CN101580861B - Biologically active peptide capable of inhibiting after-fermentation of acidified milk and preparation method thereof - Google Patents
Biologically active peptide capable of inhibiting after-fermentation of acidified milk and preparation method thereof Download PDFInfo
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- CN101580861B CN101580861B CN2009100400465A CN200910040046A CN101580861B CN 101580861 B CN101580861 B CN 101580861B CN 2009100400465 A CN2009100400465 A CN 2009100400465A CN 200910040046 A CN200910040046 A CN 200910040046A CN 101580861 B CN101580861 B CN 101580861B
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Abstract
The invention provides a biologically active peptide capable of inhibiting after-fermentation of acidified milk and a preparation method thereof. The method comprises the following steps: performing maltodextrin graft modification on lactoprotein so that the grafting degree of the lactoprotein reaches between 4 and 10 percent; adding proteinase to perform zymohydrolysis at a temperature of between45 and 60 DEG C, wherein when the proteinase is neutral proteinase or alkali protease, the pH during the zymohydrolysis is between 5.0 and 8.0, and when the proteinase is acid proteinase, the pH duri ng the zymohydrolysis is between 2.0 and 4.0; when the degree of hydrolysis reaches between 5 and 10 percent, heating the mixture at a temperature of between 85 and 95 DEG C for 15 to 30 minutes to inactivate enzymes and end the zymohydrolysis so as to control the molecular weight of the biologically active peptide; and processing enzymatic hydrolyzate with inactivated enzymes through a macroporous resin to collect the unadsorbed component so as to obtain the biologically active peptide. The method can obtain the biologically active peptide which can remarkably inhibit the after-fermentation of the acidified milk through the maltodextrin graft modification, controllable zymohydrolysis and macroporous resin enrichment.
Description
Technical field
The present invention relates to the technical field that suppresses the fermented-milk secondary fermentation, is specifically related to have biologically active peptides that suppresses the fermented-milk secondary fermentation and preparation method thereof..
Background technology
Milk-product are the most approaching perfect food, almost include biological required whole nutritive ingredients, as materials such as energy, nutrition, physiological regulation.Fermented-milk is the product that utilizes milk-acid bacteria (as lactobacillus bulgaricus and thermophilus streptococcus) that cow's milk is fermented and forms.During the fermentation, milk-acid bacteria can produce a large amount of lactic acid, beneficial source of students and flavour substances, and these products all play important effect to adjusting intestinal microflora balance, prevention courses of infection and digestive tract diseases directly or indirectly.In addition, milk-acid bacteria can also be synthesized some special enzyme system and cell surface materials, can play reducing cholesterol, stops pathogenic bacterium that enteron aisle is invaded, and increases immunologic function and anticancer effect.Just be based on above reason, the fermented-milk industry development is rapid, has become indispensable staple product in China's people's lives consumption.
Thermophilus streptococcus and lactobacillus bulgaricus have growth complementary effect or are called symbiosis (Mutualism) in the fermented-milk fermenting process, and the growth of coccus can promote the growth of bacillus simultaneously, and the meta-bolites that the bacillus growth forms guarantees the growth of coccus.Bacterium lacticum is decomposing protein, produces acid the contribution of fermented-milk, and suis has vital role to the excellent flavor and the organoleptic quality of fermented-milk.Yet fermented-milk is fermenting to certain acidity, need be cooled between 0～5 ℃, to reduce milk-acid bacteria metabolism ability, produces acid to prevent the Bacterium lacticum excessing metabolism, causes the overweight and generation bitter taste of tart flavour.Even under 0～5 ℃ of such refrigerated condition, the back acidifying phenomenon of fermented-milk is still very serious, the value preserving phase that this has shortened product greatly, has a strong impact on the local flavor organoleptic quality of fermented milk prod.The back acidifying of fermented-milk mainly is that lactobacillus bulgaricus continues to produce sour causing in the preservation process.
