CN116120433A - 一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法及应用 - Google Patents
一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法及应用 Download PDFInfo
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Abstract
本发明涉及生物技术与食品加工领域,具体公开了一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法及应用,该制备方法包括前处理、一次水解与蒸煮、二次水解、酶解、精制和膜过滤,得到的鱼油、鱼粉和鱼胶原蛋白肽的回收率高,且蛋白质含量高,不需要额外再进行除腥和除臭处理,得到的鱼油、鱼粉和鱼胶原蛋白肽无异味,该制备方法能够最大程度地将鱼全身的脂肪、蛋白质和鱼渣分别制成鱼油、鱼胶原蛋白肽和鱼粉,提高了得率和制备效率,水分含量低,易于保存,具有优异的实践价值与广阔的市场前景。
Description
技术领域
本发明属于生物技术与食品加工领域,具体涉及一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法及应用。
背景技术
鯷鱼、沙丁鱼因肌肉组织脆弱,离水后极易受损腐烂,鲜销困难,大都加工晒干,所以常用于提炼鱼油、鱼胶原蛋白肽和制作鱼粉。鱼油是血管的清道夫,对降血脂、降血压有明显的效果和作用。与动物性胶原蛋白相比,鱼类的胶原蛋白分子量是最小的,最有利于人体的吸收。但是,针对一些小型鱼,其具有不易加工、营养价值利用单一,在提取鱼油或鱼胶原蛋白的时候会造成提取资源的浪费。
现有技术中鱼胶原蛋白的制备方法主要包括化学方法和酶法。化学方法是利用酸或碱水解胶原蛋白,虽然方法简单、成本低廉,但水解过程中会破坏L-型氨基酸而形成D-型氨基酸,产物复杂,蛋白质的营养损失大,并且生产过程产生的废水多,生产周期长,不利于工业化生产。与酸法和碱法相比,酶法具有水解效率高、条件温和、易于控制水解进程等优点。专利CN107556364A公开了一种亚临界水辅助酶解提取鲍鱼蛋白肽的方法及产品,技术工艺包括以鲍鱼内脏为原料,利用亚临界水先对其处理萃取蛋白,然后萃取的蛋白再利用果胶酶和蛋白酶进行酶解,通过柱层析分离、纳滤膜浓缩、真空浓缩和喷雾干燥制备鲍鱼蛋白肽粉末,该方法可以大幅度缩短生产时间,降低生产成本。
在鱼油提取工艺中,目前现有技术有以下几种方法:蒸煮法、稀碱水解法和超临界流体萃取法。其中以蒸煮法最为简单,蒸煮法是在蒸煮加热的情况下,使鱼肝的细胞破坏,从而使鱼油分离出来。专利CN108034488A公开了一种深海鱼油的提炼方法,首先通过蒸煮得到鱼脂肪,再通过酶解法将深海鱼脂肪中的鱼油充分的进行提取,接着采用皂化-尿素包合法,鱼油粗提物经皂化除去了少量的饱和脂肪酸,为了进一步的提纯,在皂化后加入了尿素,尿素与饱和脂肪酸和单不饱和脂肪酸形成较稳定的晶体包合物析出,提取物经水洗干燥后即得到含量高达80%的EPA、DHA的深海鱼油。
但是对于同时提高鱼油、鱼粉、鱼胶原蛋白得率的制备方法还鲜有报道。专利申请CN107183571A提供了一种制备粉末鱼油、鱼蛋白肽粉和鱼骨粉的方法,其以鱼类加工下脚料为原料,利用低温酶法进行酶解,分离得到鱼骨微粒、鱼油和鱼蛋白肽液,进一步处理鱼骨微粒以制备鱼骨粉;并向得到的鱼油中添加玉米糊精、小麦水解蛋白,搅拌、浓缩、干燥后制得粉末鱼油;进一步处理鱼蛋白肽液,从而制得鱼蛋白肽粉,该方法使鱼类中蛋白的回收率得到了提高,但是并不能满足市场要求。
