CN116235923A - 包含豌豆、米混合物和米糠的高功能蛋白粥预混料及其制造方法 - Google Patents
包含豌豆、米混合物和米糠的高功能蛋白粥预混料及其制造方法 Download PDFInfo
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Abstract
本发明提供了一种粥预混料,其由60重量%至99重量%的米混合物(完整米、米糁、米粉)和1重量%至40重量%的米糠粉组成,其中蛋白质源包括10重量%至50重量%的豌豆粉,包括1%至10%的支链氨基酸,维生素包括75μg RAE/100g的维生素A、10mg/100g的维生素C和0.15mg/100g的维生素B2(核黄素),并且矿物质包括80mg/100g的钙、10mg/100g的锌和60mg/100g的镁。本发明还提供了一种用于制造高功能蛋白粥预混料的方法。本发明可以提供具有增加的蛋白质含量的功能蛋白粥,以使老年患者能够快速康复,并获得柔软的吞咽特性和咀嚼感。
Description
相关申请的交叉引用
本申请要求2021年12月8日提交的韩国专利申请号10-2021-0174435的优先权,其整体通过引用并入本文。
发明背景
1.技术领域
本发明涉及用于制造包含豌豆、米混合物和米糠的高功能蛋白粥预混料的方法。更具体地,本发明涉及:关于包含60重量%至99重量%(wt.%)的由完整米、米糁和米粉组成的米混合物以及1重量%至40重量%的稳定米糠粉的粥预混料,一种高功能蛋白粥预混料,其包含10重量%至50重量%的豌豆粉作为蛋白质源,1%至10%的支链氨基酸,70μg/100g至80μg/100g的维生素A、0.14mg/100g至016mg/100g的维生素B2(核黄素)、8mg/100g至12mg/100g的维生素C、70mg/100g至90mg/100g的钙、8mg/100g至12mg/100g的锌和50mg/100g至70mg/100g的镁;及其制造方法。
2.背景技术
米粥是通过将包含淀粉的谷物(作物)或食品材料粉碎,然后用水煮以使颗粒流淌或变软而制备的具有易于消化的特征的食物。因此,粥不仅是提供给患者或婴儿的膳食,而且经常被正常人以保健食品或喜爱的食品如鲍鱼米粥、鸡肉粥、蔬菜粥、红豆粥等形式食用。随着医疗技术的不断发展和国民健康保险福利的广泛传播,平均寿命延长使得老年人口急剧增加,从而进入超老龄化社会。大多数老年患者依靠老年医院或老年疗养院、日夜护理中心等,并享受国家健康保险公司的长期护理支持基金的福利。这种老年护理中心目前在全国范围内超过5,000个,并且趋于迅速增加。现在,在老年住院患者中,每天三餐食用粥的患者接近30wt.%左右。传统的制备粥的方法需要时间将米浸泡在水中,然后煮,并且需要烹饪人员,因此导致向老年护理中心的租户大量供应粥的困难。此外,该粥作为代餐缺乏营养,并且具有导致吞咽不便的问题(即喉咙处不柔软)。替代地,有一种粥制造方法,其将米精细粉碎,然后煮。然而,在这种情况下,粥的生产就像米布丁一样,因此吃粥的人可能很少咀嚼,因而降低了咀嚼的感觉,从而导致作为代餐的利用率降低。因此,现在实际上需要开发一种功能粥,其可以在短时间内制备并提供柔软的质地和咀嚼感,同时可以用作具有用于老年患者的营养满意度和免疫力的代餐或用作婴儿食品。
动物和植物有两种类型的免疫系统,即先天免疫和获得性免疫。其中,先天免疫是一种细胞或免疫机制,用于通过非特异性免疫方法(自然免疫、先天免疫)保护宿主免受感染。这涉及免疫系统立即应答,而没有关于特定病原体或抗原的记忆。体内包含的吞噬细胞如巨噬细胞或抗生素在体内免疫中起重要作用,其中从先天免疫细胞分泌的代表性细胞因子包括白细胞介素6(IL-6)、白细胞介素12(IL-12)和肿瘤坏死因子α(TNF-α)。另一方面,获得性免疫作为特异性免疫(适应性免疫、获得性免疫)在暴露于抗原后被诱导,此后,当将来暴露于相同的抗原时,记忆的免疫应答被诱导。在更高生命形式的情况下,在暴露于病原体的个体的感染早期阶段获得性免疫不起作用的时期,先天免疫优先作用,随后表现出更有效的获得性免疫。对于获得性免疫过程,B细胞、T细胞等起主导作用。特别地,从获得性免疫细胞分泌的代表性细胞因子包括白细胞介素2(IL-2)和干扰素-γ(IFF-γ)。
