CN114041607A - 一种食品级大豆蛋白皮克林乳液及其制备方法 - Google Patents
一种食品级大豆蛋白皮克林乳液及其制备方法 Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/185—Vegetable proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
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- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
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- Mycology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- General Preparation And Processing Of Foods (AREA)
Abstract
本发明属于食品工业生物技术领域,涉及一种食品级大豆蛋白皮克林乳液及其制备方法。该皮克林乳液是以脱酰胺化大豆分离蛋白和亚麻籽油为主要成分,纳米级细菌纤维素为稳定剂,水为连续相,使用高剪切均质乳化机均质得到,在室温下放置一个月后乳液仍不分层。本发明制备了以纳米细菌纤维素为稳定剂的功能型乳液,具有良好的稳定性,用于食品领域具有极大的应用潜力。
Description
技术领域
本发明属于食品工业生物技术领域,具体涉及一种食品级大豆蛋白皮克林乳液及其制备方法。
背景技术
亚麻籽油堪称“学生食品”或“知识分子食品”。它的细胞基质中含有木酚素,其含量比其它常用谷类、豆类等要高出100-800倍;亚麻籽中含有人体必须的18种氨基酸,三种维生素(A、E、B1)和8种微量元素。亚麻油中主要成分为α-亚麻酸,其含量高达50-60%,它在人体内可转化为二十碳五烯酸和二十二碳六烯酸,它们为鱼油中的有效活性成分。适量摄入α-亚麻酸可以预防高血压、降血脂、抗肿瘤。但亚麻籽油在应用中易氧化,这可能导致食品变质、异味及营养价值降低等问题。O/W乳液是解决食品工业中油脂稳定性问题的有效方法之一,具有安全性高、制备简单等优点,因此备受关注。
乳液是由两种或多种完全或部分不混溶的液体组成的多相分散系统,其中一种液相(分散相)分布在另一种液相(连续相)中。乳液是动力学不稳定体系,需要依赖乳化剂防止破乳或乳析。与磷脂、吐温等水溶性乳化剂制备的常规乳液不同,皮克林乳液是依靠固体粒子稳定的分散体系。与传统的表面活性剂不同,皮克林乳液中的固体颗粒稳定剂主要是通过不可逆地吸附到油水界面,降低油水界面张力的同时增大空间位阻或提高水相黏度,从而达到稳定乳液的效果。皮克林乳液具有许多优良性质:1.天然生物来源的物质减少或替代了表面活性剂的使用,乳液无毒害,环境友好,符合人们的市场需求。2.具有良好的物理稳定性以及储藏稳定性,受体系环境变化的影响较小。3.通过选用不同种类的固体胶粒或者油相组成,可以调控乳液粒径大小以及流体类型。4.通过选用天然来源的固体颗粒,使得制备的乳液具有良好的环境相容性和生物可降解性。
大豆分离蛋白原料来源广泛、成本较低,并且包含人体必需的20种氨基酸,具有较高的营养价值,是植物蛋白中为数不多的可替代动物蛋白的品种之一。植物蛋白侧链由于富含酰胺基团(谷氨酰胺和天冬酰胺),它们通过氢键稳定蛋白质结构,使得蛋白质之间紧密相连,导致蛋白溶解度很低,影响了植物蛋白乳液的发展。蛋白质谷氨酰胺酶(Protein-Glutaminase,PG,EC:3.5.1.44)是属于谷氨酰胺酶一类的新酶种,只对蛋白质或多肽上的侧链氨基起作用,通过PG处理其侧链上谷氨酰胺键,引起氢键减少和静电斥力的增加,蛋白质空间构象发生变化,在提高植物蛋白饮料特性如溶解性、乳化性、起泡性等方面具有明显作用。并且德国最杰出的的生化学家之一巴德维曾提出过亚麻籽油——蛋白质饮食疗法,此食疗方法使许多身患癌症的病人恢复了健康。亚麻籽油——蛋白质饮食疗法机理是α-亚麻酸与富含含硫氨基酸的蛋白质化合物形成一种脂蛋白化合物,该化合物的双极性可以控制氧的吸收和释放,充足的氧不但能保护正常细胞的功能,同时可以抑制癌细胞的生长和繁殖。
