CN109602896A - 一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法和应用 - Google Patents
一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法和应用 Download PDFInfo
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- CN109602896A CN109602896A CN201910036842.5A CN201910036842A CN109602896A CN 109602896 A CN109602896 A CN 109602896A CN 201910036842 A CN201910036842 A CN 201910036842A CN 109602896 A CN109602896 A CN 109602896A
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Abstract
本发明涉及食品和药品技术领域,公开了一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法和应用。本发明所述组合物由胶原蛋白肽和弹性蛋白肽组成;所述胶原蛋白肽由胶原蛋白原料经胃蛋白酶或胰蛋白酶酶解制成,所述弹性蛋白肽由弹性蛋白原料经木瓜蛋白酶和/或复合蛋白酶Protamex酶解制成。本发明通过特定的工艺制备获得了特定分子量区间的弹性蛋白肽和胶原蛋白肽,两者在合适的比例下组成的组合物,能够以较少的使用量同时显著提高受损皮肤中弹性蛋白和胶原蛋白的含量,并显著提高透明质酸,羟辅氨酸含量和降低MMP3含量,同时还能够抑制皮肤炎症因子。
Description
技术领域
本发明涉及食品和药品技术领域,特别涉及一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法和应用。
背景技术
弹性蛋白是一种构成弹性纤维的蛋白质,其广泛分布在生物体内的各个组织和脏器中,主要存在于颈韧带、血管、肺、皮肤等有弹性的组织中。而这类组织不仅需要弹性,同时拉伸强度也是必不可少的,而且弹性蛋白对形态的构成和维持发挥重要的作用。弹性蛋白富含甘胺酸、丙氨酸、缬氨酸等疏水性氨基酸,而且在弹性蛋白生物合成过程中,其前驱物质弹性蛋白原通过赖氨酰氧化酶形成了氨基酸、锁链素、异锁链素,并对由四个赖氨酸残基构成的交联结构进行干预,所以弹性蛋白对保持组织结构的稳定性和弹性有所帮助。
随着年龄增长,肌肤中的弹性蛋白的成分减少,连带地降低对维持紧致肌肤所需要的网络支持,结果就是肌肤变得松弛没,轮廓变得模糊。过了25岁,弹性蛋白就停止生产,皮肤中的弹性蛋白也就不断的流逝,皮肤开始松弛、有细纹等老化现象。科学研究证实,皮肤的年龄由弹性蛋白的百分比决定,只有弹性蛋白能够提供和保持皮肤的二维弹性。
胶原蛋白和弹性蛋白是在皮肤真皮层里位于表皮下方的两种蛋白质。生产这两种物质的特殊细胞结缔组织——成纤维细胞。胶原蛋白和弹性蛋白形成的“基础的皮肤”是胶原蛋白“支持”的表皮,胶原蛋白能增加皮肤弹性,提拉紧致,提供活性。这两种蛋白质能保持水分、滋润肌肤、保湿,使皮肤光滑,防止皱纹的出现。
不少人认为,留住肌肤青春的保养重点就是如何补充胶原蛋白,其实弹性蛋白更为重要。弹性蛋白在肌肤中扮演橡皮筋的角色,让肌肤有伸展和褶合的能力,其功用就如床垫中的弹簧,负责维持与支撑肌肤的弹性。因此,弹性蛋白在维持肌肤弹性方面担当着重要的角色。弹性蛋白决定着皮肤的弹性和柔软性,对光线等理化因素引起的皮肤生理性老化过程,具有防止老化和促进再生的作用。护肤品中如果同时拥有弹性蛋白和胶原蛋白成分的话,对保持肌肤年轻、细腻、弹性会有非常好的作用。
很早以前弹性蛋白就作为美容目的使用,不过是将牛的颈韧带为原料作为化妆品素材使用。近年来,随着BSE和家禽传染病等的发现,其安全性遭到了质疑,所以亟待寻求可代替的原料。