豌豆是一种双子叶植物,特别是玫瑰树豆中的一年生/两年生作物。豌豆通常用作与米一起煮的豆类,就像红豆或芸豆一样,用于年糕和/或糖果的豆粉。此外,未成熟和小的青豌豆用于罐头食品,嫩豆荚用作蔬菜,并且其叶子和茎用于牲畜饲料。豌豆包含蛋白质、复合碳水化合物、纤维、矿物质、维生素和抗氧化物质,因此被用作营养物质的极好来源。豌豆包含一般成分,例如8.1g水,20.7g蛋白质,65.1g碳水化合物,1.3g脂质和2.8g灰分。特别地,豌豆中包含的膳食纤维(每100g豌豆中6g)有助于身体废物排泄和肠道健康;包含欧米茄3脂肪酸而促进脑细胞的作用;包含脂肪酸例如亚油酸和油酸而降低胆固醇水平,同时协助钠排泄,从而有助于血液血管健康;异黄酮成分作为抗氧化剂,从而消除有害氧(通常是自由基);包含皂苷和维生素K,并且有助于抑制癌细胞的生长。豌豆包含14%至31%的粗蛋白、15%至25%的白蛋白、49%至70%的球蛋白、11%的谷蛋白、5%的醇溶谷蛋白、8.9%的赖氨酸、11%的谷氨酰胺、半胱氨酸、精氨酸等。此外,豌豆包含矿物质和维生素B1而防止肌肉流失,并且富含氨基酸而有助于蛋白质合成,从而有助于增加肌肉。众所周知,豌豆有益于肠道健康,大脑健康,血管健康,增加肌肉,预防癌症等。
大豆由35%至47%的蛋白质、30%的碳水化合物和15%至20%的脂肪组成,并且是富含膳食纤维、磷、铁、钙和维生素B1的营养食品。大豆种类繁多,具有高含量的对生命保护很重要的必需氨基酸,并且可以提供高质量的蛋白质。另外,大豆降低葡萄糖在胃和肠道中的吸收率以抑制糖尿病的发生,富含作为植物雌激素的异黄酮而有助于预防骨质疏松症。此外,大豆富含维生素E,有助于血液循环和防止衰老,并且可激活双歧杆菌促进排便,从而在便秘治愈中有效。据报道,大豆中包含的一些功能性成分,如染料木苷(genistin)、皂苷、植酸、植物甾醇等,有助于减少乳腺癌、直肠癌、结肠癌、肺癌、胃癌、前列腺癌等。此外,大豆中的卵磷脂可有效增强大脑的注意力,改善痴呆和健忘症,防止大脑衰老。大豆中的异黄酮可以降低体内胆固醇,抑制作为雌性激素的雌激素的过度分泌,从而在缓解更年期症状的同时预防乳腺癌和卵巢癌。大豆对膳食以及预防成人疾病和脱发(秃头症)是有效的,特别地,黑豆壳中包含的花青素色素含有强抗氧化成分,以消除有害的活性氧,从而有助于抗癌作用和衰老预防。特别地,黑豆已被选为世界10种食物之一,并且被称为能量食品。
支链氨基酸(BCAA)是在侧链上具有支链碱的氨基酸,例如缬氨酸、亮氨酸、异亮氨酸等,也可以是疏水性氨基酸。这有助于活性中心(例如酶)中结合位点的形成。亮氨酸促进α螺旋结构的形成,而缬氨酸和异亮氨酸通常存在于β结构中。就人类营养而言,这些属于必需氨基酸。三种类型的BCAA不通过肝脏,而是直接进入肌肉进行新陈代谢,并为肌肉提供必要的能量,从而帮助肌肉的形成和协助肌肉细胞的活动。事实上,老年患者或手术患者摄入蛋白质降解为氨基酸的速率较低,这反过来又降低了将氨基酸重建为肌肉蛋白质的速率。特别地,支链氨基酸中的亮氨酸被用作治疗肌肉减少症的辅助剂。
