CN102228125A - Preparation method of algal active peptide - Google Patents

Preparation method of algal active peptide Download PDF

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Publication number
CN102228125A
CN102228125A CN2011101404441A CN201110140444A CN102228125A CN 102228125 A CN102228125 A CN 102228125A CN 2011101404441 A CN2011101404441 A CN 2011101404441A CN 201110140444 A CN201110140444 A CN 201110140444A CN 102228125 A CN102228125 A CN 102228125A
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marine alga
hydrolysis
enzyme
active peptide
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CN102228125B (en
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俞继荣
胡汉清
杨红伟
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HUZHOU JIAMEI BIOCHEMICAL PRODUCTS CO Ltd
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Abstract

The invention relates to a preparation method of algal active peptide. The preparation method comprises the following steps of 1) carrying out a coupling catalytic hydrolysis process on food-grade sodium alginate as a raw material by a composite enzyme, wherein key technological parameters of enzyme dosage, PH value, hydrolysis temperature, hydrolysis time and the like are controlled during the hydrolysis process so that a hydrolysis efficiency of the composite enzyme, and carrying out the processes of inactivation, decolorization and filtration on hydrolysate obtained by the hydrolysis process to obtain algal peptide extract, and 2) carrying out the processes of purification and condensation on the algal peptide extract obtained form the step 1 through a nanofiltration membrane with a molecular weight of 1000 Da and a nanofiltration membrane with a molecular weight of 300 Da to obtain algal active products with a molecular weight in a range of 300 to 1000 Da, and carrying out the processes of sterilization and vacuum desiccation on the algal active products to obtain algal active finished products. The preparation method has the advantages of high utilization rate of raw materials, high product purity, no extra process such as a desalination process, a vacuum condensation process and the like, simple production processes and low production cost. The preparation method belongs to the technical field of natural active substance extraction.

