CN114847396B - 一种可打印蛋白网络的双凝胶及其制备方法和应用 - Google Patents
一种可打印蛋白网络的双凝胶及其制备方法和应用 Download PDFInfo
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- CN114847396B CN114847396B CN202210433829.5A CN202210433829A CN114847396B CN 114847396 B CN114847396 B CN 114847396B CN 202210433829 A CN202210433829 A CN 202210433829A CN 114847396 B CN114847396 B CN 114847396B
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Abstract
本发明属于食品技术领域，具体涉及一种可打印蛋白网络双凝胶，包括油相和水相；所述油相包含添加油凝胶剂的食用油；所述水相包含添加了植物蛋白和水凝胶剂的盐溶液。本发明提供一种通过乳液模板法制备长期稳定、可3D打印且打印精确度高的双凝胶，将植物蛋白和水凝胶剂加入盐溶液中，形成混合溶液，添加含有油凝胶剂的食用油，通过水浴下高速剪切使其乳化，冷却使其充分凝胶后，所制备得到的水包油双凝胶有较高稳定性，较好的抗形变能力，制备过程简单，容易实现工业化，原料绿色、健康、经济；可用于模拟人造肉中的脂肪组织和结缔组织，避免饱和脂肪酸和反式脂肪酸的同时更好地模拟动物肉的感官和组成。
Description
技术领域
本发明属于食品技术领域，更具体地，涉及一种可打印蛋白网络的双凝胶及其制备方法和应用。
背景技术
随着人类整体社会水平的不断发展，全球肉制品的需求量呈快速增长的趋势。据统计，到 2050 年，全球人口总数将破 100 亿，肉制品的需求量预计将超过 30000 亿美元。为满足人类急速增长的肉制品消费，传统养殖业的产能正面临日益严峻的挑战。同时，随着肉制品大量消耗所带来的环境问题也逐渐加剧。肉制品的消耗大大地增加了对水资源和土地资源的需求，加剧了温室效应。在伦理道德层面上，出于对肉制品的需求，大量动物遭到屠杀，动物的福利和解放等伦理问题引发人们的关注。同时，医学研究证明肉类特别是红肉的过度摄入，极易引发健康问题，如高血糖、高血压、心血管疾病等。近年来，为了解决这些问题，肉类替代物成为人们研究的热点。
现今人造肉的研究主要集中在两个方向：培养肉和植物肉。培养肉被认为是最有前途的肉类替代品，可以直接从干细胞的离体培养中获得，而无需饲养和屠宰动物，这种技术刚刚起步，操作难度较高，生产成本大，难以扩大化生产以及面向市场，并且不同组织分化条件的不同也限制了培养肉的发展。植物肉是以植物蛋白为原料，通过热凝固法、纤维纺丝法和热塑挤压法加工而成的具有与动物肌肉组织相似质构的蛋白制品，常常以大豆蛋白、花生蛋白、小麦蛋白或者豌豆蛋白等植物蛋白为原料。“植物蛋白肉”的生产成本较低，己经有比较成熟的技术，有良好的研究和发展势头。但现有的植物肉在咀嚼过程中口感欠佳，通常会形成一种糊状的质地，因为缺乏动物肉类所具有的结构。