作为与豌豆相关的传统艺术之一,韩国专利注册公开号10-2028652(标题为“/> (包含豌豆粉、发酵米糠、米胚芽、韩国当归根提取粉、人参提取粉、蜂蜜粉和可可粉的粉状多营养补充食品)”)涉及粉状形式的多营养补充食品,其包含豌豆粉、发酵米糠、米胚芽、韩国当归根提取粉、人参提取粉、蜂蜜粉和可可粉。此外,韩国专利早期公开公开号10-2021-0008845(标题为“ (基于水解豌豆蛋白的营养组合物)”)提供了一种营养组合物,其包含至少一种蛋白质源,包括占总组合物的2.25重量%-6.5重量%的豌豆蛋白水解产物;以及至少一种植物化学提取物、至少一种脂肪酸、至少一种有机成分和至少一种益生元纤维。此外,韩国专利早期公开公开号10-2021-0020080(标题为“/> (基于完整豌豆蛋白的营养组合物)”)涉及一种用于肠道和口服施用的基于完整豌豆蛋白的营养组合物,其由有机和植物基组分形成。该营养组合物包含豌豆蛋白、植物化学提取物、脂肪酸、不含8种超级过敏原的有机成分、和益生元纤维。该营养组合物以液体形式提供,以便被施用于肠道和口服。然而,如上所述的这种传统艺术在技术配置方面与本发明主要不同。
现有技术文献
专利文献
(专利文献1)韩国专利注册公开号10-2028652
(专利文献2)韩国专利早期公开公开号10-2021-0020080
(专利文献3)韩国专利早期公开公开号10-2021-0008845
非专利文献
(非专利文献1)TEXTURIZATION OF PULSE PROTEINS:PEAS,LENTILS,AND FABABEANS by TAEHOON KIM(论文)。2018。提交给德克萨斯A&M大学研究生和专业研究办公室,部分满足哲学博士学位的要求。第17页。
发明内容
本发明人提出了韩国专利注册公开号10-1441565,其涉及包含米混合物和稳定米糠的功能粥预混料及其制造方法。该专利涉及一种粥预混料,其包含60重量%至99重量%的由完整米、米糁和米粉组成的米混合物,以及1重量%至40重量%的稳定米糠粉。然而,该米混合物中所含的蛋白质仅在7g/100g至8g/100g之间,这对用于膳食的粥来说是低蛋白质含量,并且这不足以作为代餐。因此,作为为了加强粥预混料中蛋白质的缺乏的重复研究的结果,本发明通过采用在各种具有高粗蛋白含量的豆类中具有理想物理性质和良好口感的豌豆并添加它,此外,还添加支链氨基酸、维生素和矿物质到预混料中,完成了一种用于制造功能粥预混料的方法。
[表1]
干豌豆中的蛋白质和氨基酸(%)
划分 | 粗蛋白 | 白蛋白 | 球蛋白 | 谷蛋白 | 醇溶谷蛋白 |
豌豆 | 14-31 | 15-25 | 45-70 | 11 | 5 |
扁豆 | 23-31 | 17 | 51 | 11 | 4 |
蚕豆 | 36 | 2 | 74 | 18 | 4 |
本发明人公开的韩国专利注册公开号10-1441565(标题为“ (包含米混合物和稳定米糠的功能粥预混料及其制造方法)”)包括:(S1)将米粉碎,再将其分为完整米、米糁和米粉,其分别具有三(3)种颗粒尺寸;(S2)将完整米、米糁和米粉按2.5-3.5:4.5-5.5:1.5-2.5的重量比混合;(S3)将60重量%至99重量%的混合的米混合物与1重量%至40重量%的稳定米糠粉混合,从而生产粥预混料。为了加强作为上述韩国专利注册公开号10-1441565的“/> (包含米混合物和稳定米糠的功能粥预混料)”的缺点的蛋白质缺乏,本发明人从动物蛋白或豆类(大豆)中的鸡蛋蛋白或乳清蛋白中选择豌豆,在植物蛋白中选择豌豆和扁豆。豌豆以糊状或豆粉的形式广泛使用,因此消费者很熟悉。此外,除了良好的营养、物理性质和味道外,豌豆是对米混合物具有理想亲和力的物质。特别是对于老年患者,考虑了消化率较低,咀嚼力不足和吞咽不适。虽然豆类具有高脂肪含量的优点,但应去除脂肪,以便将豆类用作预混料的物质。此外,在脱脂大豆的情况下,涉及硬组织、咀嚼性和吞咽性差。因此,比豆类更软并且味道更好的豌豆被用作预混料的物质。对豌豆进行漂烫和快速冷却以抑制酶促反应,从而防止营养成分的损失,同时保留其风味。此外,在漂烫和快速冷却后,将经处理的豌豆粉碎并酶分解,以便于老年患者吸收。因此,本发明提供了一种用于制造高功能蛋白粥预混料的方法,其包括:将豌豆在纯化水中洗涤和浸泡,去除水,在高温下对豌豆进行灭菌,漂烫,在低温下快速冷却经漂烫的豌豆,并将快速冷却的豌豆粉碎的过程;添加豌豆粉10倍的纯化水以使所述粉均质化,并使用已知和市售的蛋白水解酶水解粉末的过程;加热水解产物以终止酶促反应,然后离心以得到豌豆酶水解产物的过程;干燥酶水解产物,筛选并去除杂质的过程;以及混合豌豆粉、米混合物(完整米、米糁、米粉)、米糠粉、支链氨基酸和微量维生素和矿物质从而产生粥预混料的过程。
