WO2022217490A1 - 一种胶原蛋白肽的制备方法 - Google Patents

一种胶原蛋白肽的制备方法 Download PDF

Info

Publication number
WO2022217490A1
WO2022217490A1 PCT/CN2021/087165 CN2021087165W WO2022217490A1 WO 2022217490 A1 WO2022217490 A1 WO 2022217490A1 CN 2021087165 W CN2021087165 W CN 2021087165W WO 2022217490 A1 WO2022217490 A1 WO 2022217490A1
Authority
WO
WIPO (PCT)
Prior art keywords
collagen
extraction method
stock solution
solution
fish skin
Prior art date
Application number
PCT/CN2021/087165
Other languages
English (en)
French (fr)
Inventor
郭红星
周尽学
Original Assignee
海南三元星生物科技股份有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 海南三元星生物科技股份有限公司 filed Critical 海南三元星生物科技股份有限公司
Priority to PCT/CN2021/087165 priority Critical patent/WO2022217490A1/zh
Priority to JP2021555586A priority patent/JP7401932B2/ja
Publication of WO2022217490A1 publication Critical patent/WO2022217490A1/zh

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2/00—Peptides of undefined number of amino acids; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products

Definitions

  • the invention relates to the technical field of protein extraction, in particular to a method for preparing collagen peptides.
  • Collagen is an extracellular protein composed of two or more amino acids - protein peptides.
  • the absorption of human body is carried out in the form of peptides.
  • the absorption and utilization rate of edible protein peptides can reach 100%.
  • Collagen is the most important part of the extracellular matrix.
  • Collagen peptide is an important part of the bone organic matrix.
  • Collagen peptide has physiological activities such as promoting amino acid absorption and protein synthesis, anti-osteoporosis, lowering blood pressure, and anti-oxidation.
  • Collagen peptide is an extracellular protein. It is a fibrous protein with three peptide chains twisted into a spiral shape. Collagen is the most abundant protein in the human body, accounting for 1/3 of the total body protein.
  • a 60kg adult body has about 3kg of collagen, which is mainly found in human skin, bones, eyes, teeth, tendons, internal organs (including heart, stomach, intestines, blood vessels) and other parts. Its function is to maintain skin and tissue. The shape and structure of organs are also important raw materials for repairing damaged tissues. 70% of human skin is composed of collagen. When collagen is insufficient, not only skin and bones will have problems, but also internal organs will be adversely affected. That is to say, collagen is an important component that is indispensable for maintaining the normal activities of the body. It is also a substance that keeps the body young and prevents aging. In addition, collagen can also prevent diseases, improve physical fitness, and are very helpful for beauty and health.
  • Collagen peptides can be actively and barrier-free absorbed by the human body, without digestion, and can be fully utilized by body tissues.
  • the average molecular weight of marine collagen peptides is smaller, and even consists of only 2-4 amino acids, which is far smaller than ordinary peptides.
  • Dalian Xiaoqin Company selects Antarctic deep-sea fish, uses the skin of marine fish as raw material, uses modern bioengineering technology and enzyme engineering technology to extract, and finally extracts it into marine collagen peptides. Now collagen is slowly entering the field of beauty and skin care.
  • Collagen is a One of the main components of human tissue structure, it is also the most abundant protein in the human body, accounting for about 1/3 of the total human body protein, which is equivalent to 6% of the human body weight. Bone, cartilage, ligament, cornea, various intima, fascia, etc. are the main components to maintain the shape and structure of skin and tissues and organs, and are also important raw materials for repairing damaged tissues.
  • the present invention provides a preparation method of collagen peptide, which solves the problem that the finished product prepared by collagen peptide is not obvious and easy to be oxidized and deteriorated.
  • the invention provides a method for extracting collagen peptides, comprising the following steps:
  • the pH of the collagen acid stock solution is adjusted to 7.0 to obtain a neutral collagen stock solution, which is subjected to enzyme inactivation and centrifugation to obtain a supernatant;