在这样的背景之下,人们探讨将牛韧带、牛心管、鲣鱼动脉球、猪心脏大动脉等作为原料制造弹性蛋白肽的方法,通过口服和护肤来实现美容的目的。然而目前受弹性蛋白肽的工艺制备以及倍受青睐的因素影响,其成本较高,而能够达到增加弹性蛋白含量目的的前提是需要摄入足够量的弹性蛋白肽,这无疑会增加消费者的成本。此外,除了年龄带来的皮肤松弛等问题,外在环境导致的皮肤炎症也是影响衰老的一大因素,如何能够使弹性蛋白高效的发挥其作用并抑制皮肤炎症是一个有广阔经济价值的研究课题。
发明内容
有鉴于此,本发明的目的在于提供一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法,使得所述组合物能够具备协同作用,同时显著增加弹性蛋白含量以及胶原蛋白含量;
本发明的另外一个目的在于提供一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法,使得所述组合物能够同时显著提高透明质酸,羟辅氨酸含量和降低MMP3含量;
本发明的另外一个目的在于提供一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法,使得所述组合物能够显著抑制皮肤炎症因子,从而实现抑制皮肤炎症的目的;
本发明的另外一个目的在于提供上述组合物在制备食品和/或药品中的相关应用。
为实现上述发明目的,本发明提供如下技术方案:
一种胶原蛋白肽和弹性蛋白肽的组合物,由胶原蛋白肽和弹性蛋白肽组成;所述胶原蛋白肽由胶原蛋白原料经胃蛋白酶或胰蛋白酶酶解制成,所述弹性蛋白肽由弹性蛋白原料经木瓜蛋白酶和/或复合蛋白酶Protamex酶解制成。
作为优选,所述胶原蛋白肽和弹性蛋白肽的质量比为(1.6-3.2):(0.075-0.2);更优选为(1.6-2.4):0.1;在本发明具体实施方式中,所述胶原蛋白肽和弹性蛋白肽的质量比为1.6:0.1或2.4:0.1,例如3.2g胶原蛋白肽和0.2g弹性蛋白肽,或者4.8g胶原蛋白肽和0.2g弹性蛋白肽。
在本发明具体实施方式中,所述胶原蛋白原料选自鳕鱼,所述弹性蛋白原料可选自牛韧带、牛心管、鲣鱼动脉球和猪心脏大动脉等弹性蛋白含量较高的原料中的一种或两种以上。
同时,本发明还提供了所述组合物的制备方法,包括:
胶原蛋白原料加胃蛋白酶或胰蛋白酶酶解,酶解液过1~2μm滤膜,滤液喷雾干燥获得胶原蛋白肽;
弹性蛋白肽原料去除表面脂肪和杂质,加木瓜蛋白酶和/或复合蛋白酶Protamex酶酶解,酶解液过0.45~1μm滤膜,滤液喷粉获得弹性蛋白肽;
混合胶原蛋白肽和弹性蛋白肽获得所述组合物。
进一步地,所述制备方法包括:
胶原蛋白原料干燥粉碎,加水以及1%胃蛋白酶或胰蛋白酶酶解,酶解液过1~2μm滤膜,滤液喷雾干燥获得胶原蛋白肽;
弹性蛋白肽原料加NaOH溶液浸泡去除表面脂肪和杂质,清洗并绞碎,加水并调节pH7.5~8.5,加入木瓜蛋白酶和/或复合蛋白酶Protamex酶解,酶的加入量为0.4~0.8%,酶解液采用板框过滤,并过0.45~1μm滤膜,滤液浓缩后喷粉,获得弹性蛋白肽;
混合胶原蛋白肽和弹性蛋白肽获得所述组合物。
在本发明具体实施方式中,所述制备方法包括:
取胶原蛋白肽原料剪碎,置于80℃烘箱烘干12小时,后用粉碎机粉碎成粉末,加入20-30倍量水,溶液升温至60℃±5℃,向溶液中加入1%胃蛋白酶/胰蛋白酶,酶解4~8小时,升温至80±5℃,保温0.5~1小时,灭酶,再将酶解液过1~2μm滤膜,维持温度在55±5℃,取过滤液,过滤液喷雾干燥后获得胶原蛋白肽;
取弹性蛋白肽原料带洗净,加入0.5%NaOH溶液,在40~50℃下浸泡1~2小时,去除表面脂肪及杂质,水洗至中性后沥干,绞碎,得到弹性蛋白粗提物,加入2~4倍量水,调节pH7.5~8.5,加入木瓜蛋白酶和/或复合蛋白酶Protamex,酶的加入量为0.4~0.8%,在60℃保温6~8小时,升温至90℃,保温0.5~1小时,灭酶,采用板框过滤,并过0.45~1μm滤膜,收集滤液,得到弹性蛋白肽溶液,浓缩后,喷粉,得到弹性蛋白肽;
混合胶原蛋白肽和弹性蛋白肽获得所述组合物。