对于上述植物原料,可以对选自具有高蛋白质含量的豆类(大豆)、扁豆和蚕豆中的任意一种或两种或更多种进行预处理,例如脱脂、粉碎等,然后,可以部分替代豌豆,即可以与豌豆组合使用。替代地,分离的大豆蛋白或植物蛋白可以与豌豆组合使用。
本发明的粥预混料可用于提供高功能蛋白粥,其由米混合物、米糠、豌豆蛋白和支链氨基酸组成,可用维生素和矿物质增强。特别地,豌豆的灭菌、漂烫然后快速冷却的过程可以抑制酶促反应,以防止营养成分的损失,同时有助于保持风味。此外,使用蛋白水解酶进行水解可以增加蛋白质含量,使得免疫能力和体重得以维持,特别地,由于柔软的吞咽特性和咀嚼感,预混料可以容易地被老年患者吸收并允许老年患者快速恢复。因此,本发明可用作老年人保健食品、成人代餐、婴儿营养食品等。
附图说明
本发明的上述和其他目的、特征和其他优点将从结合附图的以下详细描述中更清楚地理解,其中:
图1是本发明的工艺流程图;
图2示出了粥预混料中一氧化氮(NO)的生成量,其中N为阴性对照,RP(1)表示(米混合物+米糠),而RP(2)表示(米混合物+米糠+豌豆+支链氨基酸+维生素+矿物质);
图3示出了关于先天免疫的RP(1)和RP(2)的IL-6和IL-12、TNF-α中每一种的表达水平;
图4示出了关于获得性免疫的RP(1)和RP(2)的IL-2、TNF-γ中每一种的表达水平;
图5A示出了使用高温杀菌和漂烫技术制备的豌豆粉,图5B示出了包含豌豆粉的粥预混料,图5C示出了使用包含豌豆粉的粥预混料烹制的粥;以及
图6A是通过扫描电子显微镜(SEM)x1000放大率(S-3400N,日本日立)拍摄的米粉的照片,示出了硬质微粒的凝结,并且图6B是豌豆粉的照片,示出了软的和圆形颗粒单独分离的现象。
具体实施方式
通过引入豌豆漂烫技术制造功能性蛋白质粥预混料的方法如下所述。具体地,上述制造方法可包括:在将豌豆在纯化水中洗涤和浸泡后,去除豌豆中的水;在高温下灭菌的同时将豌豆漂烫,在低温下快速冷却经漂烫的豌豆,并将豌豆粉碎;将纯化水添加到粉碎的豌豆粉中以使其均质化,然后,向其添加市售的蛋白水解酶,以便在振荡恒温水浴(“水浴”)中水解豌豆粉;将水解产物在高温下加热使酶失活,然后离心,从而得到豌豆酶水解产物;干燥豌豆酶水解产物,筛选至期望尺寸,并去除杂质;将豌豆粉与米混合物(完整米、米糁、米粉)、米糠粉、支链氨基酸、维生素和矿物质混合,从而产生粥预混料。在这方面,豌豆的漂烫可以抑制豆类或蔬菜的酶促反应,以防止营养成分的损失,同时保持风味。此外,当豌豆在漂烫后进行酶分解,快速冷却和压碎豌豆时,豌豆可能具有改善的质地以增强物理性质和味道,从而容易被老年患者吸收。此外,酶处理的豌豆在烹饪后表现出优异的营养,同时具有相对较高的硬度,从而改善消化率和咀嚼感。
1)豌豆的洗涤、浸泡和脱水
在市场上购买的豌豆(韩国首尔的Garak市场)用自来水或纯化水彻底洗涤。将洗涤的豌豆浸泡在18℃至25℃的纯化水中3小时至4小时以充分浸软,然后将豌豆放在柳条托盘上除水4小时至5小时。
2)豌豆的漂烫、快速冷却和粉碎
在将豌豆在78℃至82℃下漂烫和灭菌20分钟至30分钟后,将灭菌的豌豆在0℃至5℃下快速冷却1小时至2小时。将快速冷却的豌豆粉碎到90目至100目。在这方面,豌豆的漂烫可以抑制豆类或蔬菜的酶促反应,以防止营养成分的损失,同时保持风味。此外,在漂烫和快速冷却后获得的豌豆可以被赋予柔软的质地。
3)豌豆粉的酶分解