  • the supernatant is decolorized and deodorized, concentrated and dried to obtain collagen peptides.
  • the pretreatment is as follows: drying the fish skin at 50-70° C. until the surface is dry and anhydrous, using scissors to cut each piece of fish skin into 4-6 parts, and then using a fish skin equivalent to 6 times the volume of the dried fish skin. Diethyl ether and petroleum ether were allowed to stand for degreasing for 48 hours, and the solution was changed every 24 hours. After degreasing, rinse with clean water.
  • the pH of the collagen acid stock solution is adjusted to 7.0 with a pH 8.0 NaOH solution.
  • the enzyme inactivation is maintained at a temperature above 95° C. for 10 min.
  • the centrifugation conditions are centrifugation for 15 minutes at a rotational speed of 400 r/min.
  • a NaOH aqueous solution with a pH of 8.0 is added to the collagen acid solution, and the neutral collagen stock solution with a pH of 7.0 is adjusted.
  • the invention utilizes the super strong adsorption capacity of activated carbon to decolorize the neutral collagen stock solution, and at the same time, it can also reduce the fishy smell of the raw material itself.
  • the decolorization and deodorization is to add activated carbon to the supernatant, and decolorize at 40°C for 30min.
  • the concentration is concentrated to a solid content of 10-15% under the conditions of 55° C. and a vacuum degree of 0.09-0.1 MPa.
  • the drying is spray drying
  • the air inlet temperature of the spray drying is 170-185°C
  • the air outlet temperature is not 70-85°C
  • the feed temperature is 20-45°C
  • the sample injection speed is 10-20r/ min.
  • the temperature control instrument for pretreatment adopts SC-20A numerical control super constant temperature tank
  • the equipment adopted for the pretreatment oven is electric heating blast drying oven
  • the equipment for measuring pH value in enzymolysis is STARTE2100 laboratory pH meter
  • the centrifugal The processing equipment is TD5A-WS centrifuge
  • the spray drying equipment is DC-1500 laboratory spray dryer.
  • the present invention also provides the collagen peptide prepared by the above preparation method.
  • the invention provides a preparation method of collagen peptide, comprising the following steps: adding 0.5monl/L glacial acetic acid solution and 3% pepsin by mass fraction after pretreatment of fish skin and fish meat, stirring, and enzymatic hydrolysis below 10°C 48h, the collagen acid stock solution was obtained; NaOH solution was added to the collagen acid stock solution to obtain a neutral collagen stock solution, and the supernatant was obtained by inactivating the enzyme and centrifugation; the supernatant was decolorized and deodorized, concentrated and dried to obtain Collagen Peptides.
  • the present invention has the following beneficial effects:
  • the invention uses 0.5monl/L glacial acetic acid solution and 3% pepsin to extract collagen peptides in fish skin and fish meat for enzymatic hydrolysis, which can effectively remove the fat contained in the fish skin and fish meat, so that the fat can be removed.
  • the protein is easily exposed to contact with the enzyme, which is beneficial to the hydrolysis of the substrate, and at the same time improves the color and flavor of the collagen peptide, so that the activity of the collagen peptide is higher, and the process cost is low.
  • it also has the advantages that the reaction temperature and value are relatively mild, and the hydrolysis process is easy to control.
  • Figure 1 is a schematic diagram of the preparation process of collagen peptides.
  • the invention provides a preparation method of collagen peptide.
  • Those skilled in the art can learn from the content of this document and appropriately improve the process parameters to achieve. It should be particularly pointed out that all similar substitutions and modifications will be apparent to those skilled in the art, and they are deemed to be included in the present invention.
  • the method and application of the present invention have been described through the preferred embodiments, and it is obvious that relevant persons can make changes or appropriate changes and combinations of the methods and applications herein without departing from the content, spirit and scope of the present invention, so as to realize and apply the present invention. Invention technology.
