CN107897942A - 一种基于水溶性低分子量壳聚糖的牡蛎多肽的脱腥方法 - Google Patents
一种基于水溶性低分子量壳聚糖的牡蛎多肽的脱腥方法 Download PDFInfo
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Abstract
本发明涉及一种基于水溶性低分子量壳聚糖的牡蛎多肽的脱腥方法，该方法是在由全牡蛎肉经酶解得到的牡蛎多肽酶解液中，加入一定量的分子量在5000‑20000Da的水溶性低分子量壳聚糖，充分混合，离心取清液即可得到透明无腥味的牡蛎多肽液，再经喷干或冻干得到的牡蛎多肽粉，可迅速溶于水，水溶液清澈透明，为淡黄色液体，无沉淀析出，入口清爽鲜香，无腥味。本发明利用水溶性的壳聚糖，突破了壳聚糖不溶于水的限制，使用范围广泛，且低分子量壳聚糖本身是一种天然无害的低聚糖，还具有免疫调节、防癌、调节肠道菌群、排除体内有毒有害物质等功能活性。
Description
技术领域
本发明属于牡蛎深加工技术领域，具体涉及一种基于水溶性低分子量壳聚糖的牡蛎多肽的脱腥方法。
技术背景
人体对蛋白质的吸收主要以多肽形式进行，分子量在5000Da以下，天然的活性多肽具有各种生理功能和生物活性，如降血压、降血糖、抗菌、抗肿瘤等，活性肽的开发已成为当下研究热点。牡蛎属于药食同源的食品原料，富含各种优质蛋白、锌、硒、牛磺酸、不饱和脂肪酸等大量营养元素，具有“海洋牛奶”的美誉。牡蛎是世界上第一大养殖贝类，但现阶段牡蛎的加工方式多以鲜食、蚝干、罐头、蚝油等初加工为主，精深加工还未得到应有开发，因此，开发具有高附加值的牡蛎功能性食品前景广阔。
腥味使牡蛎及其他水产品的应用受到很大程度的限制，脱腥是水产品加工过程中亟待解决的重点和难点。目前研究较多的脱腥方法主要有：调味剂掩盖、活性炭吸附、酵母发酵、微胶囊包埋、美拉德反应等。调味剂掩盖没有从根本上脱除腥味物质，而且会损失水产品本身的鲜味；活性炭吸附效果虽好，但蛋白损失率很高；酵母发酵过程不易控制，容易发酵过度，且会引入酵母味道；微胶囊包埋的脱腥效果一般，包埋率不好评价；美拉德反应条件要求较高，不易实现。比如公开号为CN 101313736A的专利：“一种淡水鱼片脱腥方法”，是利用乳酸菌两次发酵脱腥，操作繁琐复杂，不易控制；公开号为CN 101230376A的中国发明专利：“一种水产品加工下脚料酶解液脱腥脱色方法”中是利用活性炭吸附脱腥，仅以单一的抗氧化活性回收率为评价指标，没有评价蛋白损失率；文献《美拉德反应用于牡蛎酶解液脱腥的研究_张洁》应用美拉德反应脱腥，反应温度要求100℃，条件要求较高，工业化生产中不易实现。
发明内容
本发明的目的是提供一种牡蛎多肽的脱腥方法，涉及到的牡蛎多肽脱腥方法，脱腥过程操作简单易实现，绿色无污染，蛋白损失率低，小肽产出率高达89％以上，可以从源头上有效脱除腥味物质，并保留了牡蛎的原有风味。
本发明的牡蛎多肽的脱腥方法，是在牡蛎酶解液中添加水溶性低分子量壳聚糖进行脱腥处理；
所述的水溶性低分子量壳聚糖，其制备方法如下：将壳聚糖溶于1％-5％醋酸溶液中，制备1％-5％浓度的壳聚糖溶液，待壳聚糖完全溶解后，缓慢滴入浓度为2％-10％的双氧水溶液，在80℃-90℃下反应4h-10h，反应完成后，用1-4mol/L的氢氧化钠调节其pH值至中性，过滤除去杂质，用3-5倍量的无水乙醇沉淀析出，析出低分子量壳聚糖，冷藏24h后，抽滤，冷冻干燥制成水溶性低分子量壳聚糖；
所述的水溶性低分子量壳聚糖在牡蛎多肽中的添加量为0.5％-5％；
本发明再一个方面提供一种脱腥处理的牡蛎多肽，是在牡蛎酶解液中添加水溶性低分子量壳聚糖进行脱腥处理，离心去除脂溶性的腥味物质和油状漂浮物，得到牡蛎多肽清液，再对清液进行干燥处理获得的；