发明内容
除非上下文另有明确指示,否则如本文所用的单数形式“一个”、“一种”和“所述”包括单数和复数指示物。通过端点表述的数值范围包括在对应范围内的所有数值和分数,以及所表述的端点。
本发明针对现有技术存在的问题,提供了一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法及应用,该制备方法得到的鱼油、鱼粉和鱼胶原蛋白肽的回收率高,蛋白质的含量高,且不需要额外再进行除腥和除臭处理,得到的鱼油、鱼粉和鱼胶原蛋白肽无异味。
为实现上述目的,本发明采用的技术方案如下:
本发明提供了一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法,其特征在于:
包括以下步骤:
S1、前处理:鱼类破碎后粗磨,再精细研磨为鱼肉膏;
S2、一次水解与蒸煮:向鱼肉膏中添加热水后进行蒸煮,得到鱼浆液,之后进行三相分离,分别得到水相1、沉淀轻相1和沉淀重相1;
S3、二次水解:向得到的沉淀重相1中添加热水进行二次水解后,再次进行三相分离,分别得到水相2、沉淀轻相2和沉淀重相2;将S2得到的水相1和沉淀轻相1分别与水相2和沉淀轻相2混合后,分别得到水相3和沉淀轻相3;
S4、酶解:向得到的水相3中加酶进行酶解处理,调节pH后进行蒸汽灭酶,得到酶解液,进行离心过滤得到水相4和沉淀重相3;将S2的得到的沉淀重相1、S3得到的沉淀重相2和沉淀重相3混合后,得到沉淀重相4;
S5、精制:将S3得到的沉淀轻相3进行物理精制,得到所述的鱼油;将S4得到的沉淀重4相进行气流闪蒸和粉碎,得到所述的鱼粉;
S6、膜过滤:将S4得到的水相中加入质量比为1-3:4的氯化钾和氯化钠后,再依次经过陶瓷膜过滤、超滤膜净化、活性炭纤维素膜过滤、纳滤膜过滤和超滤膜过滤后,进行真空浓缩和喷雾干燥,得到所述的鱼胶原蛋白肽。
优选地,步骤S1所述鱼肉膏的温度为38℃-45℃,鱼肉膏固形物的含量为54%。
进一步优选地,步骤S1所述鱼肉膏的温度为40℃。
优选地,步骤S2所述鱼肉膏与热水的料液比为1.5-3:1;所述热水的温度为92℃-98℃。
进一步优选地,步骤S2所述鱼肉膏与热水的料液比为2.5:1;所述热水的温度为95℃。
优选地,步骤S2所述蒸煮的温度为105℃-115℃,蒸煮的时间为45-55min。
进一步优选地,步骤S2所述蒸煮的温度为110℃,蒸煮的时间为50min。
具体的,步骤S2所述蒸煮的压力为≤0.01MPa。
优选地,步骤S3所述沉淀重相1与热水的料液比为1:2-5;所述热水的温度为85℃-90℃。
进一步优选地,步骤S3所述沉淀重相1与热水的料液比为1:4;所述热水的温度为90℃。
优选地,步骤S4所述的酶为中性蛋白酶、胰蛋白酶、果胶酶、木瓜蛋白酶中的至少一种。
进一步优选地,步骤S4所述的酶为中性蛋白酶和木瓜蛋白酶,所述的中性蛋白酶和木瓜蛋白酶的质量比为2:1-5。
最优选地,步骤S4所述的中性蛋白酶和木瓜蛋白酶的质量比为2:3。
优选地,步骤S4所述酶与水相3的质量比为1:300-500。
进一步优选地,步骤S4所述酶与水相3的质量比为1:400。
优选地,步骤S4所述的pH为6.8-7.5。
进一步优选地,步骤S4所述的pH为7.5。
优选地,步骤S4所述的酶解条件为:温度为50℃-60℃,时间为2-5h。