  • test materials used in the present invention are all common commercial products and can be purchased in the market.
  • Sp1 raw material
  • the raw material is fresh fish skin to remove the fish lin and adhering fish meat. After washing and sub-packaging, put it in a -18 °C refrigerator for storage.
  • Fish skin and fish scales are ideal raw materials for preparing blood pressure-lowering active peptides.
  • people's demand for collagen from aquatic animals is getting higher and higher.
  • diseases such as mad cow disease, foot-and-mouth disease, and avian influenza
  • the big question is the limited application of terrestrial animal-derived collagen in Muslim areas.
  • the pretreatment is to put the thawed fish skin into an oven, dry and anhydrous, and then carry out degreasing treatment to obtain degreasing fish skin and fish meat, which are placed in a -18°C refrigerator for refrigeration for later use; wherein, the pretreatment is Put the thawed fish skin into the oven, bake the fish skin at a temperature of 50-70 °C until the surface is dry and no water, use scissors to cut each piece of fish skin into 4-6 pieces, and then use the equivalent of dried fish skin, 6 times the volume of ether and petroleum ether were left to degrease for 48h, and the solution was changed every 24h. After degreasing, the remaining ether and petroleum ether in the fish skin were washed with water, and placed in a -18°C refrigerator for later use.
  • Enzymatic hydrolysis the enzymatic hydrolysis process is to take out the defatted fish meat and skin from the refrigerator, first add 1 part of defatted fish meat to 30 times of 0.5monl/L glacial acetic acid solution and 1 part of 3% pepsin Stirring treatment, enzymatic hydrolysis below 10 °C for 48 h, to obtain collagen acid stock solution,
  • inactivation of enzymes is to heat the neutral collagen stock solution to above 95°C and keep it for 10min after enzymolysis, so as to achieve the effect of fully inactivation of enzymes;
  • the neutral collagen stock solution adopts the TD5A-WS type centrifuge to centrifuge the enzyme-killing liquid after the enzymolysis is finished, and the centrifugation condition is centrifugal 15min under the condition of rotating speed 400r/min;
  • the decolorization condition is to add activated carbon with a mass fraction of 0.75% to the neutral collagen stock solution, and decolorize it for 30 minutes at a temperature of 40 °C. In order to obtain products with higher quality and stronger activity in the production of collagen peptides, it is necessary to After activated carbon decolorization, membrane filtration and other treatments;
  • Sp7 Concentrating, concentrating the neutral collagen stock solution through a rotary evaporator at a temperature of 55° C. and a vacuum degree of 0.09-0.1 MPa, and controlling the solid content of the concentrated solution to be about 10-15% to obtain a concentrated solution;
  • the spray drying adopts a pressure type spray dryer, the inlet air temperature is set to 170-185°C, the outlet air temperature is 70-85°C, the feed temperature is 20-45°C, and the sampling speed is 10-20r/min , the obtained concentrated solution is dried to obtain a white powdery crude peptide;
  • the analysis and determination is that the moisture content of the crude peptide is determined by the constant weight method at 105°C, the crude fat content of the crude peptide is determined by the Soxhlet extraction method, and the calcination constant weight method at 550°C is used for the determination of the crude peptide.
  • the ash content of the crude peptide was determined, and the protein content of the crude peptide was determined by the Kjeldahl method.
  • M the total amount of protein before the amount of activated carbon treatment
  • M the total protein mass after treatment with activated carbon
  • Acid standard working solution 1.0, 2.5, 5.0, 7.5, 10.0mL, respectively, use hydrochloric acid to dilute to 100mL, the concentration is 1.0, 2.5, 5.0, 7.5, 10.0ug/mL, each take 1mL of 0.001mol/L hydrochloric acid
  • the solution was used as a blank solution, and 2 mL of chloramine solution was added respectively, and allowed to stand at room temperature for 20 minutes, then 2 mL of perchloric acid solution was added, and 2 mL of perchloric acid solution was added, and 2 mL of p-dimethylaminobenzoic acid solution was added.