所述的牡蛎酶解液，其制备方法如下：
1)匀浆：含内脏的全牡蛎肉进行匀浆获得匀浆液；
2)酶解：向牡蛎匀浆液中添加食品级中性蛋白酶或其他蛋白酶(如碱性蛋白酶、木瓜蛋白酶、胃蛋白酶等)，添加量为0.2％-1.5％，酶解温度50-60℃，酶解时间1-4h，酶解完成后加热灭酶获得酶解液；
所述的离心，是在3000-6000r/min离心1-30min；
所述的干燥，是冷冻干燥或喷雾干燥。
本发明使用具有功能活性的水溶性低分子量壳聚糖，突破了壳聚糖不溶于水的限制，使用范围广泛，且低分子壳聚糖本身是一种天然无害的低聚糖，还具有免疫调节、防癌、调节肠道菌群、排除体内有毒有害物质等功能活性，增加了产品的营养性和功能性，另外，利用本发明还可去除大部分牡蛎多肽中的脂肪，进而改善了产品的品质，增加了产品的稳定性。
附图说明
图1：不同pH对低分子壳聚糖溶液透射率的影响图；
图2：低分子量壳聚糖的凝胶渗透色谱图；
图3：脱腥前后牡蛎多肽液挥发性成分的总离子流图
具体实施方式
下面结合实施案例对本发明作进一步详细的描述，但本发明的实施方式不限于此。
实施例1：
水溶性低分子量壳聚糖的制备方法是：5g壳聚糖(分子量为50-300kDa)溶于100mL的2％醋酸溶液中，待壳聚糖完全溶解后，缓慢滴入浓度为5％的双氧水溶液，在80℃下反应6h，反应完成后，用2mol/L的氢氧化钠调节其pH值至中性，过滤除去杂质，用3-5倍量的无水乙醇沉淀析出，析出低分子量壳聚糖，冷藏24h后，抽滤，冷冻干燥。
低分子量壳聚糖的水溶性的估测取决于溶液的浑浊度，0.1g低分子量壳聚糖溶于1％醋酸溶液，配置成浓度为1％的溶液，通过滴加1mol/l的氢氧化钠溶液改变其PH值，在600nm处测定溶液的透射比。如图1所示，本发明的低分子量壳聚糖在pH2-12范围内，溶液的透射率都能达到92％以上，水溶性良好。
采用凝胶渗透色谱(GPC)测定水溶性低分子量壳聚糖的分子量分布，凝胶渗透色谱条件：流动相为0.2mol/LHAc-0.1mol/LNaAc缓冲溶液，微孔过滤器(0.22um)，流速0.8mL/min，柱温30℃，进样量25uL。在不同的凝胶色谱条件下进行GPC分析，结果发现，它们的GPC谱图均具有与图2相似的图形。即出现两个色谱峰，其中3号峰为水峰，2号峰在多种色谱条件下，其保留时间均与2-氨基-2-脱氧-D-葡萄糖盐酸盐相同，是单糖的小分子峰，而1号峰的分子量分布会随壳聚糖的来源、脱乙酰度和降解反应条件的不同而发生变化，本发明制得的水溶性低分子量壳聚糖的分子量分布在5000-20000Da左右。
用此低分子量壳聚糖作为脱腥剂，对牡蛎多肽酶解液进行脱腥处理，具体步骤如下：
(1)匀浆：100g含内脏的全牡蛎肉用组织绞碎机匀浆，不另外添加水，固形物约为15％-20％。
(2)酶解：向牡蛎匀浆中添加食品级中性蛋白酶，添加量1.0％，pH7.0，酶解温度50℃，酶解时间2h，酶解完成后于沸水浴灭酶10min。
(3)去腥：向上述步骤(2)的酶解液中添加浓度为3％的水溶性低分子量壳聚糖，添加量为4％，混合均匀。
(4)离心：将上述步骤(3)的酶解液4800r/min离心1min，得到澄清透明、无腥味的牡蛎多肽清液。
(5)干燥：对上述步骤(4)的牡蛎多肽清液冻干，得到牡蛎多肽粉。
用氯仿甲醇法测定脱腥前后牡蛎多肽酶解液和清液的脂肪含量，清液中脂肪含量仅为6％，脂肪去除率高达94％，用凯氏定氮法测定牡蛎脱腥前后牡蛎多肽酶解液和清液的蛋白含量，清液中多肽回收率为89％，蛋白质的损耗率仅为11％，而活性炭脱腥对蛋白质的损耗率达30％-40％。与活性炭吸附脱腥相比，该发明的优势显而易见，蛋白损耗率低，几乎可以去除全部脂肪，改善了产品品质，引入天然多糖增加了产品的营养性和功能性。