肉类由大约 90% 的肌肉纤维、10% 的脂肪和结缔组织以及不到 1% 的血液组成，可能因肌肉位置和物种而异，植物肉仅仅是通过植物蛋白形成的丝状纤维模拟了肌肉纤维的部分，却不能模拟脂肪组织和结缔组织。更进一步，在真正的肌肉纤维中，肌肉纤维与肌肉纤维间存在的肌外模、肌内膜、肌周膜也是植物肉面临的挑战。以往的研究表明，水凝胶通过合理的结构设计可以增强凝胶的弹性模量、拉伸强度、韧性、硬度和咀嚼性，并具有模拟结缔组织的潜力；在油凝胶体系中，基质强度达到≈1 - 100kpa可满足支撑的机械要求，模仿真实肌肉中结缔组织的结构同时模拟脂肪组织，并且具有热敏性和坚固质地的油凝胶可以在烹饪和咀嚼过程中带来类似动物肉的体验；蛋白网络可以模拟结缔组织中的蛋白组成。
双凝胶通过水凝胶和有机凝胶混合剪切而形成，具有比单凝胶更好的性能，且已经被证明具有剪切稀释行为和触变恢复行为，这是双凝胶在3D打印的性质基础。另一方面，双凝胶在代脂方面的潜力更是被广泛研究。
本方法选用了全植物基的原料。构建出的水包油双凝胶具有优良的打印性质，较高的打印精确度，在植物肉的加工中可模拟动物肉的脂肪组织并打印出大理石纹路，也可与植物肉混合打印模拟动物肉肌肉纤维间的脂肪组织和结缔组织，同时在减少反式脂肪酸、饱和脂肪酸、代脂上都有较好的前景。
授权公开号为CN 108659137 A的中国专利公开了一种具有双凝胶温度的结冷胶及其生产方法和应用，开发了一种低凝胶点的双凝胶且应用于巧克力酱和慕斯中，但制备方法复杂，不利于扩大化生产。
授权公开号为CN 108464948 A的中国专利公开了一种双凝胶体系的高SPF值防晒霜，但添加了各种有机溶剂，主要应用于化妆品领域。
授权公开号为CN 113575923 A的中国专利公开了一种植物肉用热凝胶乳化剂组合物及其用途，使用凝胶乳化剂与肉糜混合提高了植物肉的口感，但没有克服模拟动物肉组织结构这一难题。
Analysis on the printability and rheological characteristics of bigelinks: Potential in 3D food printing文章中探究了不同水油比的双凝胶打印性能，但其可打印范围都在油包水的条件下，本发明利用蛋白网络的乳化特性并在水浴条件下对样品乳化，制备出高内相的水包油双凝胶克服了这一问题。
目前，双凝胶模拟动物肉中脂肪组织和结缔组织的方法尚未有报道，可打印的水包油双凝胶尚未有报道。
发明内容
为了克服上述现有技术的不足，本发明的首要目的是提供一种蛋白网络可打印双凝胶。
本发明的第二个目的是提供上述一种蛋白网络可打印双凝胶的制备方法。
本发明的第三个目的是提供上述一种蛋白网络可打印双凝胶及其在人造肉中的应用。
为了实现本发明的目的，通过以下技术方案进行实现：
一种可打印蛋白网络的双凝胶，包括油相和水相制备成水包油的双凝胶；所述油相包含添加油凝胶剂的食用油；所述水相包含添加了植物蛋白和水凝胶剂的盐溶液；所述植物蛋白可为玉米蛋白、大米蛋白、小麦蛋白、大豆蛋白、米糠蛋白、花生蛋白、豌豆蛋白中的一种或多种；所述水凝胶剂为阿拉伯胶、黄原胶、κ-卡拉胶、果胶、魔芋胶、瓜尔豆胶、海藻酸钠中的一种或多种；所述油凝胶剂可为蜂蜡、米糠蜡、小烛树蜡、单甘脂、葵花蜡、棕榈蜡中的一种或多种。
作为一种优选的技术方案，食用油可为米糠油、菜籽油、大豆油、葵花籽油、花生油、橄榄油、玉米油中的一种或多种。
一种可打印蛋白网络的双凝胶的制备方法，，包括将植物蛋白和水凝胶剂加入盐溶液中，然后添加含有油凝胶剂的食用油，水浴下剪切使其乳化后，冷却24小时使其充分凝胶后，得到双凝胶。
作为一种优选的技术方案，植物蛋白在盐溶液中的浓度为1～10％，水凝胶剂在盐溶液中的浓度为1～10％。
作为一种更优选的技术方案，植物蛋白在盐溶液中的浓度为1～1.5％，水凝胶剂在盐溶液中的浓度为1～1.5％。