细菌纤维素(Bacterial cellulose,BC)是由木醋杆菌合成的天然纤维素。纤维素等膳食纤维是一种非常重要的食品成分,被称为“第七类营养物质”。细菌纤维素纯度高、结晶度高、吸水能力强、稳定性好、生物安全性突出,目前已被广泛应用于食品、医药、电化学、环境修复等多个领域。
本发明以制备一款亚麻籽油和大豆分离蛋白为主要成分的的功能性食品级乳液为目标,利用细菌纤维素作为功能性食品级乳液的乳化剂,从而制备一种稳定的皮克林乳液。
发明内容
为了解决现有技术中存在的上述问题,本发明的一些实施方式采用以下技术方案:
一种功能性食品级皮克林乳液,由以下重量百分比的组分制成:脱酰胺化大豆分离蛋白5%~15%,细菌纤维素0.05%-0.5%,油脂5%~10%,糖5%~6%,酸味剂0.1%~0.5%,其余为水;其中除油脂的%表示体积百分含量,其余%均为质量百分含量。
作为本发明的一种优选,所述的功能性食品级皮克林乳液,由以下重量百分比的组分制成:脱酰胺化大豆分离蛋白10%~15%,细菌纤维素0.25%-0.5%,油脂8%~10%,糖5%~6%,酸味剂0.1%~0.5%,其余为水。
作为本发明的进一步优选,所述的功能性食品级皮克林乳液,由以下重量百分比的组分制成:脱酰胺化大豆分离蛋白15%,细菌纤维素0.5%,油脂10%,糖5%~6%,酸味剂0.1%~0.5%,其余为水。
作为本发明的一种优选,所述的脱酰胺化大豆分离蛋白通过以下方法制备:
(1)配制5%大豆分离蛋白溶液,加入100U的蛋白质谷氨酰胺酶液,37℃反应1~3h,
(2)反应结束后,将经过蛋白质谷氨酰胺酶改性的大豆分离蛋白溶液冷冻干燥得所述的脱酰胺化大豆分离蛋白。
作为本发明的一种优选,所述的细菌纤维素通过以下方法制备得到:将菌株ATCC53524种子液按6%接种量转接到发酵培养基中,30℃,静置培养5d即可在培养基表面长出膜状物质;将膜状物质从培养基中用镊子取出,冲洗后,在NaOH溶液中煮沸30,滤掉液体;在去离子水中反复浸泡直到滤液呈中性,此时膜状物质呈乳白色,将得到的BC用榨汁机打碎制成悬浮液,将获得的BC超声30min后冷冻干燥得到细菌纤维素。
作为本发明的一种优选,所述的油脂为亚麻籽油。
作为本发明的一种优选,所述的糖选自白砂糖、蔗糖、麦芽糖、果糖;优选白砂糖;所述的酸味剂选自柠檬酸、苹果酸、酒石酸,优选柠檬酸。
作为本发明的一种优选,所述的水为纯化水。
作为本发明的一种优选,所述的功能性食品级皮克林乳液通过以下方法制备的到:脱酰胺大豆分离蛋白溶于纯净水中加入糖、酸味剂、油脂再加入细菌纤维素,使用高剪切乳化机均质后可得到稳定均一的乳液。
本发明所述的功能性食品级皮克林乳液的制备方法,包括脱酰胺大豆分离蛋白溶于纯净水中加入糖、酸味剂、油脂再加入细菌纤维素,使用高剪切乳化机均质后可得到稳定均一的乳液
作为本发明的一种优选,所述乳化机处理参数为180000~20000rpm,3~5min。
作为本发明的一种优选,所述的制备方法还包括将乳液倒入干净的玻璃瓶,135℃杀菌4s,然后迅速冷却到10℃。
与现有技术相比,本发明的有益效果是:
(1)大本发明用蛋白质谷氨酰胺酶处理大豆分离蛋白,获得的脱酰胺化大豆分离蛋白溶解度和乳化性质得到了很大的提高。
(2)亚麻籽油能够弥补人体对不饱和脂肪酸摄入不足的影响,尤其是大豆分离蛋白和亚麻籽油的搭配能够提高身体免疫功能。
(3)本发明采用食品级的细菌纤维素作为乳液稳定剂,细菌纤维素无毒无害,在食品乳液中具有一定的应用前景。
(4)
具体实施方式
以下通过实施例对本发明作进一步的阐述。
实施例1:脱酰胺化大豆分离蛋白制备