Abstract

一种胶原蛋白肽的制备方法，包括如下步骤：将鱼皮和鱼肉预处理后加入0.5monl/L的冰醋酸溶液和质量分数为3％的胃蛋白酶搅拌，10℃以下酶解48h，得到胶原酸原液；调节所述胶原酸原液pH至7.0，得到中性胶原原液，经灭酶、离心，得到上清液；将所述上清液脱色脱腥、浓缩、干燥，获得胶原蛋白肽。

Description

一种胶原蛋白肽的制备方法 技术领域
本发明涉及蛋白提取技术领域，尤其涉及一种胶原蛋白肽的制备方法。
背景技术
胶原蛋白是一种细胞外蛋白质，由二个或二个以上氨基酸组成－－蛋白肽。人体的吸收是以肽的方式进行，食用蛋白肽的吸收利用率可达100％，胶原蛋白是细胞外基质中最重要的组成部分，胶原蛋白肽是骨有机基质的重要组成部分，其水解产物胶原肽具有促进氨基酸吸收和蛋白质合成、抗骨质疏松、降血压、抗氧化等生理活性，胶原蛋白肽是一种细胞外蛋白质，它是曲3条肽链拧成螺旋形的纤维状蛋白质，胶原蛋白是人体内含量最丰富的蛋白质，占全身总蛋白质的1/3。一个60kg的成年人的身体内约有3公斤胶原蛋白，主要存在于人体皮肤、骨骼、眼睛、牙齿、肌腱、内脏(包括心、胃、肠、血管)等部位，其功能是维持皮肤和组织器官的形态和结构，也是修复各损伤组织的重要原料物质。在人体皮肤成分中，有70％是由胶原蛋白所组成，当胶原蛋白不足时，不仅皮肤及骨骼会出现问题，对内脏器官也会产生不利影响。也就是说，胶原蛋白是维持身体正常活动所不可缺少的重要成分。同时也是使身体保持年轻、防止老化的物质。另外，胶原蛋白还可以预防疾病，改善体质，对美容和健康都很有帮助，胶原蛋白肽，可以被人体主动且无障碍吸收，无需消化，供身体组织充分利用。并且海洋胶原肽平均分子量更小，甚至只有2-4个氨基酸构成，远远小于普通肽。大连晓芹公司精选南极深海海鱼，以海鱼的鱼皮为原料，运用现代生物工程技术及酶工程技术提取，最后提炼为海洋胶原肽，现在胶原蛋白正慢慢进入美容护肤领域.胶原蛋白是人体组织结构的主要成分之一，也是人体内含量最多的一种蛋白质，约占人体蛋白质总量的1/3，相当于人体体重的6％，它遍及全身的各个组织器官，如：皮肤、骨骼、软骨、韧带、角膜、各种内膜、筋膜等，是维持皮肤与组织 器官形态、结构的主要成分，也是修复各损伤组织的重要原料物质。
就目前对加元蛋白肽的提取制备工艺在实际使用时，由于提取原料鱼类，动物类，等不同提取原料的不同需要采取相应的提取分解工艺，同时在提取胶原蛋白肽时容易忽略原料中脂肪的取出，进而使得到的胶原蛋白肽容易氧化变质，影响产品的色泽和风味，脂肪的寸在容易使胶原蛋白肽中蛋白质被包裹在脂肪内，不利于水解使用，从而使胶原蛋白肽的功效大大降低。
发明内容
针对现有技术的不足，本发明提供了一种胶原蛋白肽的制备方法，解决了胶原蛋白肽制备成品功效不明显易氧化碳变质等问题。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供一种胶原蛋白肽的提取方法，包括如下步骤：
将鱼皮和鱼肉预处理后加入0.5monl/L的冰醋酸溶液和质量分数为3％的胃蛋白酶搅拌，10℃以下酶解48h，得到胶原酸原液；
调节所述胶原酸原液pH至7.0，得到中性胶原原液，经灭酶、离心，得到上清液；
将所述上清液脱色脱腥、浓缩、干燥，获得胶原蛋白肽。
作为优选，所述预处理为：在50-70℃下将鱼皮烘至表面干燥无水，用剪刀将每块鱼皮剪成4-6份，再用相当于鱼皮干6倍体积的乙醚、石油醚静置脱脂48h，每24h更换溶液1次,脱脂结束后用清水洗净。
作为优选，利用pH8.0的NaOH溶液调节所述胶原酸原液pH至7.0。