Description

The preparation method of marine alga active peptide
Technical field
The present invention relates to a kind of preparation method of marine alga active peptide, belong to natural active matter extractive technique field.
Background technology
Modern nutriology discovers that the mankind ingest protein after gastral enzyme effect, digests and assimilates with low peptide form mostly, and the ratio that absorbs with the free amino acid form is very little.China's peptide paddy expert group further test disclosed again the digestion of peptide specific ionization amino acid faster, absorb manyly, show that the biological value of peptide and nutritive value specific ionization amino acid are higher.
Biologically active peptide is that 25 natural amino acids are formed with difference and the general name of the different peptide classes of the linearity from the dipeptides to the complexity of arrangement mode formation, loop configuration in the protein, is the multi-functional compounds that comes from protein.Biologically active peptide has multiple body metabolism and physiological regulation function, easily digest and assimilate, effects such as promoting immunity, hormone are regulated, antibiotic, antiviral, hypotensive, reducing blood lipid is arranged, edible safety is high, is the most popular research topic of current international food circle and the function factor that has development prospect.
Research is both at home and abroad at present used maximum plant polypeptides and should be belonged to Soybean Peptide, soya-bean polypeptides has the incomparable advantage of protein, as good water-solubility, heat endurance, easily digest and assimilate, low antigenicity, reduction cholesterol and blood pressure, adjusting human physiological functions etc., be applied to fields such as food, medical science, daily-use chemical industry.
Marine alga is rich in multiple life active compound; as polysaccharide, high unsaturated fatty acid, no matter taurine, carotenoid, sterol and laminine etc. are as daily food; still extract active material as medicine, marine alga can both provide very abundant high-grade nutrient element to the human life.Marine algae extract still is confined to algin, alginic acid and algal polysaccharides series products at present, and economic benefit is low, and resource utilization is low.
The marine alga active component of domestic and international similar industry production is mainly sodium alginate now, molecular formula is (C6H7O8Na) n, rely on 1 by α-L-mannuronic acid (M unit) and β-D-guluronic acid (G unit), connection of 4-glycosidic bond and the copolymer of forming by GM, MM and the GG fragment of different proportion.Sodium alginate is slightly soluble in water, is insoluble to most of organic solvent.It is dissolved in alkaline solution, makes solution have viscosity.Sodium alginate powder is met water and is become wet, and the aquation of particulate makes its surface have viscosity.The particulate formation agglomerate that is bonded together rapidly then, agglomerate is aquation and dissolving fully very slowly.If contain the compound of other and alginate competition hydration in the water, then sodium alginate is more difficult is dissolved in the water.Sugar in the water, starch or protein can reduce the hydration rate of sodium alginate.The salt of univalent cation is higher than at 0.5% o'clock in concentration and also has similar effect.
The at present domestic vegetable protein extracting method that relates to has being used in combination of salt and alkali method, acid system, alkaline process and enzyme process and acid, enzyme, being used in combination of alkali, enzyme.Briefly can be divided into chemical degradation method and enzyme edman degradation Edman two classes.Chemical method is the small-molecule substance that breaks to form that impels protein molecule with chemical reagent such as acid, alkali at a certain temperature.Acid system adopts strong acid such as hydrochloric acid, sulfuric acid at high temperature to react more, strong reaction, and equipment corrosion is serious, and hydrolysis is thorough, generates ispol more, and at high temperature tryptophan is destroyed fully simultaneously.Be difficult for being absorbed, gradually be eliminated at present; Alkali process hydrolysis makes the most racemization of amino acid, as makes L-amino acid form D-amino acid, can also form noxious material.To the eighties in 20th century, fast development along with enzyme preparation industry and food industry, it is found that the enzymatic hydrolysis reaction temperature is low, reaction time is short, non-environmental-pollution, and product is of high nutritive value, based on polypeptide and L-type free amino acid, instant capacity is good, is easy to human consumption and absorbs, thereby attention has focused on and utilize on the enzymic hydrolysates albumen.Extract as soya-bean polypeptides, papain, compound protease and hay bacillus etc. are adopted in present domestic enzyme hydrolysis, but, this extraction is undesirable to product control, molecular weight is only accounting for about in the of 35% between the 300-2000Da, active peptides has only about in the of 65%, and free aminoacid content has a long way to go with international most advanced level up to 8%.
Summary of the invention
The objective of the invention is the preparation method that provides a kind of marine alga active peptide for solving the problems of the technologies described above.
Above-mentioned technical purpose of the present invention is achieved by the following technical programs:
The preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, described complex enzyme is that plant rennet and animal protease are with (the complex enzyme that the ratio of 2~4) ︰ 1 mixes; The controlled condition of described catalyzing hydrolysis is that (1~4) ︰ 1, pH are 5~7, and the catalysis time is 120~240min, and temperature is 40~60 ℃ for the enzyme dosage.
Technology point of the present invention is, complex enzyme and use the control of the catalytic condition of this complex enzyme.In the technique scheme of the present invention, the enzyme dosage is meant that enzyme liquid amasss the volume ratio with the substrate sodium alginate soln.The present invention has also adopted the NF membrane technology, this makes the purity height of final products, molecular weight is little, molecular weight 500~3000 daltonian peptide contents reach more than 92%, molecular weight 〉=3000 dalton are in 8%, and human body absorbs easily, and absorptivity reaches more than 90%, has higher purity than like product on the market, higher biologically active.
Preferred as technique scheme, described plant rennet is one or both in bromelain and the papain; Described animal protease is one or both in trypsase, the pepsin.
Preferred as technique scheme, step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 4 ︰ 1; The controlled condition of described catalyzing hydrolysis is 4 ︰ 1 for the enzyme dosage, and pH is 6, and the catalysis time is 240min, and temperature is 40 ℃.
In sum, the present invention has following beneficial effect:
1, the present invention handles marine alga by new bio enzymolysis process technology, under the prerequisite that keeps the original performance of marine alga active peptide, reduces the viscosity of product greatly, has strengthened the compatibility of product in aqua product;
2, the present invention has determined the kind as the new bio enzyme of marine alga enzyme digestion reaction, and has determined its consumption and enzymatic hydrolysis condition to have improved raw material availability greatly;
3, the present invention proposes the brand-new scheme of carrying out nanofiltration after the reaction, improved product purity greatly; Also reach desalination, separation, purifying and concentrated effect simultaneously, simplified production technology;
4, the content that uses the present invention to invent the marine alga active peptide in the product that makes improves greatly, reaches external like product level.
The specific embodiment
This specific embodiment only is an explanation of the invention; it is not a limitation of the present invention; those skilled in the art can make the modification that does not have creative contribution to present embodiment as required after reading this specification, but as long as all are subjected to the protection of Patent Law in claim scope of the present invention.
Embodiment one
The preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 2 ︰ 1; The controlled condition of described catalyzing hydrolysis is 1 ︰ 1 for the enzyme dosage, and pH is 5, and the catalysis time is 120min, and temperature is 60 ℃;
Described plant rennet is a bromelain; Described animal protease is a trypsase.
Embodiment two
The preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 3 ︰ 1; The controlled condition of described catalyzing hydrolysis is 2 ︰ 1 for the enzyme dosage, and pH is 5.5, and the catalysis time is 160min, and temperature is 55 ℃;
Described plant rennet is a papain; Described animal protease is a pepsin.
Embodiment three
The preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 3 ︰ 1; The controlled condition of described catalyzing hydrolysis is 3 ︰ 1 for the enzyme dosage, and pH is 6, and the catalysis time is 200min, and temperature is 50 ℃;
Described plant rennet is the mixture of bromelain and papain; Described animal protease is a pepsin.
Embodiment four
The preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 4 ︰ 1; The controlled condition of described catalyzing hydrolysis is 4 ︰ 1 for the enzyme dosage, and pH is 6.5, and the catalysis time is 240min, and temperature is 45 ℃;
Described plant rennet is a bromelain; Described animal protease is pepsin and tryptic mixture.
Embodiment five
The preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 4 ︰ 1; The controlled condition of described catalyzing hydrolysis is 4 ︰ 1 for the enzyme dosage, and pH is 7, and the catalysis time is 240min, and temperature is 40 ℃;
Described plant rennet is the mixture of bromelain and papain; Described animal protease is pepsin and tryptic mixture.
Embodiment six
The preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 4 ︰ 1; The controlled condition of described catalyzing hydrolysis is 4 ︰ 1 for the enzyme dosage, and pH is 6, and the catalysis time is 240min, and temperature is 40 ℃;
Described plant rennet is a papain; Described animal protease is pepsin and tryptic mixture.
To embodiment one~five add up following table
Figure 2011101404441100002DEST_PATH_IMAGE002
The product that embodiment one~six is made detects, and gets following table
Figure 2011101404441100002DEST_PATH_IMAGE004