Extensively the lactobacillus bulgaricus that adopts is separated from milk the earliest, and milk is its natural medium.It is generally acknowledged that it is a kind of milk source property milk-acid bacteria, the secondary fermentation of fermented-milk mainly is that lactobacillus bulgaricus continues to produce sour causing under acidic conditions.Therefore, have the secondary fermentation of control fermented-milk, prolong the fermented-milk quality guaranteed period,, have great economic worth and realistic meaning the fermented-milk industry by in fermented-milk, adding biologically active peptides with inhibition secondary fermentation.
Summary of the invention
The present invention seeks to overcome the prior art above shortcomings, provide to have biologically active peptides that suppresses the fermented-milk secondary fermentation and preparation method thereof.For achieving the above object, the present invention is by the following technical solutions:
Have the preparation method of the biologically active peptides that suppresses the fermented-milk secondary fermentation, comprise the steps:
(1) milk protein solution is carried out the maltodextrin graft modification, make the graft(ing) degree of milk-protein reach 4～10%;
(2) add proteolytic enzyme and carry out enzymolysis under 45 ℃～60 ℃ condition, when described proteolytic enzyme was neutral protease or Sumizyme MP, pH was 5.0～8.0 during enzymolysis, and when described proteolytic enzyme is aspartic protease, pH is 2.0～4.0 during enzymolysis;
(3) when degree of hydrolysis reaches 5%～10%, 85～95 ℃ of heating 15～30min go out the enzyme enzymolysis reaction to control the molecular weight of biologically active peptides;
(4) will go out enzymolysis solution that enzyme lives by macroporous resin treatment, collect not absorbed component, promptly get described biologically active peptides.Among the above-mentioned preparation method, described milk-protein is Sodium Caseinate, whey-protein or casein.
Among the above-mentioned preparation method, contain 4g/100ml～8g/100ml milk-protein in the described graft modification in the milk protein solution, 2g/100ml～3g/100ml maltodextrin, ultrasonic grafting is handled.
Among the above-mentioned preparation method, described proteolytic enzyme is business-like food-grade albumen enzyme, and this proteolytic enzyme is papoid, Sumizyme MP, stomach en-, Quimotrase or trypsinase.
Among the above-mentioned preparation method, described macroporous resin is XAD16 or XAD1180 type.
Among the above-mentioned preparation method, the molecular weight of biologically active peptides is 500～5000Da, and peptide content accounts for 60～80% (w/w) of total protein content.
Among the above-mentioned preparation method, the protein recovery of milk-protein is 70～85%.
Among the above-mentioned preparation method, the condition that ultrasonic grafting is handled is: 70～80 ℃, and pH6.5～7.5, ultrasonic frequency is 15kHZ, the treatment time is 15～30min.
The present invention has adopted above technical scheme, has following advantage and effect:
1, gained biological activity peptide molecular weight of the present invention concentrates between 500～5000Da, and side-chain radical contains glycosyl, has the effect of significant inhibition fermented-milk secondary fermentation.
2, the casein hydrophobicity is strong, and the enzymolysis product bitter taste is outstanding.The present invention is by carrying out glycosylation modification to the casein side-chain radical, shielding bitter taste group, change caseic restriction enzyme site, significantly reduced the content of hydrophobicity bitter peptides in the biologically active peptides, and dispel minority bitter peptides in the enzymolysis solution by macroporous resin adsorption, so the gained biologically active peptides does not have bitter taste.
Description of drawings
Fig. 1 produces the influence curve of acid and does not add the pairing influence curve of biologically active peptides the fermented-milk shelf lives for adding 500ppm (w/w) biologically active peptides in the embodiment.
Fig. 2 for the biologically active peptides that adds 500ppm (w/w) in the embodiment to the active influence curve of fermented-milk shelf lives coccus with do not add the pairing influence curve of biologically active peptides.
Specific embodiments
Below in conjunction with drawings and Examples concrete enforcement of the present invention is described further.
Embodiment 1: utilize casein production to have the biologically active peptides that acid is produced in significant inhibition fermented-milk secondary fermentation
1, the graft modification of material protein: the casein consumption is that 4g/100ml and maltodextrin consumption are 2g/100ml in the milk protein solution, 70 ℃, and pH6.5, ultrasonic frequency is 15kHZ, the supersound process time is that to make the casein graft(ing) degree be 4.1% to 15min..