作为优选，所述灭酶为95℃以上保持10min。
作为优选，所述离心条件为转速400r/min条件下离心15min。
本发明提供的制备方法中，向胶原酸溶液中加入PH为8.0的NaOH水溶液，调至PH为7.0的中性胶原原液。
本发明利用活性炭的超强吸附能力，对中性胶原原液进行脱色，同时也可减弱原料本身的腥味。
作为优选，所述脱色脱腥为在所述上清液中加入活性炭，40℃下脱 色30min。
作为优选，所述浓缩为55℃、0.09-0.1MPa真空度的条件下浓缩至固形物含量为10-15％。
作为优选，所述干燥为喷雾干燥，所述喷雾干燥的进风温度为170-185℃，出风温度未70-85℃，进料温度为20-45℃，进样速度为10-20r/min。
本发明中，预处理的温度控制仪器采用SC-20A数控超级恒温槽，预处理的烘箱采用的设备为电热鼓风干燥箱，酶解中测定PH值的设备为STARTE2100型实验室PH计，离心处理采用设备为TD5A-WS型离心机，喷雾干燥设备为DC-1500型实验室型喷雾干燥机。
本发明还提供了由上述制备方法制得的胶原蛋白肽。
本发明提供了一种胶原蛋白肽的制备方法，包括如下步骤：将鱼皮和鱼肉预处理后加入0.5monl/L的冰醋酸溶液和质量分数为3％的胃蛋白酶搅拌，10℃以下酶解48h，得到胶原酸原液；向所述胶原酸原液中加入NaOH溶液，得到中性胶原原液，经灭酶、离心，得到上清液；将所述上清液脱色脱腥、浓缩、干燥，获得胶原蛋白肽。本发明具有以下有益效果：
本发明利用0.5monl/L的冰醋酸溶液和质量分数为3％的胃蛋白酶提取鱼皮和鱼肉中的胶原蛋白肽进行酶解，可有效去除鱼皮和鱼肉中所含的脂肪，使去除脂肪的蛋白质跟容易暴露出来与酶进行接触，有利于底物的水解，同时提高胶原蛋白肽的色泽和风味，使胶原肽活性更高，且工艺成本低。同时还具有反应温度和值较为温和，水解过程易于控制的优点。
附图说明
图1为胶原蛋白肽的制备流程示意图。
具体实施方式
本发明提供了一种胶原蛋白肽的制备方法。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。本发明的方法及应用已经通过较佳实施例进行了描述，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的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。
本发明采用的试材皆为普通市售品，皆可于市场购得。
下面结合实施例，进一步阐述本发明：
实施例1
本发明胶原蛋白肽的制备工艺如图1所示，具体包括如下步骤：
Sp1：原料，原料为新鲜鱼皮去除鱼麟和粘附的鱼肉，洗净分装后放入-18℃的冰箱中保存备用，鱼皮、鱼鳞等，是制备降血压活性肽的理想原料，采用鱼肉为原料，人们对水生动物来源的胶原蛋白的需求越来越高，一方面是由于疯牛病、口蹄疫、禽流感等疾病的发生，使陆生动物胶原蛋白及其制品的安全性受到了极大的质疑，另一方面是陆生动物来源的胶原蛋白在穆斯林地区应用受限。研究表明，水生动物来源的胶原蛋白的某些性质较陆生动物来源的胶原蛋白具有优势，如氨基酸组成无显著差异、低抗原性、低变性温度、易被蛋白酶水解及具有一定的凝胶性、高度分散性及乳化性，更利于胶原蛋白肽的提取，使提取的胶原蛋白肽含量更高，；
Sp2：预处理，预处理为将解冻后的鱼皮放入烘箱，干燥无水处理后，进行脱脂处理，得到脱脂鱼皮和鱼肉，放入-18℃的冰箱冷藏备用；其中，预处理为将解冻后的鱼皮放入烘箱，在50-70℃的温度下将鱼皮烘至表面干燥无水，用剪刀将每块鱼皮剪成4-6份，再用相当于鱼皮干,6倍体积的乙醚、石油醚静置脱脂48h，每24h更换溶液1次,脱脂完后用清水洗净鱼皮中残留的乙醚、石油醚，放入-18℃的冰箱冷藏备用。