称取5.00g大豆分离蛋白,边搅拌边加入适量双蒸水,再加入100U蛋白质谷氨酰胺酶酶液(购自天野酶制剂(上海)有限公司),并用双蒸水定容到100mL。37℃,酶解1~3h后在80℃下加热10min灭酶活。之后用0.10mol/L醋酸溶液透析,最后将样品冷冻干燥。
也可以使用市售蛋白质谷氨酰胺酶替代本实施例蛋白质谷氨酰胺酶,制备脱酰胺化大豆分离蛋白。
实施例2:细菌纤维素(Bacterial Cellulose,BC)的发酵纯化
(1)将菌株ATCC 53524于平板上划线活化4d至长出单菌落;
(2)挑取单菌落于种子培养基中培养24h制备一级种子液;
(3)按8%接种量将一级种子液接种于种子培养基中培养26h制备成二级种子液;
(4)再按6%接种量转接到发酵培养基中,30℃,静置培养5d即可在培养基表面长出膜状物质;
(5)将膜状物质从培养基中用镊子取出,用自来水反复冲洗几次后,在0.1M NaOH溶液中煮沸30min,滤掉液体;
(6)在去离子水中反复浸泡直到滤液呈中性,此时膜状物质呈乳白色,将得到的BC用榨汁机打碎制成悬浮液,将获得的BC超声30min后冷冻干燥得到细菌纤维素。
(7)
实施例3:脱酰胺化大豆分离蛋白功能性质的变化
(1)谷氨酸含量的测定:取120.00g大豆分离蛋白冻干样品置于水解管中,加入15mL 6M盐酸后,充入氮气约15min后立即封瓶,将水解管置于烤箱中110℃烘烤24h,将水解管室温冷却后使用滤纸过滤到25mL容量瓶中,用0.02M盐酸清洗水解管、并将清洗液用滤纸定容至25mL。然后取0.50mL水解液置样品瓶,氮气吹干后加入1.5mL 0.02M盐酸复溶,用0.02mol/L盐酸稀释3倍后,通过0.22um滤膜过滤后用氨基酸自动分析仪测定氨基酸含量。进样量20uL。
(2)溶解度的测定:取0.20g大豆分离蛋白冻干样品溶于20mL磷酸盐缓冲液(pH7.0)中。室温搅拌1h,悬浮液离心后(3000rpm/20min)获得上清液,使用TaKaRa BradfordProtein Assay Kit测定上清液中蛋白质含量。用牛血清白蛋白绘制标准曲线。所有数据都是三次测定的平均值。计算公式如下:
(5)蛋白质样品的乳化活性和乳化稳定性的测定:取0.25g蛋白质样品溶于50mL0.10M PBS缓冲液制备蛋白质分散液。取30mL制备好的蛋白质分散液与10mL大豆油混合,使用均化器15000rpm/mn乳化混合物90s。乳化结束后立即取容器底部20uL乳液到2mL 0.1%SDS溶液中进行稀释,混匀后在500nm下测定吸光度值A0,室温放置15min后再次测定吸光度值A15。以0.1%SDS溶液作为空白。一式三份,计算公式如下:
(6)蛋白质样品的起泡活性和起泡稳定性的测定:取蛋白质样品加入到pH 7.0的0.10M PBS缓冲液中制备蛋白质分散体(0.5%,m/v),然后将15毫升蛋白质样品倒入50毫升量筒中。使用均化器在量筒内以15000rpm/min的速度均质1min。FA计算为混合时体积增加的百分比。FS表示为10分钟后无干扰的剩余泡沫百分比。一式三份,计算公式如下:
表1PG对大豆分离蛋白功能性质的影响
由表1可以看出,经过PG脱酰胺后的大豆分离蛋白功能性质得到显著改善,当蛋白质谷氨酰胺酶添加量为100U/5g蛋白质时,其溶解度、乳化性质和起泡性都得到大幅提高,这是因为大豆分离蛋白经过蛋白质谷氨酰胺酶脱酰胺化后羧基数量增加,由于羧基带负电荷,导致等电点降低,疏水区暴露,其溶解度得到大幅提升;可溶性蛋白具有表面活性,容易吸附油-水界面,从而增加蛋白质的乳化性质;可溶性蛋白更容易在空气泡表面形成刚性膜状结构并且产生更多空气泡,从而起泡性增加。
实施例4:功能性食品级皮克林乳液
按照表2比例制备功能性食品级皮克林乳液,具体步骤如下:脱酰胺化大豆分离蛋白5%~15%,白砂糖5%~6%,柠檬酸0.1%~0.5%,亚麻籽油10%,细菌纤维素0.05%-0.5%,按重量百分比称取各组分,60℃下磁力搅拌均匀后用25MPa的压力均质机均质处理,将饮料倒入干净的玻璃瓶,135℃杀菌4s后迅速冷却到10℃时进行无菌包装得到乳液。