2, Controlled-enzymatic Hydrolysis: adopt sodium hydroxide to transfer milk protein solution pH to pH8.0, add Sumizyme MP Alcalase, 60 ℃ are carried out enzyme digestion reaction, adopt pH-Stat method control hydrolysis degree to 10%, get enzymolysis solution.
3, the enzymolysis solution enzyme that goes out: 85 ℃ of heating 30min enzyme that goes out, enzymolysis reaction.
4, remove by filter insoluble substance.
5, enzymolysis solution is crossed the XAD16 macroporous resin, collected not absorbed component, promptly obtain having the biologically active peptides that acid is produced in significant inhibition fermented-milk secondary fermentation.The protein recovery of raw material reaches 72%, and the biological activity peptide molecular weight mainly concentrates between 500～5000, and peptide content accounts for 68% of total protein content.
Add 500ppm (w/w) embodiment 1 gained biologically active peptides Fig. 1 and Fig. 2 are seen in the influence of fermented-milk shelf lives product acid.As seen from Figure 1, producing sour measuring after interpolation biologically active peptides secondary fermentation breast stores 30 days only is 30% of blank sample, and this shows that the biologically active peptides of 500ppm can obviously suppress to produce acid in the fermented-milk storage process, keeps the fermented milk prod local flavor, extends the shelf life.In addition, behind the interpolation biologically active peptides, coccus quantity also than the high order of magnitude of blank group, is seen Fig. 2 in the fermented milk prod.This shows that this biologically active peptides can prevent fermented milk prod coccus death in storage process, keeps product quality.
Embodiment 2: utilize casein production to have the biologically active peptides that acid is produced in significant inhibition fermented-milk secondary fermentation
1, the graft modification of material protein: the Sodium Caseinate consumption is that 6g/100ml and maltodextrin consumption are 3g/100ml in the milk protein solution, 75 ℃, and pH7.0, ultrasonic frequency is 15kHZ, ultrasonic time is that to make the casein graft(ing) degree be 6.1% to 20min..
2, Controlled-enzymatic Hydrolysis: adopt hydrochloric acid to transfer milk protein solution pH to 3.0, add stomach en-, 52 ℃ are carried out enzyme digestion reaction, adopt pH-Stat method control hydrolysis degree to 5.1%, get enzymolysis solution.
3, the enzymolysis solution enzyme that goes out: 85 ℃ of heating 30min enzyme that goes out, enzymolysis reaction.
4, remove by filter insoluble substance.
5, enzymolysis solution is crossed the XAD16 macroporous resin, collected not absorbed component, promptly obtain having the biologically active peptides that acid is produced in significant inhibition fermented-milk secondary fermentation.The protein recovery of raw material reaches 78%, and mainly between 500～5000Da, peptide content accounts for 76% of total protein content to the biological activity peptide molecular weight.
Embodiment 3: utilize whey-protein production to have the biologically active peptides that acid is produced in significant inhibition fermented-milk secondary fermentation
1, the graft modification of material protein: the whey-protein consumption is that 8g/100ml and maltodextrin consumption are 2.5g/100ml in the milk protein solution, 80 ℃, and pH7.5, ultrasonic frequency is 15kHZ, ultrasonic time is that to make the casein graft(ing) degree be 9.2% to 30min..
2, Controlled-enzymatic Hydrolysis: adopt hydrochloric acid to transfer milk protein solution pH to 7.0, add papoid, 55 ℃ are carried out enzyme digestion reaction, adopt pH-Stat method control hydrolysis degree to 8%, get enzymolysis solution.
3, the enzymolysis solution enzyme that goes out: 95 ℃ of heating 20min enzyme that goes out, enzymolysis reaction.
4, remove by filter insoluble substance.