Sp3：酶解，酶解过程为将脱脂后的鱼肉和鱼皮从冰箱取出，先以1份脱脂鱼肉加以30倍的0.5monl/L的冰醋酸溶液和1份质量分数为3％的胃蛋白酶搅拌处理，10℃以下酶解48h，得到胶原酸原液，
Sp4：灭酶，灭酶为将酶解过后将中性胶原原液加热至95℃以上并保持10min，以达到充分灭酶的效果；
Sp5：离心，酶解结束后中性胶原原液采用TD5A-WS型离心机将灭 酶液体离心处理，离心条件为转速400r/min条件下离心15min；
Sp6：脱色，脱色条件为对中性胶原原液内加入质量分数为0.75％的活性炭，在温度为40℃的环境下脱色30min，胶原肽生产中为了获取品质更高、活性更强的产品，需经过活性炭脱色、膜过滤等处理；
Sp7：浓缩，浓缩为将中性胶原原液通过旋转蒸发器在温度55℃，真空度0.09-0.1MPa的条件下进行浓缩，控制浓缩液的固形物含量约为10-15％，得到浓缩液；
Sp8：喷雾干燥，喷雾干燥为釆用压力式喷雾干燥机，设置进风温度170-185℃，出风温度70-85℃，进料温度为20-45℃，进样速度10-20r/min，对得到的浓缩液进行干燥，得到白色粉末状粗肽；
Sp9：分析测定，分析测定为分别采用105读C恒重法对粗肽的水分含量进行测定，采用索氏抽提法对粗肽的粗脂肪含量进行测定，采用550℃灼烧恒重法对粗肽的灰分进行测定，采用凯氏定氨法对粗肽的蛋白含量进行测定。
分析测定时，蛋白质的保留率计算公式1-(M前-M后)/M前*100，M前为数量活性炭处理前蛋白质总量，M后为与活性炭处理后蛋白质总质量，测定粗肽中加元蛋白含量时，首先需要绘制羟脯氨酸标准曲线，精确称取羟脯氨酸标准品若干，用的0.001mol/L的盐酸溶解，配制100ug/mL的标准溶液，吸取羟脯氨酸标准工作液1.0、2.5、5.0、7.5、10.0mL，分别用的盐酸定容至100mL，浓度分别为1.0、2.5、5.0、7.5、10.0ug/mL,各取1mL以0.001mol/L的盐酸溶液作为空白液，分别加入2mL的氯胺溶液，室温下静置20min，再加入2mL的高氯酸溶液，室温下静置5min，最后加入2mL对二甲基氨基苯甲酸溶液，60℃加热显色20min，冷却，以空白液进行调零，测定溶液在560nm处的吸光度,采用FA2004电子天平精确称取0.01g样品于安培瓶中，加入6mol/L的盐酸1mL，取1mL溶液置于130℃烘箱总水解3h,冷却后，蒸馏水定容至100mL，滤纸过滤，取1mL按羟脯氨酸标准曲线的测定方法进行测定，将测得的羟脯氨酸标准曲线含量再乘以转换系数13.3，即可得胶原蛋白含量。
需要说明的是，在本文中，诸如第一和第二等之类的关系术语仅仅用来将一个实体或者操作与另一个实体或操作区分开来，而不一定要求或者暗示这些实体或操作之间存在任何这种实际的关系或者顺序。而且，术语“包括”、“包含”或者其任何其他变体意在涵盖非排他性的包含，从而使得包括一系列要素的过程、方法、物品或者设备不仅包括那些要素，而且还包括没有明确列出的其他要素，或者是还包括为这种过程、方法、物品或者设备所固有的要素。在没有更多限制的情况下，由语句“包括一个引用结构”限定的要素，并不排除在包括所述要素的过程、方法、物品或者设备中还存在另外的相同要素。
尽管已经示出和描述了本发明的实施例，对于本领域的普通技术人员而言，可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由所附权利要求及其等同物限定。