有本发明制备方法制备的胶原蛋白肽经高效体积排阻色谱法测定胶原蛋白分子量为2000Da~6000Da,弹性蛋白肽经高效凝胶排阻色谱法测定弹性蛋白分子量为300Da~2000Da。两者按照本发明配比组合后与单一蛋白肽对比,在同等程度的用量前提下,本发明所述组合物能够显著提高皮肤受损的模型小鼠皮肤中弹性蛋白以及胶原蛋白的含量,并显著提高透明质酸,羟辅氨酸含量和降低MMP3含量;或者说在使用更少的弹性蛋白肽和胶原蛋白肽的前提下,本发明组合物能够达到或超过使用量较多的弹性蛋白肽或胶原蛋白肽的各方面效果;同时,本发明通过不同制备工艺设立对照组,各对照组均无法实现本发明上述优异的效果。
此外,本发明所述组合物还能够在用量较小的前提下,显著抑制皮肤炎症因子,并提高抗氧化酶的含量。
基于上述的优异效果,本发明提出了所述组合物在如下任意一个方面中的应用:
制备美容产品、制备食品和制备药品。
其中,美容产品、食品和药品的剂型包括但不限于固体制剂(如固体饮料、粉剂、胶囊、片剂等)和液体剂型(如口服液、饮料等)等。
由以上技术方案可知,本发明通过特定的工艺制备获得了特定分子量区间的弹性蛋白肽和胶原蛋白肽,两者在合适的比例下组成的组合物,能够以较少的使用量同时显著提高受损皮肤中弹性蛋白和胶原蛋白的含量,并同时显著提高透明质酸,羟辅氨酸含量和降低MMP3含量,同时还能够抑制皮肤炎症因子。
具体实施方式
本发明公开了一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法和应用,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明。本发明所述组合物及其制备方法和应用已经通过实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文所述组合物及其制备方法和应用进行改动或适当变更与组合,来实现和应用本发明技术。
在本发明具体实施例中,涉及到对比试验,各试验组除了应有的区别外,其余试验环境和原材料等均保持一致。
以下就本发明所提供的一种胶原蛋白肽和弹性蛋白肽的组合物及其制备方法和应用做进一步说明。
实施例1:制备本发明所述组合物
取鳕鱼皮经剪碎,置于80℃烘箱烘干12小时,后用粉碎机粉碎成粉末,置于阴凉干燥处备用。称取鳕鱼皮粉,加入20-30倍量水,溶液升温至60℃±5℃,向溶液中加入1%胃蛋白酶,酶解4~8小时,升温至80±5℃,保温0.5~1小时,灭酶,再将酶解液过1μm滤膜,维持温度在55±5℃,取过滤液。过滤液喷雾干燥后获得胶原蛋白肽。经高效体积排阻色谱法测定胶原蛋白分子量为2000Da~6000Da;
取牛韧带洗净,加入0.5%NaOH溶液,在40~50℃下浸泡1~2小时,去除表面脂肪及杂质,水洗至中心后沥干,绞碎,得到弹性蛋白粗提物,加入2~4倍量水,调节pH 7.5~8.5,加入复合蛋白酶Protamex,酶的加入量为0.4~0.8%,在60℃保温6~8小时,升温至90℃,保温0.5~1小时,灭酶,采用板框过滤,并过0.45μm滤膜,收集滤液,得到弹性蛋白肽溶液,浓缩后,喷粉,得到弹性蛋白肽。经高效凝胶排阻色谱法测定弹性蛋白分子量为300Da~2000Da。
将制备的胶原蛋白肽和弹性蛋白肽按照(1.6-3.2):(0.075-0.2)质量比复配成组合物,配比可优选为(1.6-2.4):0.1,如1.6:0.1(3.2g胶原蛋白肽+0.2g弹性蛋白肽)或2.4:0.1(4.8g胶原蛋白肽+0.2g弹性蛋白肽)。
实施例2:动物实验
1、实验材料
D-半乳糖:购自Sigma-Aldrich公司;Mouse Hyaluronic acid(HA)ELISA kit、Mouse hydroxyproline(Hyp)ELISA kit、Mouse Collagen Type I(ColI)ELISA kit、MMP3ELISA kit、Mouse Elastin ELISAkit、IL-1αELISA试剂盒、IL-4试剂盒购自武汉华美生物工程有限公司;GSH-PX购自南京建成生物工程研究所。
2、实验仪器