利用气质联用分析脱腥前后的挥发性成分，通过将挥发性化合物的质谱信息与质谱资料库比较，并结合标准品的光谱和保留时间进行定性分析，同时以3-辛醇作为标准物进行定量分析。结果如图3所示，脱腥前后牡蛎酶解液分别检出了44和39种化合物，这些化合物包括碳氢类化合物、呋喃类化合物、醛类化合物、醇类化合物、酮类化合物、酯类化合物和酸类化合物。脱腥前牡蛎多肽液中不愉快气味化合物的质量浓度为32.89ug/mL，脱腥后为4.68ug/mL，不愉快气味的化合物浓度下降了85.77％，表明该发明的脱腥方法能消除或减少绝大部分不愉快气味的化合物，特别是如十七烷、1，3-环辛二烯、2-呋喃甲醇、1，5-辛二烯-3-醇、乙酸、己酸等化合物的含量均有显著减少。
实施例2：
一种基于水溶性低分子量壳聚糖的牡蛎多肽的脱腥方法，其中试规模生产的具体步骤为：
(1)匀浆：100kg含内脏的全牡蛎肉用绞肉机匀浆，不另外添加水，固形物约为15％-20％。
(2)酶解：将上述匀浆投入到200L的酶解罐中，添加食品级中性蛋白酶，添加量1.0％，酶解温度50℃，酶解时间2h，酶解完成后升温至90℃加热灭酶10min。
(3)去腥：向上述步骤(2)的酶解液中添加浓度为3％的水溶性低分子量壳聚糖，添加量为4％，混合均匀。
(4)离心：用平板沉降离心机或卧螺离心机将上述步骤(3)的酶解液离心，得到澄清透明、无腥味的牡蛎多肽清液。
(5)干燥：对上述步骤(4)的牡蛎多肽清液进行喷雾干燥，进风口温度为150-180℃，出风口温度为70-100℃，得到牡蛎多肽粉。
中试生产制得的牡蛎多肽中，脂肪和腥味物质的去除情况及蛋白保留率与实验室结果一致，表明本发明方法具有高的稳定性和可重复性。
Claims (10)
1.一种牡蛎多肽的脱腥方法，其特征在于，所述的方法是在牡蛎多肽液中添加水溶性低分子量壳聚糖进行脱腥处理。
2.如权利要求1所述的方法，其特征在于，所述的水溶性低分子量壳聚糖，其制备方法是将壳聚糖溶于1％-5％醋酸溶液中，制成1％-5％的壳聚糖溶液，待壳聚糖完全溶解后，缓慢滴入双氧水溶液进行反应，反应完成后，用1-4mol/L的氢氧化钠调节其pH值至中性，过滤除去杂质，用3-5倍量的无水乙醇沉淀析出，析出低分子量壳聚糖，抽滤，冷冻干燥制成水溶性低分子量壳聚糖。
3.如权利要求2所述的方法，其特征在于，所述的双氧水溶液的浓度为2％-10％。
4.如权利要求2所述的方法，其特征在于，所述的反应温度为80℃-90℃。
5.如权利要求2所述的方法，其特征在于，所述的反应时间为4-10h。
6.如权利要求1所述的方法，其特征在于，所述的水溶性低分子量壳聚糖在牡蛎多肽中的添加量为0.5％-5％。
7.一种脱腥处理的牡蛎多肽，其特征在于，所述的牡蛎多肽是将牡蛎酶解液由权利要求1所述的方法进行脱腥处理后，离心得到牡蛎多肽清液，再对清液进行干燥处理获得的。
8.如权利要求7所述的牡蛎多肽，其特征在于，所述的牡蛎酶解液，其制备方法如下：
1)匀浆：含内脏的全牡蛎肉进行匀浆获得匀浆液；
2)酶解：向牡蛎匀浆液中添加食品级中性蛋白酶，添加量为0.2％-1.5％，酶解温度50-60℃，酶解时间1-4h，酶解完成后加热灭酶获得酶解液。
9.如权利要求7所述的牡蛎多肽，其特征在于，所述的牡蛎匀浆液中还添加有碱性蛋白酶、木瓜蛋白酶、胃蛋白酶中的任一种或几种。
10.如权利要求7所述的牡蛎多肽，其特征在于，所述的离心，是在3000-6000r/min离心1-30min。
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