Claims (3)

1. the preparation method of marine alga active peptide may further comprise the steps:
1. with complex enzyme the food-grade sodium alginate is carried out catalyzing hydrolysis, get hydrolyzate, then this hydrolyzate is carried out inactivation, decolouring and filtration treatment, obtain marine alga polypeptide extract;
2. be that the NF membrane of 1000Da and 300Da is carried out purifying and concentrated with above-mentioned marine alga polypeptide extract by molecular weight, concentrate, then this concentrate is sterilized, after the sterilization, obtain marine alga active peptide powder by vacuum drying;
Step 1. in, described complex enzyme is that plant rennet and animal protease are with (the complex enzyme that the ratio of 2~4) ︰ 1 mixes; The controlled condition of described catalyzing hydrolysis is that (1~4) ︰ 1, pH are 5~7, and the catalysis time is 120~240min, and temperature is 40~60 ℃ for the enzyme dosage.
2. the preparation method of marine alga active peptide, it is characterized in that: described plant rennet is one or both in bromelain and the papain; Described animal protease is one or both in trypsase, the pepsin.
3. the preparation method of marine alga active peptide is characterized in that: step 1. in, the described complex enzyme complex enzyme that to be plant rennet and animal protease mix with the ratio of 4 ︰ 1; The controlled condition of described catalyzing hydrolysis is 4 ︰ 1 for the enzyme dosage, and pH is 6, and the catalysis time is 240min, and temperature is 40 ℃.
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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102961733A (en) * 2012-11-16 2013-03-13 福建省水产研究所 Application of seaweed polypeptide in preparing health-care product for reducing blood fat and blood sugar as well as beverage containing polypeptide for reducing blood fat and blood sugar
CN104432079A (en) * 2014-12-29 2015-03-25 陕西天宝大豆食品技术研究所 Whole-seaweed-peptide nourishment and preparation method thereof
CN107212422A (en) * 2017-06-23 2017-09-29 湖州知维技术服务有限公司 A kind of preparation method of algal active peptide
CN107254496A (en) * 2017-06-06 2017-10-17 威海温喜生物科技有限公司 A kind of preparation of seaweed amino acid and method of purification
CN108567141A (en) * 2018-05-04 2018-09-25 威海长青海洋科技股份有限公司 Kelp fish protein biology stimulin pulvis and its production method
CN111526721A (en) * 2017-12-18 2020-08-11 格玛实验室公司 Method for identifying and isolating bioactive compounds from seaweed extract
CN114538972A (en) * 2022-04-13 2022-05-27 广州世睿生物科技有限公司 Peptide-containing seaweed active substance and method for extracting seaweed active substance

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
何荣海等: "超声波在酶法生产紫菜降血压肽过程中的应用", 《江苏大学学报(自然科学版)》 *
王茵等: "海藻降血压肽的作用机制与制备", 《福建水产》 *
王茵等: "紫菜降血压肽酶法制备工艺的优化", 《福建水产》 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102961733A (en) * 2012-11-16 2013-03-13 福建省水产研究所 Application of seaweed polypeptide in preparing health-care product for reducing blood fat and blood sugar as well as beverage containing polypeptide for reducing blood fat and blood sugar
CN104432079A (en) * 2014-12-29 2015-03-25 陕西天宝大豆食品技术研究所 Whole-seaweed-peptide nourishment and preparation method thereof
CN107254496A (en) * 2017-06-06 2017-10-17 威海温喜生物科技有限公司 A kind of preparation of seaweed amino acid and method of purification
CN107212422A (en) * 2017-06-23 2017-09-29 湖州知维技术服务有限公司 A kind of preparation method of algal active peptide
CN111526721A (en) * 2017-12-18 2020-08-11 格玛实验室公司 Method for identifying and isolating bioactive compounds from seaweed extract
CN108567141A (en) * 2018-05-04 2018-09-25 威海长青海洋科技股份有限公司 Kelp fish protein biology stimulin pulvis and its production method
CN114538972A (en) * 2022-04-13 2022-05-27 广州世睿生物科技有限公司 Peptide-containing seaweed active substance and method for extracting seaweed active substance

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Denomination of invention: Preparation method of algal active peptide

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Pledgee: Nanxun Zhejiang rural commercial bank, Limited by Share Ltd and Fu sub branch

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