电子天平:Mettler Toledo,型号:PL303;皮肤水分测定仪:Huntkey便携弹性水分油光皮肤测试仪(HUNTKEY JAPAN株式会社),型号:HKJ-SK03P;紫外灭菌灯:飞利浦黑光紫光管TL-D紫外线杀菌灯(飞利浦照明公司),型号:TL-D/BLB;离心机:Kylin-BellLabInstruments,型号:LX-200;酶标仪:美国BIO-TEK,型号:ELX-800;匀浆机:SONICSMATERIALS INC,型号:VCX130PB;EP管,注射器等。
3、实验动物
BALBc-nu裸小鼠(SPF级,雄性3~4周龄),由中山大学动物实验中心提供,饲养于SPF级屏障动物房。
4、实验方法
4.1实验动物分组及处理模型组每日颈背皮下注射10%D-gal1.0g·kg-1,同时进行UV照射,UV照射波长为350~400nm,照射时间是40min·d-1,光源距鼠约40cm垂直高处,持续造模42d。正常对照组每日颈背部皮下注射等体积的生理盐水,正常光照饲养。实验期间小鼠自由进食和饮水,饮水每天更换。造模同时各样品组喂食各剂量样品干预,喂食前按比例稀释使各组给药体积一致,喂食方法为经口灌胃喂食,每天1次,连续给药42d,正常组和模型对照组用同种喂食方法给予同等容积的蒸馏水。组别设置:空白组、模型组、A单胶原蛋白组(A1:3.2g;A2:6.4g;A3:12.8g),B单弹性蛋白肽组(B1:0.2g;B2:0.4g;B3:0.8g),C胶原蛋白肽+弹性蛋白肽组(C1:1.7g(1.6g+0.1g);C2:3.4g(3.2g+0.2g);C3:6.8g(6.4g+0.4g));
同时,设置3组对照组:
D1组,参照实施例1工艺,区别在于,使用的酶为碱性蛋白酶(用于胶原蛋白肽,1%)和风味蛋白酶(用于弹性蛋白肽,0.8%),胶原蛋白肽和弹性蛋白肽的质量比为1.6:0.1,即3.2g+0.2g;
D2组,参照实施例1工艺,区别在于,更换滤膜的孔径,胶原蛋白肽采用滤径4.0μm滤膜,弹性蛋白肽采用滤径2.0μm滤膜,胶原蛋白肽和弹性蛋白肽的质量比为1.6:0.1,即3.2g+0.2g;
D3组,照专利CN106519020实施例2的工艺,功能性肽用量为3.4g;
4.2皮肤组织I型胶原蛋白、弹性蛋白、透明质酸、羟脯氨酸、MMP3、IL-1α、IL-4、GSH-PX的检测采血后,脱臼颈椎处死小鼠,取1.5cm×1.5cm背部皮肤,取余下背部皮肤用于干燥法测量皮肤含水量和皮肤组织病理学检查,剪去皮下组织称重,4℃0.9%NaCl溶液中漂洗,吸水纸拭干,剪碎,加入4℃0.9%NaCl溶液(0.9%NaCl溶液的体积总量是皮肤重量的9倍)和适量蛋白裂解液,用组织匀浆机匀浆(冰上操作,每次10s,间隔30s,重复5~6次),匀浆用离心机3000r/min冷冻离心15min,取适量上清液按ELISA试剂盒说明检测并计算皮肤组织I型胶原蛋白、弹性蛋白、透明质酸、羟脯氨酸、MMP3、IL-1α、IL-4、GSH-PX活性。
5、统计学分析
实验数据由GraphPad Prism 6.0生物统计学软件进行统计学处理:计量数据采用均数±标准差(mean±SD)表示,采用方差分析结合Dunnett’s多重比较法进行分析;计数资料采用Kruskal-Wallis秩和检验进行分析。
6、结果
(1)胶原蛋白
如表1数据所示,与空白组相比,模型组裸鼠皮肤中I型胶原蛋白含量明显降低(P<0.001);与模型组相比,单用胶原蛋白肽的A3组和单用弹性蛋白肽的B3组有显著的升高皮肤中I型胶原蛋白的作用(P<0.001),但是用量相对较高;而本发明组合物的C2组,用量为A3组+B3组的四分之一,但却相比模型组有显著的升高皮肤中I型胶原蛋白的作用(P<0.01),本发明组合物的C3组,用量为A3组+B3组的二分之一,但却相比模型组有显著的升高皮肤中I型胶原蛋白的作用(P<0.001)。这表明了本发明组合物具备明显的协同增效作用。
同时,三个对照组中,调整酶的类型以及过滤滤膜孔径的D1组和D2组均无法显著提高胶原蛋白的含量;而D3组虽能够显著提高胶原蛋白的含量,但是却无法显著提高弹性蛋白的含量。
表1
组别 | COLLAGEN I(IOD) | ELASTIN(IOD) |
空白组 | 5228±146*** | 5044±141*** |