作为一种优选的技术方案，食用油浓度为1～90％。
作为一种更优选的技术方案，水油体积比为1~2：3。
作为一种优选的技术方案，食用油中的油凝胶剂浓度为1～30％。
作为一种更优选的技术方案，食用油中的油凝胶剂浓度为1～15％。
作为一种优选的技术方案，水浴温度为65~75℃，剪切乳化条件为9000 r/min的高速均质4 min。
本发明选用了全植物基的原料。构建出的蛋白网络双凝胶具有优良的打印性质，较高的打印精确度，在植物肉的加工中可模拟动物肉的脂肪组织并打印出大理石纹路，也可与植物肉混合打印模拟动物肉肌肉纤维间的脂肪组织和结缔组织，同时在减少反式脂肪酸、饱和脂肪酸、代脂方面许多潜在应用，更重要的是，克服了如今双凝胶在水包油结构下打印性能不好且提高油相后发生相转变的缺点。
与现有技术相比，本发明的有益效果体现在：
本发明采用的原料为全植物基原料，避免了动物肉生产过程中的环境问题和伦理问题；本发明采用植物基蛋白作为骨架模拟动物肉肌肉纤维间的结缔组织，达到进一步模拟动物肉的效果；本发明所制备的蛋白网络双凝胶流变力学性质好，可通过不同种类的油相和较宽的条件范围制备，是安全的、绿色的、可降解的，可食用的，符合大众对健康食品的要求，为代替食品中的饱和脂肪酸、反式脂肪酸提供新选择；且具有较好的可打印性，较高的打印精确度，为3D打印提供了一种新型墨水；本发明所制备的双凝胶利用蛋白的特性形成水包油结构，突破了现有研究中水包油双凝胶打印性差的问题，也解决了油相含量高时双凝胶产生的油包水结构的问题；制备方法简单快速，易于批量生产，为人造肉发展提供新方向。
附图说明
图1是蛋白网络双凝胶制备流程图；
图2是不同水凝胶流变力学测试及触变测试结果图。
图3是不同水凝胶流变力学测试及触变测试结果图。
图4是蛋白网络双凝胶及对照样品外观图，从左到右为乳液、外相结构化的乳液凝胶、内相结构化的乳液凝胶、蛋白网络双凝胶；
图5是实施例1及其对照样品的3D打印效果图，从左到右为乳液、外相结构化的乳液凝胶、内相结构化的乳液凝胶、蛋白网络双凝胶；
图6是实施例1在不同直径喷嘴的3D打印效果图，从左到右为0.6mm、0.8mm、1mm。
图7是实施例7在不同直径喷嘴的3D打印效果图。
具体实施方式
下面将结合本发明实施例，对本发明的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
本发明实施例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，均为可从商业途径得到的试剂和材料。
实施例1
精确称量一定量米糠蛋白、刺槐豆胶和卡拉胶，溶解于pH为5.0的磷酸缓冲溶液中制备：米糠蛋白的质量浓度为1 %，卡拉胶质量浓度、刺槐豆胶质量浓度均为0.75 %的混合溶液，以1:3的水油体积比加入含质量浓度为15 %蜂蜡的菜籽油，然后在70℃水浴下采用剪切乳化机在9000 rmp的剪切速度下剪切4 min使体系乳化，冷却24小时使体系其充分凝胶，得到双凝胶。
如图1所示，图1阐述了本发明的制备流程，首先将蛋白和水凝胶剂混合制备混合溶液，在水浴条件下与含有凝胶剂的植物油混合进行高速剪切（热制备法），由于蛋白质的亲水性，所制备的乳液均为水包油，之后经过24h室温冷却，水相和油相都充分凝胶后得到，蛋白网络双凝胶。
实施例2