Claims (9)

  1. 一种胶原蛋白肽的提取方法，其特征在于，包括如下步骤：
    将鱼皮和鱼肉预处理后加入0.5monl/L的冰醋酸溶液和质量分数为3％的胃蛋白酶搅拌，10℃以下酶解48h，得到胶原酸原液；
    调节所述胶原酸原液pH至7.0，得到中性胶原原液，经灭酶、离心，得到上清液；
    将所述上清液脱色脱腥、浓缩、干燥，获得胶原蛋白肽。
  2. 根据权利要求1所述的提取方法，其特征在于，所述预处理为：在50-70℃下将鱼皮烘至表面干燥无水，用剪刀将每块鱼皮剪成4-6份，再用相当于鱼皮干6倍体积的乙醚、石油醚静置脱脂48h，每24h更换溶液1次,脱脂结束后用清水洗净。
  3. 根据权利要求1所述的提取方法，其特征在于，利用pH8.0的NaOH溶液调节所述胶原酸原液pH至7.0。
  4. 根据权利要求1所述的提取方法，其特征在于，所述灭酶为95℃以上保持10min。
  5. 根据权利要求1所述的提取方法，其特征在于，所述离心条件为转速400r/min条件下离心15min。
  6. 根据权利要求2所述的提取方法，其特征在于，所述脱色脱腥为在所述上清液中加入活性炭，40℃下脱色30min。
  7. 根据权利要求1所述的提取方法，其特征在于，所述浓缩为55℃、0.09-0.1MPa真空度的条件下浓缩至固形物含量为10-15％。
  8. 根据权利要求2所述的提取方法，其特征在于，所述干燥为喷雾干燥，所述喷雾干燥的进风温度为170-185℃，出风温度未70-85℃，进料温度为20-45℃，进样速度为10-20r/min。
  9. 权利要求1～7任一项所述的提取方法制得的胶原蛋白肽。
PCT/CN2021/087165 2021-04-14 2021-04-14 一种胶原蛋白肽的制备方法 WO2022217490A1 (zh)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/CN2021/087165 WO2022217490A1 (zh) 2021-04-14 2021-04-14 一种胶原蛋白肽的制备方法
JP2021555586A JP7401932B2 (ja) 2021-04-14 2021-04-14 コラーゲンペプチドの製造方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2021/087165 WO2022217490A1 (zh) 2021-04-14 2021-04-14 一种胶原蛋白肽的制备方法

Publications (1)

Publication Number Publication Date
WO2022217490A1 true WO2022217490A1 (zh) 2022-10-20

Family

ID=83639935

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/087165 WO2022217490A1 (zh) 2021-04-14 2021-04-14 一种胶原蛋白肽的制备方法

Country Status (2)

Country Link
JP (1) JP7401932B2 (zh)
WO (1) WO2022217490A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116622436A (zh) * 2023-07-25 2023-08-22 烟台海润嘉生物科技有限公司 一种同步提取金枪鱼鱼油和胶原蛋白肽的生产方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101481723A (zh) * 2009-01-23 2009-07-15 熊新安 一种用鲤鱼鱼皮制备胶原蛋白肽的方法
CN101693741A (zh) * 2009-09-05 2010-04-14 泰祥集团技术开发有限公司 一种海洋活性胶原蛋白肽酶解液的脱色脱腥方法
CN103849670A (zh) * 2013-10-22 2014-06-11 中国海洋大学 一种水解鮟鱇鱼皮制备高f值胶原蛋白肽的方法
CN110128530A (zh) * 2019-05-31 2019-08-16 湖北清江鲟龙渔业有限公司 利用鲟鱼原料提取胶原蛋白的工艺

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101979654A (zh) 2010-10-20 2011-02-23 江西科技师范学院 一种从鱼皮中提取胶原蛋白的方法
CN104073536A (zh) 2013-03-25 2014-10-01 林琳 一种多手段协同提取大分子活性胶原蛋白的方法
CN104324367A (zh) 2014-10-20 2015-02-04 广西壮族自治区花红药业股份有限公司 鱼鳞提取物在制备治疗和/或预防白癜风药物方面的新用途
CN105316382A (zh) 2015-12-15 2016-02-10 李国深 一种鱼骨胶原蛋白的制备方法
CN107881208A (zh) 2017-10-09 2018-04-06 浙江海洋大学 一种对非酒精性脂肪肝模型具有修复作用的鳕鱼皮胶原蛋白肽的制备方法
CN108192939A (zh) 2018-01-18 2018-06-22 浙江海洋大学 一种日本黄菇鱼鱼皮胶原蛋白的制备方法
CN109337949A (zh) 2018-10-23 2019-02-15 兰溪市哥特生物技术有限公司 一种深海鱼皮胶原蛋白肽
CN112608968B (zh) 2021-01-14 2022-03-18 山东恒鑫生物科技有限公司 一种以罗非鱼鳞原料生产鱼胶原蛋白肽的方法