在将豌豆粉用水以1:9-10的比例均质化后,可以将市售蛋白水解酶以0.1%(w/v)至1.0%(w/v)的量添加到上述混合物中,然后pH 4至5下在40℃至50℃的振荡恒温水浴中水解10分钟至30分钟。本文使用的市售蛋白水解酶可以是选自碱性蛋白酶(Alcalase)、复合蛋白酶(Protamex)、中性蛋白酶(Neutrase)、益瑞蛋白酶(Esperase)、风味蛋白酶(Flavourzyme)、菠萝蛋白酶(Bromelain)、Laminex Super 3G或Maxazyme(NovoNordiskBiochem Biochem)的蛋白水解酶。优选地,以0.2%(w/v)至0.4%(w/v)的量添加选自菠萝蛋白酶1200GDU、Laminex Super 3G、复合蛋白酶或风味蛋白酶的任意一种或两种或更多种以进行酶分解,然后在95℃至100℃下加热30秒至50秒以使酶失活。终止水解产物的酶促反应后,在11,000x g至12,000x g下离心14至16分钟,从而获得豌豆酶水解产物。
[表2]
通过豌豆的酶分解控制硬度的实例
可以看出,当在漂烫和快速冷却豌豆之后进行酶分解时,硬度随着酶浓度的升高和活化时间的增加而降低。
4)酶水解产物的干燥和筛选,以及杂质的去除
在将豌豆酶水解产物在50℃至60℃下干燥7小时至8小时以达到8%至9%的含水量之后,将水解产物筛选至0.4mm至0.6mm或更小,同时去除杂质,例如废金属或不锈钢、石材、塑料等。
5)混合以制备粥预混料
将10重量%至50重量%的蛋白质含量为40重量%至60重量%的豌豆粉、60重量%至99重量%的米混合物(完整米、米糁、米粉)、1重量%至40重量%的米糠粉、1%至10%的支链氨基酸以及微量的维生素和矿物质混合以产生粥预混料。本文中,支链氨基酸是混合比例为1:0.5:0.5的亮氨酸、异亮氨酸和缬氨酸的混合物,并且可以以1%至10%的量被包含。此外,所述维生素可包括维生素A、B、C、D、E和K等。优选地,包含维生素A、B和C中的任意一种或多种。特别地,基于100重量份的粥预混料,粥预混料可包含70μg RAE/100g至80μgRAE/100g的维生素A、8mg/100g至12mg/100g的维生素C和0.14mg/100g至0.16mg/100g的维生素B2(核黄素)。上述矿物质可包含硒、锌(或氧化锌)、镁、铜等,但它们并不局限于此。优选地,包含钙、锌和镁中的任意一种或多种。特别地,基于100重量份的粥预混料,可以通过添加70mg/100g至90mg/100g的钙,8mg/100g至12mg/100mg的锌和50mg/100g至70mg/100g的镁的混合物来产生粥预混料。矿物质还可以帮助免疫和抗炎功能。这些维生素和矿物质可以满足韩国食品和药物管理局推荐的日常需求的10%或更多。
[表3]
质量比较
<实施例1-4>
将1kg豌豆在室温下用纯化水洗涤,并以1:3的比例浸泡在纯化水中,然后在柳条托盘上去除水并对豌豆脱水。将豌豆在80℃下漂烫30分钟以使酶灭活。此外,将经漂烫的豌豆在0℃下快速冷却1小时。将漂烫和快速冷却的豌豆在55℃下干燥8小时,以达到8%的含水量,然后将其粉碎到90目至100目。将水和豌豆粉以10:1的比例混合并使其均质化后,将选自菠萝蛋白酶1200GDU、Laminex Super 3G、复合蛋白酶或风味蛋白酶的任意一种或两种或更多种以0.1%(w/v)至1.0%(w/v)的量添加以在pH 4至5下在40℃至50℃的振荡恒温水浴(“水浴”)中进行水解10分钟至30分钟,然后在95℃至100℃下加热30秒至50秒以使酶失活。终止水解产物的酶促反应后,在11,000×g至12,000×g下离心14分钟至16分钟,从而得到豌豆酶水解产物。