模型组 | 2777±63 | 2585±242 |
A1 | 2855±39 | 2051±39 |
A2 | 3361±60 | 3088±432 |
A3 | 4774±167*** | 3206±75 |
B1 | 2822±163 | 2749±209 |
B2 | 3373±144 | 3530±254 |
B3 | 3808±94*** | 4360±315** |
C1 | 2887±116 | 3144±23 |
C2 | 3653±205** | 3630±282** |
C3 | 4925±47*** | 4877±64*** |
D1 | 3263±174 | 3430±214 |
D2 | 3423±251 | 3390±152 |
D3 | 3723±209** | 3360±174 |
注:与模型组相比,*P<0.05,**P<0.01,***P<0.001;
(2)弹性蛋白
如表1数据所示,与空白组相比,模型组裸鼠皮肤中弹性蛋白含量明显降低(P<0.001);与模型组相比,单用胶原蛋白肽的A1-3组和单用弹性蛋白肽的B1-2组均不能显著提高裸鼠皮肤中的弹性蛋白含量,特别是单用胶原蛋白肽A1组,反而有下降的趋势;单用弹性蛋白B3组有显著的升高皮肤中弹性蛋白的作用(P<0.001),但是用量相对较高,达到0.8g;而本发明组合物的C3组,用量与A2组+B2组相同,为A3组+B3组的二分之一,但却相比模型组有显著的升高皮肤中弹性蛋白的作用(P<0.001)。这表明了本发明组合物具备明显的协同增效作用。
同时,三个对照组中,调整酶的类型以及过滤滤膜孔径的D1组和D2组均无法显著提高弹性蛋白的含量;而D3组也未能显著提高弹性蛋白的含量。
(3)MMP3,透明质酸,羟辅氨酸
如表2所示,与空白组相比,模型组裸鼠皮肤中MMP3、透明质酸、羟辅氨酸含量形成明显差异(P<0.001);与模型组相比,单用胶原蛋白肽的A1-3组和单用弹性蛋白肽的B1-3组能显著改善裸鼠皮肤中的MMP3、透明质酸、羟辅氨酸含量(P<0.05;P<0.01),但用量较高;而本发明组合物的C2组,用量为A3组+B3组的四分之一,但却相比模型组能显著改善裸鼠皮肤中的MMP3、透明质酸、羟辅氨酸含量(P<0.05),本发明组合物的C3组,用量为A3组+B3组的二分之一,但却相比模型组能显著改善裸鼠皮肤中的MMP3、透明质酸、羟辅氨酸含量(P<0.01;P<0.001)。这表明了本发明组合物具备明显的协同增效作用。
同时,D1-D3组虽能够显著提高透明质酸的含量,但是对于提高羟辅氨酸含量和降低MMP3含量却无法达到预期目的。
表2(ELISA试剂盒检测)
注:与模型组相比,*P<0.05,**P<0.01,***P<0.001;
(4)皮肤炎症因子
如表3所示,与空白组相比,模型组裸鼠皮肤中IL-1α和IL-4炎症因子显著升高,同时GSH-Px明显下降;虽然单用胶原蛋白肽的A1-3组和单用弹性蛋白肽的B1-3组能显著改善上述指标中一个或多个,但是本发明组合物能够在其四分之一或二分之一的用量下,同样达到显著改善的目的,这表明了本发明组合物具备明显的协同增效作用。
同时,D1-D3组只能改善IL-1α炎症因子的指标,对于两外两种指标无法达到改善作用,这明显不如本发明改善多种炎症因子指标的效果。
表3(ELISA试剂盒检测)
组别 | IL-1α(pg/ml) | IL-4(pg/ml) | GSH-Px(ng/L) |
空白组 | 94.67±2.45*** | 255.02±9.31*** | 265.93±2.11*** |
模型组 | 130.41±3.41 | 287.35±3.11 | 194.46±2.17 |
A1 | 129.28±3.29* | 288.60±6.14 | 193.99±1.98 |
A2 | 128.38±5.17* | 279.71±11.21 | 198.88±4.12* |
A3 | 104.94±1.23*** | 256.46±3.27** | 198.68±2.34 |
