；茯苓（10～30）；洛神花（10～30）；红参（10～30）。
2.如权利要求1所述的一种红色食品复合小分子肽的制备方法，其特征在于：
1）原料粉的制备：称量上述物料，水清洗干净，烘干去除水分并使用破壁机破壁粉碎，备用；
2）发酵产物的制备：将原料粉倒入发酵桶内并加入面包酵母粉；物料:面包酵母粉体积比=70～100: 1～0.5，密封1小时进行发酵，发酵产物收集备用；
3）酶解产物的制备：发酵液置于混合酶解反应罐，使温度升至53℃，加入木瓜蛋白酶（物料:酶体积比=70～100: 1～0.3），反应温度45～65℃之间，pH控制在5.0～7.0，反应1h～3h，将木瓜蛋白酶高温灭活处理后的酵素加入风味蛋白酶（物料：酶体积比=70～100: 1～0.3），pH 6.0左右，温度53℃，反应时间：30～60min，得酶解产物；
4）复合小分子肽成品的制备：将上述酶解后的产物经过胶体磨、均质机处理后得一种食品小分子肽、罐装及杀菌釜杀菌，得成品。
3.如权利要求1所述的红色食品复合小分子肽在制备细胞修复制品中的应用。
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| CN114668104A (zh) * | 2022-04-21 | 2022-06-28 | 华戈五谷控股有限公司 | 一种六红谷物片及其制备方法 |
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