进一步优选地,步骤S4所述的酶解条件为:温度为55℃,时间为4h。
优选地,步骤S6所述氯化钾和氯化钠的添加量为1-2.5mg/ml水相。
进一步优选地,步骤S6所述氯化钾和氯化钠的添加量为2.2mg/ml水相。
本发明还提供了一种上述的制备方法制备得到的鱼油、鱼粉、鱼胶原蛋白肽在化妆品、保健品、食品、医药中的应用。
相对于现有技术,本发明具有以下有益效果:
本发明提供的制备方法工艺简单、成本低且具有高效益;能够利用全鱼,不浪费资源;通过酶法和膜法的技术结合,使滤渣充分被利用,无废渣产生,对环境污染小;该制备方法能够最大程度地将鱼全身的脂肪、蛋白质和鱼渣分别制成鱼油、鱼胶原蛋白肽和鱼粉,提高了得率和制备效率,且提高了鱼胶原蛋白肽中的蛋白质的纯度,不需要额外再进行除腥和除臭处理,得到的鱼油、鱼粉和鱼胶原蛋白肽无异味,且总体营养价值高,水分含量低,易于保存。
具体实施方式
下面通过具体实施例对本发明进行说明,以使本发明技术方案更易于理解、掌握,但本发明并不局限于此,描述的实施例仅是本申请一部分实施例,而不是全部的实施例。
在本文中所披露的范围的端点和任何值都不限于该精确的范围或值,这些范围或值应当理解为包含接近这些范围或值的值。对于数值范围来说,各个范围的端点值之间、各个范围的端点值和单独的点值之间,以及单独的点值之间可以彼此组合而得到一个或多个新的数值范围,这些数值范围应被视为在本文中具体公开。
基于本申请中的实施例,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它实施例,都应当属于本申请保护的范围。下述实施例中所述实验方法,如无特殊说明,均为常规方法;所述试剂和材料,如无特殊说明,均可从商业途径获得。
实施例1
本发明涉及的一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法,包括以下步骤:
S1、前处理:沙丁鱼(重量180g)破碎后粗磨,再精细研磨为鱼肉膏,鱼肉膏的温度为40℃,鱼肉膏固形物含量为54%;
S2、一次水解与蒸煮:向鱼肉膏中添加95℃的热水(料液比为2.5:1)后进行蒸煮,蒸煮的温度为110℃,蒸煮的时间为50min,蒸煮的压力为≤0.01MPa,之后进行三相分离,分别得到水相1、沉淀轻相1和沉淀重相1;
S3、二次水解:向得到的沉淀重相1中添加90℃的热水(料液比为1:4)进行二次水解后,再次进行三相分离,分别得到水相2、沉淀轻相2和沉淀重相2;将S2得到的水相1和沉淀轻相1分别与水相2和沉淀轻相2混合后,分别得到水相3和沉淀轻相3;
S4、酶解:向得到的水相3中加入中性蛋白酶和木瓜蛋白酶(质量比为2:3)进行酶解处理,其中,酶解条件为:温度为55℃,时间为4h,酶与水相3的质量比为1:400。调节pH为7.0后进行蒸汽灭酶,得到酶解液,进行离心过滤得到水相4和沉淀重相3;将S2的得到的沉淀重相1、S3得到的沉淀重相2和沉淀重相3混合后,得到沉淀重相4;
S5、精制:将S3得到的沉淀轻相3经机械破乳并加热到95℃,得到所述的鱼油;将S4得到的沉淀重4相进行气流闪蒸干燥和粉碎,得到所述的鱼粉;
S6、膜过滤:将S4得到的水相中加入质量比为3:4的氯化钾和氯化钠,氯化钾和氯化钠的添加量为2.2mg/ml水相,再依次经过陶瓷膜过滤、超滤膜净化、活性炭纤维素膜过滤、纳滤膜过滤和超滤膜过滤后,进行真空浓缩和喷雾干燥,得到所述的鱼胶原蛋白肽。
实施例2