随后,将10wt%至50wt%的粉状豌豆粉与60wt%至99wt%的米混合物,1wt%至40wt%的稳定米糠粉,1%至10%的支链氨基酸,3种维生素,特别地,75ugRAE/100g或更多的维生素A、10mg/100g或更多的维生素C、0.15mg/100g或更多的维生素B2(核黄素),和三种类型的矿物质,特别地,80mg/100g或更多的钙、10mg/100g或更多的锌和60mg/100g或更多的镁(6种成分的混合物)混合,从而产生粥预混料。关于本发明所采用的混合比例,为了补充老年人的消化、咀嚼和营养功能,上述三种维生素和三种矿物质可以分别以满足推荐的日常需要的量加入。此外,关于本发明的“高功能蛋白粥预混料(牛肉和蔬菜粥)”,其中将米混合物、米糠、豌豆蛋白和BCAA氨基酸混合,本发明人曾向韩国功能性食品研究所(2021年11月15日)要求分析蛋白质和支链氨基酸成分,并且获得结果证明在粥预混合剂中包含18.08%的粗蛋白,3028mg/100g亮氨酸,1494.93mg/100g异亮氨酸和1665.30mg/100g缬氨酸。可以看出,这样的蛋白质和氨基酸是非常有利的营养素,以满足推荐日常需求的约30%。此外,本发明人曾向水原女子大学(2017年6月13日)要求分析上述“牛肉和蔬菜粥预混料”的营养成分,并且获得证明7.08mg/100g的蛋白质含量的结果。与上述结果相比,可以看出使用豌豆和支链氨基酸的本发明具有非常高的蛋白质含量,提高至现有粥预混料的3.9倍,从而证明上述组分是优良的营养成分。
本发明的粥预混料是提供给运动或活动较少的患者的食物,因此,不应造成免疫系统的损害,以便通过饮食供应保持健康状态,同时不影响体重的变化。因此,进行了免疫测试(体外)和体重变化(体内)的实验。
<实验例1>免疫测试(体外)
对于包含米混合物和米糠的粥预混料(1)和包含与米混合物和米糠混合的豌豆、支链氨基酸、维生素和矿物质的高功能粥预混料(2),进行实验以研究免疫系统和体重、白细胞的变化,以及是否损伤DNA。
为了分析通过本发明的粥预混料(1)和高功能粥预混料(2)改善先天免疫的效果,分别测量了根据上述提取物的施用的一氧化氮(NO)生成量,以及与先天免疫相关的每种细胞因子的表达水平,特别是白细胞介素IL-6和IL-12以及肿瘤坏死因子α(TNF-α)。
1)测量一氧化氮生成量
将小鼠RAW 264.7细胞以1x105个细胞/孔的浓度放入48孔板中后,将LPS用作阳性对照,并且每个样品以100μg/mL的量处理。孵育24小时后,获得细胞培养上清液,并使用Griess试剂测量NO生成量。分别向每只小鼠施用RP(1)(米混合物+米糠)和RP(2)(米混合物+米糠+豌豆+维生素+矿物质),然后测量小鼠细胞中的一氧化氮生成量。测量结果如图2所示,表明RP(2)中的NO生成量比RP(1)高60%或更高。
2)先天免疫相关细胞因子的测量
如下进行IL-6表达水平的测量。将小鼠RAW 264.7细胞以1x105个细胞/孔的浓度放入48孔板中后,将LPS用作阳性对照,并且每个样品以100μg/mL的量处理。48小时孵育后,获得细胞培养上清液,并使用ELISA试剂盒测量IL-6生成量。
如下进行IL-12表达水平的测量。将小鼠RAW 264.7细胞以1x105个细胞/孔的浓度放入48孔板中后,将LPS用作阳性对照,并且每个样品以100μg/mL的量处理。48小时孵育后,获得细胞培养上清液,并使用ELISA试剂盒测量IL-12生成量。
如下进行TNF-α表达水平的测量。将小鼠RAW 264.7细胞以1x105个细胞/孔的浓度放入48孔板中后,将LPS用作阳性对照,并且每个样品以100μg/mL的量处理。48小时孵育后,获得细胞培养上清液,并使用ELISA试剂盒测量TNF-α生成量。
分别向每只小鼠施用RP(1)(米混合物+米糠)和RP(2)(米混合物+米糠+豌豆+维生素+矿物质),然后分别测量小鼠细胞中每种先天免疫细胞因子的生成量,即IL-6,IL-12和TNF-α。测量结果如图3所示。证实本发明的粥预混料仅表现出IL-6和IL-12细胞因子表达的增加,而TNF-α的表达略有增加。
3)获得性免疫相关细胞因子的测量