精确称量一定量米糠蛋白和魔芋胶，溶解于pH为5.0的磷酸缓冲溶液中制备蛋白浓度、魔芋胶质量浓度都为1.5 %的混合溶液，以1:3的水油体积比加入含质量浓度为10 %蜂蜡的菜籽油，然后80℃水浴下采用剪切乳化机在9000 rmp的剪切速度下剪切4 min使体系乳化，冷却24小时使体系其充分凝胶，得到双凝胶。
实施例3
精确称量一定量花生蛋白和亚麻籽胶，溶解于pH为7.0的磷酸缓冲溶液中制备蛋白浓度、亚麻籽胶质量浓度都为1 %的混合溶液，以1:3的水油体积比加入含质量浓度为4 %单甘脂的花生油，然后70℃水浴下采用剪切乳化机在9000 rmp的剪切速度下剪切4 min使体系乳化，冷却24小时使体系其充分凝胶，得到双凝胶。
实施例4
精确称量一定量米糠蛋白和阿拉伯胶，溶解于pH为7.0的磷酸缓冲溶液中制备蛋白浓度、阿拉伯胶质量浓度都为1 %的混合溶液，以1:3的水油体积比加入含质量浓度为10%小烛树蜡的葵花油，然后90℃水浴下采用剪切乳化机在10000 rmp的剪切速度下剪切5min使体系乳化，冷却24小时使体系其充分凝胶，得到双凝胶。
实施例5
精确称量一定量米糠蛋白、卡拉胶和黄原胶，溶解于pH为7.0的磷酸缓冲溶液中制备蛋白浓度、卡拉胶、黄原胶质量浓度都为1 %的混合溶液，以2:3的水油体积比加入含质量浓度为10 %米糠蜡的菜籽油，然后80℃水浴下采用剪切乳化机在9000 rmp的剪切速度下剪切5 min使体系乳化，冷却24小时使体系其充分凝胶，得到双凝胶。
实施例6
精确称量一定量米糠蛋白和黄原胶，溶解于pH为7.0的磷酸缓冲溶液中制备蛋白浓度、黄原胶质量浓度都为1 %的混合溶液，以1:3的水油体积比加入含质量浓度为5 %葵花蜡的菜籽油，80℃水浴下采用剪切乳化机在9000 rmp的剪切速度下剪切5 min使体系乳化，冷却12小时使体系其充分凝胶，得到双凝胶。
实施例7
精确称量一定量米糠蛋白和刺槐豆胶，溶解于pH为5.0的磷酸缓冲溶液中制备蛋白浓度质量浓度都为1 %、刺槐豆胶质量浓度为1.5 %的混合溶液，以1:3的水油体积比加入含质量浓度为15 %蜂蜡的米糠油，然后80℃水浴下采用剪切乳化机在9000 rmp的剪切速度下剪切4 min使体系乳化，冷却24小时使体系其充分凝胶，得到双凝胶。
测试例1 流变学测试实验方案：
利用HAAKE MARS 40流变仪(Thermo Scientific Inc .,Germany)进行测试，该仪器配置水循环温控系统。
选用20mm几何平板，间隙设置为1mm ,实验温度为20℃，在恒定剪切频率为1Hz，应力为0.1-1000Pa下进行振荡扫描测试，以1-10pa下储能模量的平均值表示凝胶强度（对应图2）。
样品首先经过低剪切步骤 (0.1 s -1 )，然后是高剪切步骤 (10 s -1 )，最后是低剪切步骤 (0.1 s -1 )，每个步骤执行 90 s进行三阶段触变性测试，根据粘度前后变化计算恢复率（对应图3）。
图2和图3筛选了1%浓度下不同水凝胶剂凝胶后的强度、屈服应力和恢复率，在凝胶强度和屈服应力上，由于卡拉胶、卡拉胶+魔芋胶过大的凝胶强度，在挤出后会出现严重的断丝，魔芋胶+黄原胶由于其过大的屈服应力超过3D打印机所提供的挤出力，所以不被选择。
测试例2 3D打印：
挤出式3D打印机(FOODBOT-D1，长兴时印科技有限公司)用于评估不同样品的打印效果。额定挤压应力为71.0 N，打印参数为喷嘴长度2.0 cm、层高0.8 mm、打印速度35 mm/s、喷嘴直径0.8 mm。采用空心六面体作为3D模型，评估打印精度和高度。
图4从左到右为高内相乳液、凝胶乳液（水相凝胶）、凝胶乳液（油相凝胶）、本发明双凝胶体系。