本发明涉及的一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法,包括以下步骤:
S1、前处理:沙丁鱼(重量180g)破碎后粗磨,再精细研磨为鱼肉膏,鱼肉膏的温度为38℃,鱼肉膏固形物含量为54%;
S2、一次水解与蒸煮:向鱼肉膏中添加92℃的热水(料液比为1.5:1)后进行蒸煮,蒸煮的温度为115℃,蒸煮的时间为55min,蒸煮的压力为≤0.01MPa,之后进行三相分离,分别得到水相1、沉淀轻相1和沉淀重相1;
S3、二次水解:向得到的沉淀重相1中添加85℃的热水(料液比为1:3)进行二次水解后,再次进行三相分离,分别得到水相2、沉淀轻相2和沉淀重相2;将S2得到的水相1和沉淀轻相1分别与水相2和沉淀轻相2混合后,分别得到水相3和沉淀轻相3;
S4、酶解:向得到的水相3中加入中性蛋白酶和木瓜蛋白酶(质量比为2:1)进行酶解处理,其中,酶解条件为:温度为60℃,时间为3h,酶与水相3的质量比为1:450。调节pH为7.2后进行蒸汽灭酶,得到酶解液,进行离心过滤得到水相4和沉淀重相3;将S2的得到的沉淀重相1、S3得到的沉淀重相2和沉淀重相3混合后,得到沉淀重相4;
S5、精制:将S3得到的沉淀轻相3经机械破乳并加热90℃,得到所述的鱼油;将S4得到的沉淀重4相进行气流闪蒸干燥和粉碎,得到所述的鱼粉;
S6、膜过滤:将S4得到的水相中加入质量比为1:4的氯化钾和氯化钠,氯化钾和氯化钠的添加量为2mg/ml水相,再依次经过陶瓷膜过滤、超滤膜净化、活性炭纤维素膜过滤、纳滤膜过滤和超滤膜过滤后,进行真空浓缩和喷雾干燥,得到所述的鱼胶原蛋白肽。
实施例3
本发明涉及的一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法,包括以下步骤:
S1、前处理:沙丁鱼(重量180g)破碎后粗磨,再精细研磨为鱼肉膏,鱼肉膏的温度为45℃,鱼肉膏固形物含量为54%;
S2、一次水解与蒸煮:向鱼肉膏中添加98℃的热水(料液比为2.5:1)后进行蒸煮,蒸煮的温度为105℃,蒸煮的时间为45min,蒸煮的压力为≤0.01MPa,之后进行三相分离,分别得到水相1、沉淀轻相1和沉淀重相1;
S3、二次水解:向得到的沉淀重相1中添加90℃的热水(料液比为1:5)进行二次水解后,再次进行三相分离,分别得到水相2、沉淀轻相2和沉淀重相2;将S2得到的水相1和沉淀轻相1分别与水相2和沉淀轻相2混合后,分别得到水相3和沉淀轻相3;
S4、酶解:向得到的水相3中加入中性蛋白酶和木瓜蛋白酶(质量比为2:5)进行酶解处理,其中,酶解条件为:温度为50℃,时间为2h,酶与水相3的质量比为1:500。调节pH为6.8后进行蒸汽灭酶,得到酶解液,进行离心过滤得到水相4和沉淀重相3;将S2的得到的沉淀重相1、S3得到的沉淀重相2和沉淀重相3混合后,得到沉淀重相4;
S5、精制:将S3得到的沉淀轻相3经机械破乳并加热90℃,得到所述的鱼油;将S4得到的沉淀重4相进行气流闪蒸干燥和粉碎,得到所述的鱼粉;
S6、膜过滤:将S4得到的水相中加入质量比为2:4的氯化钾和氯化钠,氯化钾和氯化钠的添加量为1.5mg/ml水相,再依次经过陶瓷膜过滤、超滤膜净化、活性炭纤维素膜过滤、纳滤膜过滤和超滤膜过滤后,进行真空浓缩和喷雾干燥,得到所述的鱼胶原蛋白肽。
对比例1
与实施例1相比,区别仅在于:省略步骤S3。其他步骤与实施例1相同。
对比例2
与实施例1相比,区别仅在于:步骤S2中,鱼肉膏与热水的料液比为2.5:3。其他步骤与实施例1相同。