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101481723A (zh) * 2009-01-23 2009-07-15 熊新安 一种用鲤鱼鱼皮制备胶原蛋白肽的方法
CN101693741A (zh) * 2009-09-05 2010-04-14 泰祥集团技术开发有限公司 一种海洋活性胶原蛋白肽酶解液的脱色脱腥方法
CN103849670A (zh) * 2013-10-22 2014-06-11 中国海洋大学 一种水解鮟鱇鱼皮制备高f值胶原蛋白肽的方法
CN110128530A (zh) * 2019-05-31 2019-08-16 湖北清江鲟龙渔业有限公司 利用鲟鱼原料提取胶原蛋白的工艺

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116622436A (zh) * 2023-07-25 2023-08-22 烟台海润嘉生物科技有限公司 一种同步提取金枪鱼鱼油和胶原蛋白肽的生产方法

Also Published As

Publication number Publication date
JP7401932B2 (ja) 2023-12-20
JP2023530376A (ja) 2023-07-18

Similar Documents

Publication Publication Date Title
Cho et al. Extracting optimization and physical properties of yellowfin tuna (Thunnus albacares) skin gelatin compared to mammalian gelatins
CN101787078B (zh) 一种胶原蛋白多肽及其制备方法和用途
CN102702346B (zh) 一种从鱼皮中提取酸溶性胶原蛋白的方法及所得到的产品
CN109180808B (zh) 一种鱼鳞胶原蛋白及其制备方法和应用
CN106359631B (zh) 一种纯胶原蛋白奶粉及其制备方法
CN102718862A (zh) 深海鱼类胶原肽的脱色脱腥方法
CN107164439A (zh) 一种适用于化妆品的鱼鳞ⅰ型胶原蛋白肽的规模化制备方法
WO2022217490A1 (zh) 一种胶原蛋白肽的制备方法
CN106983094A (zh) 一种盐渍海蜇的脱铝加工方法
CN109400700A (zh) 一种易吸收的胶原蛋白低聚肽的制备方法
CN107114793A (zh) 一种综合利用鲟鱼骨制备钙和硫酸软骨素的方法
WO2023040084A1 (zh) 胶原三肽的提取方法及制备的胶原三肽
CN113925165B (zh) 脱水桑叶及其加工方法
CN107897941A (zh) 一种胶原蛋白肽片的生产工艺
CN113336842A (zh) 一种胶原蛋白肽的制备方法
CN109293766A (zh) 从鱼鳞中提取胶原蛋白多肽的方法
WO2010043073A1 (zh) 一种胶原多肽及其制备方法和应用
CN102952837A (zh) 一种医用鱼胶原蛋白多肽原料及其制备工艺
CN109275917A (zh) 一种含有海洋鱼皮胶原低聚肽的多肽组合物
CN113122604A (zh) 一种海肠胶及其制备方法和应用
CN106579064A (zh) 一种增强铁吸收的鱼肉休闲食品及其制备方法
CN108048513A (zh) 一种胶原蛋白酶联合新型微生物蛋白酶水解制备乌参胶原蛋白低聚肽的方法
CN102268084A (zh) 一种软黄金胶原蛋白的制备方法和功能研究
CN108277250A (zh) 一种利用鱼骨制备胶原蛋白的方法
CN108865631A (zh) 一种保健醋的制备方法

Legal Events

Date Code Title Description
ENP Entry into the national phase

Ref document number: 2021555586

Country of ref document: JP

Kind code of ref document: A

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21936383

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21936383

Country of ref document: EP

Kind code of ref document: A1