5, enzymolysis solution is crossed the XAD1180 macroporous resin, collected not absorbed component, promptly obtain having the biologically active peptides that acid is produced in significant inhibition fermented-milk secondary fermentation.The protein recovery of raw material reaches 80.2%, and the biological activity peptide molecular weight mainly concentrates between 500～5000, and peptide content accounts for 79% of total protein content.
Claims (4)
1. have the preparation method of the biologically active peptides that suppresses the fermented-milk secondary fermentation, it is characterized in that comprising the steps:
(1) milk protein solution is carried out the maltodextrin graft modification, make the graft(ing) degree of milk-protein reach 4%～10%; Described milk-protein is a casein; In the described graft modification in the milk protein solution milk protein content be 4g/100ml～8g/100ml, the content of maltodextrin is 2g/100ml～3g/100ml, adopts ultrasonic grafting to handle; The condition that ultrasonic grafting is handled is: 70～80 ℃, and pH6.5～7.5, ultrasonic frequency is 15kHZ, the treatment time is 15～30min;
(2) add proteolytic enzyme under 45 ℃～60 ℃ condition and carry out enzymolysis, get enzymolysis solution, when described proteolytic enzyme was neutral protease or Sumizyme MP, pH was 5.0～8.0 during enzymolysis; When described proteolytic enzyme was aspartic protease, pH was 2.0～4.0 during enzymolysis; Described proteolytic enzyme is the food-grade albumen enzyme, and described neutral protease is papoid, Quimotrase or trypsinase, and described Sumizyme MP is Alcalase, and described aspartic protease is a stomach en-;
(3) heating when degree of hydrolysis reaches 5%～10%, enzyme 15～30min goes out under 85～95 ℃; Enzymolysis reaction is with the molecular weight of control biologically active peptides;
(4) will go out enzymolysis solution that enzyme lives by macroporous resin treatment, and collect not absorbed component, and promptly get described biologically active peptides, the molecular weight of biologically active peptides is 500～5000Da, and peptide content accounts for 60～80% (w/w) of total protein content; Described macroporous resin is XAD16 or XAD1180 type.
2. method according to claim 1, the protein recovery that it is characterized in that milk-protein is 70%～85%.
3. method according to claim 2 is characterized in that also enzymolysis solution being removed by filter insoluble substance after step (3) enzymolysis reaction.
4. has a biologically active peptides that suppresses the fermented-milk secondary fermentation by what the described preparation method of claim 1 made.
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| CN101993476B (en) * | 2010-09-10 | 2013-06-12 | 华南理工大学 | Casein active single peptide as well as preparation method and application thereof |
| CN103392907B (en) * | 2013-06-08 | 2014-12-17 | 东北农业大学 | Method for improvement of whey protein nano-fiber polymerization amount by enzymatic modification |
| CN108467487B (en) * | 2018-03-30 | 2020-09-04 | 合肥工业大学 | Starch-based dextrin modified casein polypeptide conjugate, preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1135266C (en) * | 2000-03-01 | 2004-01-21 | 刘树军 | Process and equipment for preparing low-molecular peptide from sodium caseinate as raw material |
| CN101176486A (en) * | 2007-12-04 | 2008-05-14 | 内蒙古蒙牛乳业（集团）股份有限公司 | Sterilizing flavouring sour milk and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1135266C (en) * | 2000-03-01 | 2004-01-21 | 刘树军 | Process and equipment for preparing low-molecular peptide from sodium caseinate as raw material |
| CN101176486A (en) * | 2007-12-04 | 2008-05-14 | 内蒙古蒙牛乳业（集团）股份有限公司 | Sterilizing flavouring sour milk and preparation method thereof |
Non-Patent Citations (1)
| Title |
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| 秦宜德.乳蛋白的主要组分及其研究现状.《生物学杂志》.2003,第20 卷(第2 期),5-7. * |
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Application publication date: 20091118 Assignee: Zhaoqing Huanfa Biotechnology Co., Ltd. Assignor: South China University of Technology Contract record no.: 2013440000144 Denomination of invention: Biologically active peptide capable of inhibiting after-fermentation of acidified milk and preparation method thereof Granted publication date: 20111228 License type: Exclusive License Record date: 20130418 |
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