对比例3
与实施例1相比,区别仅在于:步骤S4中,中性蛋白酶和木瓜蛋白酶的质量比为2:7。其他步骤与实施例1相同。
对比例4
与实施例1相比,区别仅在于:步骤S6不同。
S6、膜过滤:将S4得到的水相中加入质量比为3:1的氯化钾和氯化钠后(添加量为2.2mg/ml水相),再依次经过陶瓷膜过滤、超滤膜净化、活性炭纤维素膜过滤、纳滤膜过滤和超滤膜过滤后,进行真空浓缩和喷雾干燥,得到所述的鱼胶原蛋白肽。
其他步骤与实施例1相同。
对比例5
与实施例1相比,区别仅在于步骤S6不同。
S6、膜过滤:将S4得到的水相中加入氯化钠后(添加量为2.2mg/ml水相),再依次经过陶瓷膜过滤、超滤膜净化、活性炭纤维素膜过滤、纳滤膜过滤和超滤膜过滤后,进行真空浓缩和喷雾干燥,得到所述的鱼胶原蛋白肽。
其他步骤与实施例1相同。
效果例1鱼油得率与外观表征
实施例1-3以及对比例1-5的鱼油得率和外观表征情况如表1所示。其中,鱼油得率的计算方法为:鱼油得率%=提取到的鱼油质量/粗脂肪质量×100%,粗脂肪质量由索氏提取法得到。
表1鱼油得率与鱼油外观状态
组别 | 鱼油得率% | 外观表征 | 气味 |
实施例1 | 90.24 | 淡黄色,透明 | 无鱼腥味,无酸味 |
实施例2 | 89.36 | 淡黄色,透明 | 无鱼腥味,无酸味 |
实施例3 | 90.07 | 淡黄色,透明 | 无鱼腥味,无酸味 |
对比例1 | 62.40 | 淡黄色,透明 | 有鱼腥味,稍有酸味 |
对比例2 | 64.33 | 淡黄色,透明 | 有鱼腥味,稍有酸味 |
对比例3 | 89.45 | 淡黄色,透明 | 无鱼腥味,无酸味 |
对比例4 | 89.62 | 淡黄色,透明 | 无鱼腥味,无酸味 |
对比例5 | 89.70 | 淡黄色,透明 | 无鱼腥味,无酸味 |
由表1可知,实施例1-3的鱼油得率明显高于对比例1-2,且外观呈现淡黄色,透明,无鱼腥味无酸味。本发明不需通过酶解工艺但是鱼油得率高,且本发明提供的方法提取得到的鱼油的理化性质达到中国水产行业标准sc/t3502-2016一级粗鱼油的标准。
效果例2鱼胶原蛋白肽的质量
对实施例1-3与对比例1-5制得的鱼胶原蛋白肽粉进行称重,结果如表2所示。
表2各组鱼胶原蛋白肽粉的重量
组别 | 鱼胶原蛋白肽粉重量g |
实施例1 | 16.4 |
实施例2 | 15.2 |
实施例3 | 15.5 |
对比例1 | 7.1 |
对比例2 | 7.4 |
对比例3 | 8.7 |
对比例4 | 7.6 |
对比例5 | 7.2 |
根据表2的数据可知,实施例的鱼胶原蛋白肽粉的重量明显高于对比例,说明本发明特定制备方法制得的鱼胶原蛋白肽粉得率更高。
效果例3蛋白质含量测定
将实施例1-3和对比例1-5制得的鱼胶原蛋白肽粉,分别利用凯氏定氮法检测蛋白质含量,具体结果如表3所示。
表3各组鱼胶原蛋白肽粉中蛋白质含量
组别 | 蛋白质含量% |
实施例1 | 99.8 |
实施例2 | 99.5 |
实施例3 | 99.5 |
对比例1 | 78.1 |
对比例2 | 75.4 |
对比例3 | 65.5 |
对比例4 | 62.3 |
对比例5 | 60.1 |
根据表3的结果可知,本发明实施例1-3中制备方法制备得到的鱼胶原蛋白肽,其蛋白质含量高,通过该方法能够大大提高鱼胶原蛋白肽中的蛋白质含量,纯度高。
效果例4水分含量