为了分析通过本发明的粥预混料(1)(米混合物+米糠)和高功能粥预混料(2)(米混合物+米糠+豌豆+维生素+矿物质)改善获得性免疫的效果,测量了根据上述提取物的施用的细胞因子IL-2和干扰素-γ(IFN-γ)的表达水平。
如下进行IL-2表达水平的测量。将小鼠脾细胞以2x106个细胞/孔的浓度放入48孔板中后,将ConA用作阳性对照,并且每个样品以100μg/mL的量处理。48小时孵育后,获得细胞培养上清液,并使用ELISA试剂盒测量IL-2生成量。
如下进IFN-γ表达水平的测量。将小鼠脾细胞以2x106个细胞/孔的浓度放入48孔板中后,将ConA用作阳性对照,并且每个样品以100μg/mL的量处理。48小时孵育后,获得细胞培养上清液,并使用ELISA试剂盒测量IFN-γ生成量。关于分别施用RP(1)(米混合物+米糠)和RP(2)(米混合物+米糠+豌豆+维生素+矿物质)的小鼠细胞,获得性免疫细胞因子的生成量的测量结果如图4所示。其证明,在施用根茎(Rhizoma)联合植物提取物的小鼠细胞中,获得性免疫细胞因子IL-2和IFN-γ的表达水平增加。从上述结果可以看出,本发明的粥预混料表现出一氧化氮生成量的增加,以及与先天免疫和获得性免疫相关的所有细胞因子的表达水平的增加。因此,本发明的粥预混料适用于老年患者食品。
<实验例2>体重变化(体内)
使用本发明的粥预混料(1)和高功能蛋白粥预混料(2),使用实验动物(小鼠)进行体重变化的实验。
分别使用本发明的粥预混料RP(1)(米混合物+米糠)和高功能蛋白粥预混料RP(米混合物+米糠+豌豆+支链氨基酸+维生素+矿物质)替代约20%的一般饲料(LabDiet,实验室啮齿动物日粮5001),从而制备试验饲料,同时使用制备的一般饲料作为对照。由6周龄并且体重为20g至25g的10只雄性C57BL/6小鼠摄取每种饲料2周。1周和2周后,分别测量摄取RP(1)、RP(2)和对照的小鼠的体重变化。此外,2周后,对所有小鼠进行尸检和血液收集,然后使用血细胞分析仪测量白细胞的数量。此外,通过彗星测定法评估脾脏外周免疫细胞的DNA损伤程度。由于每个个体的日平均饲料摄入量为4.0g/天,所以14天的摄入量为56.0g,并且所有实验组(1)和(2)以及对照表现出显著差异。
[表4]
体重变化
划分 | 饲料(1) | 饲料(2) | 一般饲料(对照) |
1周后 | 无变化 | 无变化 | 无变化 |
2周后 | 1%增加 | 3%增加 | 无变化 |
1)体重变化
与作为对照的摄入一般饲料的小鼠相比,摄入20%的替代饲料(1)和(2)的小鼠体重明显增加。从上述结果可以证实,与摄入包含粥预混料(1)的饲料的实验组以及摄入一般饲料(对照)的实验组相比,摄入包含高功能蛋白粥预混料(2)的饲料的实验组的体重增加了1%至3%。因此,可以看出本发明提供了一种优良的高功能蛋白粥预混料,其增加了实验小鼠的体重。
2)白细胞的数量
实验期2周后,进行尸检并测量白细胞数量。作为测量的结果,两个实验组和对照的白细胞数量都在正常范围内(1.8-10.7K/μL)。因此,关于白细胞的数量,实验组(1)和(2)与对照之间没有显著差异。
3)DNA损伤
作为进行彗星测定以确定DNA损伤程度的结果,证实两个实验组和对照在体内DNA损伤方面均未显示出任何变化。
从上述实验结果,通过细胞系实验(体外)和动物实验(体内)可以看出,本发明的粥预混料可以赋予免疫能力并增加体重,从而可以提供优良的粥预混料。
本发明可以提供一种高功能蛋白粥,其包含米混合物、米糠、豌豆蛋白和支链氨基酸,并用维生素和矿物质增强。特别地,在漂烫、快速冷却和粉碎豌豆后,使用蛋白水解酶对经处理的豌豆进行水解以增加蛋白质含量,使得产生的粥容易被老年患者吸收,从而允许快速恢复,同时保持免疫力和体重。特别地,本发明的粥可用作保健食品、代餐和营养餐。因此,本发明在工业上是适用的。
Claims (10)