表2功能性食品级皮克林乳液
对比例1
不加脱酰胺大豆分离蛋白,其它组分以及制备方法与实施例3相同。
对比例2
将脱酰胺大豆分离蛋白换成大豆分离蛋白,其它组分以及制备方法与实施例3相同。
对比例3
不加细菌纤维素,其它组分以及制备方法与实施例3相同。
实施例4:测试
取1m L样品溶液置于测量池中,测定温度为25℃,每次测量平衡120s,重复测量3次获得平均值。乳液稳定粒径测试结果如表3。
表3功能性食品级皮克林乳液粒径
乳液的液滴大小影响乳液的稳定性,一般液滴越小乳液稳定性越好。由表3可以看出,本发明实施例2-3的乳液粒径均低于对比例很多,这可能是因为脱酰胺大豆分离蛋白和细菌纤维素可以在O/W界面共存并产生疏水作用与静电相互作用,使乳液中液滴分布更均匀,更稳定。从对比例3不加细菌纤维素与实施例3添加0.5%细菌纤维素中可以看出,细菌纤维素具有很好的乳液稳定效果;从对比例2添加20%的大豆分离蛋白和实施例3添加20%的脱酰胺大豆分离蛋白可以看出大豆分离蛋白因低溶解性和低乳化性使乳液稳定性变差。此结果说明细菌纤维素具有很好的乳化效果,并且经过蛋白质谷氨酰胺酶(PG)改性后的大豆分离蛋白也增加了乳液稳定性。
Claims (10)
1.一种功能性食品级皮克林乳液,其特征在于,由以下百分比的组分制成:脱酰胺化大豆分离蛋白5%~15%,细菌纤维素0.05%-0.5%,油脂5%~10%,糖5%~6%,酸味剂0.1%~0.5%,其余为水;其中除油脂的%表示体积百分含量,其余%均为质量百分含量。
2.根据权利要求1所述的一种功能性食品级皮克林乳液,其特征在于,由以下百分比的组分制成:脱酰胺化大豆分离蛋白10%~15%,细菌纤维素0.25%-0.5%,油脂8%~10%,糖5%~6%,酸味剂0.1%~0.5%,其余为水。
3.根据权利要求2所述的一种功能性食品级皮克林乳液,其特征在于,由以下百分比的组分制成:脱酰胺化大豆分离蛋白15%,细菌纤维素0.5%,油脂10%,糖5%~6%,酸味剂0.1%~0.5%,其余为水。
4.根据权利要求1所述的一种功能性食品级皮克林乳液,其特征在于,所述的脱酰胺化大豆分离蛋白通过以下方法制备:
(1)配制5%大豆分离蛋白溶液,加入100U的蛋白质谷氨酰胺酶酶液,37℃反应1~3h,
(2)反应结束后,将经过蛋白质谷氨酰胺酶改性的大豆分离蛋白溶液冷冻干燥得所述的脱酰胺化大豆分离蛋白。
5.根据权利要求1所述的一种功能性食品级皮克林乳液,其特征在于,所述的细菌纤维素通过以下方法制备得到:将菌株ATCC 53524种子液按6%接种量转接到发酵培养基中,30℃,静置培养5d即可在培养基表面长出膜状物质;将膜状物质从培养基中用镊子取出,冲洗后,在NaOH溶液中煮沸30,滤掉液体;在去离子水中反复浸泡直到滤液呈中性,此时膜状物质呈乳白色,将得到的BC用榨汁机打碎制成悬浮液,将获得的BC超声30min后冷冻干燥得到细菌纤维素。
6.根据权利要求1所述的一种功能性食品级皮克林乳液,其特征在于,油脂为亚麻籽油。
7.根据权利要求1所述的一种功能性食品级皮克林乳液,其特征在于,所述的糖选自白砂糖、蔗糖、麦芽糖、果糖;优选白砂糖;所述的酸味剂选自柠檬酸、苹果酸、酒石酸,优选柠檬酸。
8.根据权利要求1~7中任一项所述的一种功能性食品级皮克林乳液,其特征在于,所述的功能性食品级皮克林乳液通过以下方法制备的到:脱酰胺大豆分离蛋白溶于纯净水中加入糖、酸味剂、油脂再加入细菌纤维素,使用高剪切乳化机均质后可得到稳定均一的乳液。
9.权利要求1~7中任一项所述的功能性食品级皮克林乳液的制备方法,其特征在于,脱酰胺大豆分离蛋白溶于纯净水中加入糖、酸味剂、油脂再加入细菌纤维素,使用高剪切乳化机均质后可得到稳定均一的乳液。
10.根据权力要求9所述的制备方法,其特征在于,所述乳化机处理参数为180000~20000rpm,3~5min。
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