利用直接干燥法分别检测实施例1-3和对比例1-5制得的鱼胶原蛋白粉和鱼粉中的含水量,得到表4所示的结果,本发明制得的鱼胶原蛋白肽粉与鱼粉的含水量低,易于保存。
表4鱼胶原蛋白粉和鱼粉中水分含量
组别 | 水分% |
实施例1 | 4.8 |
实施例2 | 4.8 |
实施例3 | 5.0 |
对比例1 | 6.4 |
对比例2 | 6.7 |
对比例3 | 5.2 |
对比例4 | 7.1 |
对比例5 | 7.2 |
最后应当说明的是,以上内容仅用以说明本发明的技术方案,而非对本发明保护范围的限制,本领域的普通技术人员对本发明的技术方案进行的简单修改或者等同替换,均不脱离本发明技术方案的实质和范围。
Claims (12)
1.一种鱼油、鱼粉、鱼胶原蛋白肽的制备方法,其特征在于:包括以下步骤:
S1、前处理:鱼类破碎后粗磨,再精细研磨为鱼肉膏;
S2、一次水解与蒸煮:向鱼肉膏中添加热水后进行蒸煮,得到鱼浆液,之后进行三相分离,分别得到水相1、沉淀轻相1和沉淀重相1;
S3、二次水解:向得到的沉淀重相1中添加热水进行二次水解后,再次进行三相分离,分别得到水相2、沉淀轻相2和沉淀重相2;将S2得到的水相1和沉淀轻相1分别与水相2和沉淀轻相2混合后,分别得到水相3和沉淀轻相3;
S4、酶解:向得到的水相3中加酶进行酶解处理,调节pH后进行蒸汽灭酶,得到酶解液,进行离心过滤得到水相和沉淀重相3;将S2的得到的沉淀重相1、S3得到的沉淀重相2和沉淀重相3混合后,得到沉淀重相4;
S5、精制:将S3得到的沉淀轻相3进行物理精制,得到所述的鱼油;将S4得到的沉淀重4相进行气流闪蒸干燥和粉碎,得到所述的鱼粉;
S6、膜过滤:将S4得到的水相中加入质量比为1-3:4的氯化钾和氯化钠后,再依次经过陶瓷膜过滤、超滤膜净化、活性炭纤维素膜过滤、纳滤膜过滤和超滤膜过滤后,进行真空浓缩和喷雾干燥,得到所述的鱼胶原蛋白肽。
2.根据权利要求1所述的制备方法,其特征在于:步骤S1所述鱼肉膏的温度为38℃-45℃,鱼肉膏固形物含量为54%。
3.根据权利要求1所述的制备方法,其特征在于:步骤S2所述鱼肉膏与热水的料液比为1.5-3:1;所述热水的温度为92℃-98℃。
4.根据权利要求1所述的制备方法,其特征在于:步骤S2所述蒸煮的温度为105℃-115℃,蒸煮的时间为45-55min。
5.根据权利要求1所述的制备方法,其特征在于:步骤S3所述沉淀重相1与热水的料液比为1:2-5;所述热水的温度为85℃-90℃。
6.根据权利要求1所述的制备方法,其特征在于:步骤S4所述的酶为中性蛋白酶、胰蛋白酶、果胶酶、木瓜蛋白酶中的至少一种。
7.根据权利要求6所述的制备方法,其特征在于:步骤S4所述的酶为中性蛋白酶和木瓜白酶,所述的中性蛋白酶和木瓜蛋白酶的质量比为2:1-5。
8.根据权利要求1所述的制备方法,其特征在于:步骤S4所述酶与水相3的质量比为1:300-500。
9.根据权利要求1所述的制备方法,其特征在于:步骤S4所述的pH为6.8-7.5。
10.根据权利要求1所述的制备方法,其特征在于:步骤S4所述的酶解条件为:温度为50℃-60℃,时间为2-5h。
11.根据权利要求1所述的制备方法,其特征在于:步骤S6所述氯化钾和氯化钠的添加量为1-2.5mg/ml水相。
12.权利要求1-11任一项所述的制备方法制备得到的鱼油、鱼粉、鱼胶原蛋白肽在化妆品、保健品、食品、医药中的应用。
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