其中，高内相乳液组：1% 浓度米糠蛋白溶液（溶解于pH为5.0的磷酸缓冲溶液中）与菜籽油以水油比1:3比例混合9000rpm剪切4min后冷却24小时使体系其充分凝胶。
凝胶乳液（水相凝胶）：1% 浓度米糠蛋白、0.75%卡拉胶，0.75%刺槐豆胶的混合盐溶液（pH为5.0的磷酸缓冲溶液中）与菜籽油以水油比1:3比例混合9000rpm剪切4min后冷却24小时使体系其充分凝胶。
凝胶乳液（油相凝胶）：1% 浓度米糠蛋白溶液（溶解于pH为5.0的磷酸缓冲溶液中）与含有15%蜂蜡的菜籽油以水油比1:3比例混合9000rpm剪切4min后冷却24小时使体系其充分凝胶。
图5从左到右为高内相乳液、凝胶乳液（水相凝胶）、凝胶乳液（油相凝胶）、双凝胶的打印效果图，可见高内相乳液组由于较低的凝胶强度不能完成自支撑过程导致形状坍塌；凝胶乳液（水相凝胶）由于仅水相凝胶，在挤出后出现明显的漏油现象，虽然具有一定的支撑性，但出现明显的断丝现象（这是水凝胶单独挤出的常见现象）；同样，凝胶乳液（油相凝胶）由于仅油相凝胶，在挤出后出现明显的漏水现象且打印精度不高；对比之下双凝胶在打印后无论是打印精度还是自支撑效果都优于其他组，但仍存在一定漏水现象。
图6是双凝胶在不同直径喷嘴条件下3D的打印效果图，从左到右为0.6mm、0.8mm、1mm，可见在调整喷嘴直径后0.6MM条件下打印的双凝胶精度高，自支撑好，不存在漏水漏油现象。
图7表明实施例7的双凝胶具有极佳的可打印性，且精度高达0.26mm，普遍高于现在乳液体系、乳液凝胶体系、双凝胶体系的打印精度。
综合实施例的实验结果可得，本发明蛋白网络双凝胶与乳液、外相结构化的乳液凝胶、内相结构化的乳液凝胶相比，具有更好的可打印性和打印精度且打印后不会出现漏油现象。
本发明的上述实施例仅仅是为了清楚地说明本发明技术方案的所作的举例，而并非是对本发明的具体实施方式的限定。凡在本发明权利要求书的精神和原则之内所作的任何修改、等同替换和改进等，均应包含在本发明权利要求的保护范围之内。
Claims (5)
1.一种可打印蛋白网络的双凝胶，其特征在于，包括油相和水相制备成水包油的双凝胶；所述制备包括将植物蛋白和水凝胶剂加入盐溶液中，然后添加含有油凝胶剂的食用油，水浴下剪切使其乳化后，冷却24小时使其充分凝胶后，得到双凝胶；所述油相包含添加油凝胶剂的食用油；所述水相包含添加了植物蛋白和水凝胶剂的盐溶液；所述植物蛋白为玉米蛋白、大米蛋白、小麦蛋白、大豆蛋白、米糠蛋白、花生蛋白、豌豆蛋白中的一种或多种；所述水凝胶剂为刺槐豆胶和卡拉胶、卡拉胶和亚麻籽胶、卡拉胶和黄原胶、亚麻籽胶、刺槐豆胶、阿拉伯胶、黄原胶、魔芋胶中的一种；所述油凝胶剂为蜂蜡、米糠蜡、小烛树蜡、单甘脂、葵花蜡、棕榈蜡中的一种或多种；植物蛋白在盐溶液中的浓度为1～10％；水凝胶剂在盐溶液中的浓度为1～10％；食用油浓度为1～90％；食用油中的油凝胶剂浓度为1～30％。
2.根据权利要求1所述可打印蛋白网络的双凝胶，其特征在于，食用油为米糠油、菜籽油、大豆油、葵花籽油、花生油、橄榄油、玉米油中的一种或多种。
3.根据权利要求1所述可打印蛋白网络的双凝胶，其特征在于，水浴温度为65~75℃，剪切乳化条件为9000 r/min的高速均质4 min。
4.权利要求1所述的可打印蛋白网络的双凝胶在制备人造肉中的应用。
5.根据权利要求4所述应用，其特征在于，所述人造肉为植物肉、细胞肉。
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