1.一种高功能蛋白粥预混料,其包含:60重量%至99重量%的由完整米、米糁和米粉组成的米混合物,和1重量%至40重量%的稳定米糠粉,其中所述高功能蛋白粥预混料还包含豌豆粉、支链氨基酸、维生素和矿物质。
2.根据权利要求1所述的预混料,其中,除了60重量%至99重量%的由完整米、米糁和米粉组成的米混合物和1重量%至40重量%的稳定米糠粉之外,所述高功能蛋白粥预混料还包含10重量%至50重量%的豌豆粉作为蛋白质源,
其中所述支链氨基酸包括1%至10%的混合比例为1:0.5:0.5的亮氨酸、异亮氨酸和缬氨酸的混合物,
所述维生素包括70μg/100g至80μg/100g的维生素A、0.14mg/100g至0.16mg/100g的维生素B2(核黄素)和8mg/100g至12mg/100g的维生素C,以及
所述矿物质包括70mg/100g至90mg/100g的钙、8mg/100g至12mg/100g的锌和50mg/100g至70mg/100g的镁。
3.根据权利要求1所述的预混料,其中所述豌豆通过将豌豆在78℃至82℃下漂烫和灭菌20分钟至30分钟,将其在0℃至5℃下快速冷却1小时至2小时,并对经处理的豌豆进行粉碎和均质化,然后使用蛋白水解酶进行水解并将水解产物干燥以80目至100目的颗粒尺寸获得。
4.根据权利要求1所述的预混料,其中所述预混料包含豌豆、所述米混合物和米糠的混合物,并且其中所述蛋白质源包括与所述豌豆粉组合的选自分离的大豆蛋白、脱脂大豆粉、脱脂扁豆粉或脱脂蚕豆粉中的任意一种或多种。
5.一种用于制造高功能蛋白粥预混料的方法,包括:
将豌豆在纯化水中洗涤和浸泡,去除水,在高温下对所述豌豆进行灭菌,对其进行漂烫,在低温下快速冷却经漂烫的豌豆,并将快速冷却的豌豆粉碎;
向豌豆粉中添加纯化水以使所述粉均质化,并使用市售的蛋白水解酶水解所述粉;
加热水解产物使所述酶失活,然后离心得到豌豆酶水解产物;
将所述酶水解产物干燥,筛选,并且去除杂质;以及
将豌豆粉、米混合物(完整米、米糁、米粉)、米糠粉、支链氨基酸和微量维生素和矿物质混合,从而制备粥预混料。
6.根据权利要求6所述的方法,其中所述豌豆通过将豌豆在纯化水中洗涤和浸泡,去除水,将所述豌豆在78℃至82℃下漂烫和灭菌20分钟至30分钟,将其在0℃至5℃下快速冷却1小时至2小时,然后将经处理的豌豆粉碎到90目至100目而获得。
7.根据权利要求5所述的方法,其中将所述豌豆粉通过添加10倍的纯化水进行均质化,并使用已知和市售的蛋白水解酶进行水解以产生水解产物,并且
将所述水解产物加热以终止酶促反应,然后以11,000x g至12,000xg离心14分钟至16分钟,从而得到豌豆酶水解产物。
8.根据权利要求5所述的方法,其中将所述豌豆酶水解产物在50℃至60℃下干燥7小时至8小时以达到8%至9%的含水量,并且筛选至90目至100目,然后去除杂质。
9.根据权利要求5所述的方法,其中所述粥预混料由60重量%至99重量%的米混合物(完整米、米糁、米粉)和1重量%至40重量%的米糠粉组成,并且还包含10重量%至50重量%的蛋白质含量为40重量%至60重量%的豌豆粉
其中所述支链氨基酸包括1%至10%的混合比例为1:0.5:0.5的亮氨酸、异亮氨酸和缬氨酸的混合物;所述维生素包括70μg/100g至80μg/100g的维生素A、0.14mg/100g至016mg/100g的维生素B2(核黄素)和8mg/100g至12mg/100g的维生素C;并且所述矿物质包括70mg/100g至90mg/100g的钙、8mg/100g至12mg/100g的锌和50mg/100g至70mg/100g的镁,以及
通过将上述成分混合来制备粥预混料。
10.一种高功能蛋白粥预混料,其通过根据权利要求5至9任一项所述的方法制造。
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