B1 | 128.81±3.34* | 284.48±6.26 | 195.04±1.45 |
B2 | 117.80±2.43*** | 282.65±5.21 | 194.71±2.67 |
B3 | 96.97±2.43*** | 276.29±4.11** | 199.74±2.21* |
C1 | 129.98±1.18* | 283.96±2.26 | 197.40±3.11* |
C2 | 129.67±3.15* | 287.29±9.11 | 197.46±4.09* |
C3 | 114.15±2.23*** | 279.98±7.25** | 196.68±1.34* |
D1 | 129.28±3.29* | 281.60±4.42 | 192.93±1.87 |
D2 | 129.28±3.29* | 282.60±5.44 | 194.99±1.58 |
D3 | 128.97±2.45* | 283.40±4.12 | 196.44±1.68 |
注:与模型组相比,*P<0.05,**P<0.01,***P<0.001;
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (9)
1.一种胶原蛋白肽和弹性蛋白肽的组合物,其特征在于,由胶原蛋白肽和弹性蛋白肽组成;所述胶原蛋白肽由胶原蛋白原料经胃蛋白酶或胰蛋白酶酶解制成,所述弹性蛋白肽由弹性蛋白原料经木瓜蛋白酶和/或复合蛋白酶Protamex酶解制成。
2.根据权利要求1所述组合物,其特征在于,所述胶原蛋白肽和弹性蛋白肽的质量比为(1.6-3.2):(0.075-0.2)。
3.根据权利要求2所述组合物,其特征在于,所述胶原蛋白肽和弹性蛋白肽的质量比为(1.6-2.4):0.1。
4.根据权利要求1所述组合物,其特征在于,所述胶原蛋白原料为鳕鱼。
5.根据权利要求1所述组合物,其特征在于,所述弹性蛋白原料为牛韧带、牛心管、鲣鱼动脉球和猪心脏大动脉中的一种或两种以上。
6.根据权利要求1-5任意一项所述组合物在如下任意一个方面中的应用:
制备美容产品、制备食品和制备药品。
7.权利要求1所述组合物的制备方法,其特征在于,包括:
胶原蛋白原料加胃蛋白酶或胰蛋白酶酶解,酶解液过1~2μm滤膜,滤液喷雾干燥获得胶原蛋白肽;
弹性蛋白肽原料去除表面脂肪和杂质,加木瓜蛋白酶和/或复合蛋白酶Protamex酶酶解,酶解液过0.45~1μm滤膜,滤液喷粉获得弹性蛋白肽;
混合胶原蛋白肽和弹性蛋白肽获得所述组合物。
8.根据权利要求1所述制备方法,其特征在于,包括:
胶原蛋白原料干燥粉碎,加水以及1%胃蛋白酶或胰蛋白酶酶解,酶解液过1~2μm滤膜,滤液喷雾干燥获得胶原蛋白肽;
弹性蛋白肽原料加NaOH溶液浸泡去除表面脂肪和杂质,清洗并绞碎,加水并调节pH7.5~8.5,加入木瓜蛋白酶和/或复合蛋白酶Protamex酶解,酶的加入量为0.4~0.8%,酶解液采用板框过滤,并过0.45~1μm滤膜,滤液浓缩后喷粉,获得弹性蛋白肽;
混合胶原蛋白肽和弹性蛋白肽获得所述组合物。
9.根据权利要求8所述制备方法,其特征在于,包括:
取胶原蛋白肽原料剪碎,置于80℃烘箱烘干12小时,后用粉碎机粉碎成粉末,加入20-30倍量水,溶液升温至60℃±5℃,向溶液中加入1%胃蛋白酶/胰蛋白酶,酶解4~8小时,升温至80±5℃,保温0.5~1小时,灭酶,再将酶解液过1~2μm滤膜,维持温度在55±5℃,取过滤液,过滤液喷雾干燥后获得胶原蛋白肽;
取弹性蛋白肽原料带洗净,加入0.5%NaOH溶液,在40~50℃下浸泡1~2小时,去除表面脂肪及杂质,水洗至中性后沥干,绞碎,得到弹性蛋白粗提物,加入2~4倍量水,调节pH7.5~8.5,加入木瓜蛋白酶和/或复合蛋白酶Protamex,酶的加入量为0.4~0.8%,在60℃保温6~8小时,升温至90℃,保温0.5~1小时,灭酶,采用板框过滤,并过0.45~1μm滤膜,收集滤液,得到弹性蛋白肽溶液,浓缩后,喷粉,得到弹性蛋白肽;
混合胶原蛋白肽和弹性蛋白肽获得所述组合物。
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