CN1896266B - Preparation of bioactive peptide for inhibiting meat fat oxidation - Google Patents
Preparation of bioactive peptide for inhibiting meat fat oxidation Download PDFInfo
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- CN1896266B CN1896266B CN200610036193A CN200610036193A CN1896266B CN 1896266 B CN1896266 B CN 1896266B CN 200610036193 A CN200610036193 A CN 200610036193A CN 200610036193 A CN200610036193 A CN 200610036193A CN 1896266 B CN1896266 B CN 1896266B
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Abstract
The present invention discloses a production method of biological active peptide that could inhibit the oxidation of fat in meat products with the following steps: the material is mixed with isopropyl alcohol of equal weight, then undergoes circular chemical extractions at 40-50DEG Cfor 1-2 hours; deionized water of equal weight is added and homogenized; the material is then added with protease and hydrolyzed at pH6.5-8.0, 50-65DEG C, and heated at 90DEG C for 20min to denature the protease when the degree of hydrolysis achieves 15-20%; the solid product is acquired through centrifugal filtration to isolate the supernatant, ultra-filtration, adsorption by macroporous resin, elution with 80% ethanol, collection of eluting peak, and vacuum concentration to eliminated ethanol. In this invention, the biological active peptide acquired through defatting, controlled enzymolysis, ultra-filtration and adsorption by macroporous resin can effectively inhibit the activity of the lipoxygenase, quench the free radical generated from lipid oxidation, and control the fat oxidation and outperformed standard of acid value in the storage of meat products.
Description
Technical field
The invention belongs to the biologically active peptides preparing technical field, relate in particular to a kind of preparation method with the biologically active peptides that suppresses meat fat oxidation.
Background technology
Multiple nutritional components such as meat product rich in proteins, fat, amino acid, VITAMIN are the requisites of human nutrition meals.Yet, the color burn that fats oxidn causes product takes place in the storage of meat product inevitably, the bad change of local flavor, downgrade, even lose edibleness.The reason of fats oxidn has photoxidation, autoxidation and enzymatic oxidn.Photoxidation is that oxidizing reaction generation hydroperoxide directly take place for unsaturated fatty acids and singlet oxygen.Autoxidation is the free chain reaction that grease takes place under the effect of light, heat, metal catalyst.Enzymatic oxidn is meant that the endogenous lipoxygenase catalytic oxygen and the grease reaction that exist in the meat product generate hydroperoxide.Free radical and hydroperoxide and degraded product thereof not only cause the bad change of meat product local flavor, and are one of inducements that causes modern diseases such as coronary heart disease, cancer.
At present, the method for oil-control oxidation mainly is to add antioxidant except that low-temperature dark storage, deoxidation packing.Yet synthetized oxidation preventive agent or effect are not good enough, or emulsifying property is poor, or security is under suspicion, and the overwhelming majority is used and is subjected to bigger restriction endogenous activity of fatty oxygenase unrestraint effect.Natural antioxidants (as herbaceous plant extract etc.) is not though there is food safety question, and use cost is higher and flavour of food products and color had bigger influence.In recent years, studies show that in a large number protein has certain anti-oxidant activity by the oligo peptide that the enzyme process control hydrolysis obtains, but have shortcomings such as consumption is big, effect is not good enough with the oil-control oxidation yet oligo peptide is directly used in the meat product.
Summary of the invention
The object of the present invention is to provide a kind of preparation method with the biologically active peptides that suppresses meat fat oxidation.Present method is by Virahol degreasing, control enzymolysis, ultrafiltration, macroporous resin adsorption, vacuum concentration, can obtain purity height, the oil-control oxidation effectiveness is remarkable, emulsifying property is good biologically active peptides.
For achieving the above object, the step of the present invention's employing is as follows:
Step 1: with after weight such as Virahol are mixed,, make feedstock fat content raw material less than 1% 40 ℃～50 ℃ following circulation lixiviates 1～2 hour;
Step 2: add the deionized water with weight such as raw materials, homogenate;
Step 3: add proteolytic enzyme, hydrolysis under ℃ condition of pH6.5～8.0,50～65, when degree of hydrolysis reached 15～20%, 90 ℃ of heating 20min inactivated proteases were with the molecular weight distribution of polypeptide in the control hydrolysis liquid;
Step 4: centrifuging obtains supernatant liquor, is the ultrafiltration membrance filter of 5000Da through molecular weight, gets through liquid;
Step 5: after adding salt and making salts contg through liquid be 0.2mol/L～0.4mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, must solid phase prod.
Raw material in step 1 and the step 2 is meant in pork or chicken or the sardines meat any one.
The proteolytic enzyme that adds in the step 3 is business-like food-grade albumen enzyme: Protamex
TMOr Alcalase
TM
In the step 3 in the hydrolyzed solution molecular weight distribution of polypeptide concentrate on 1000Da～5000Da.
Content of peptides accounts for 65%～80% of total protein content in the step 5 gained solid phase prod.
The present invention adopts the Virahol degreasing, has controlled fatty content, reduces fat and has the detrimentally affect that causes; By control hydrolysis polypeptide molecular weight is controlled between 1000Da～5000Da; By ultrafiltration and macroporous resin adsorption enrichment, purifying biological bioactive peptide.
The present invention compared with prior art has following advantage and beneficial effect:
(1) the present invention adopts the Virahol degreasing, has controlled fatty content, and grease reduces its influence to the anti-oxidation characteristics of enzymolysis solution in very easily oxidation of enzymolysis process;
(2) the present invention obtains good, the active high biologically active peptides of emulsifying property by ultrafiltration, macroporous resin adsorption;
(3) biologically active peptides of the present invention preparation is safe, and addition is little can not exert an adverse impact to the meat product local flavor.
Description of drawings
Fig. 1 is for adding 0.1% (accounting for the percentage composition of sausage total mass) with the biologically active peptides (is feedstock production with pork) of present method preparation synoptic diagram that influences to acid value in the Guangdong style sausage storage;
Fig. 2 is for adding 0.1% (accounting for the percentage composition of sausage total mass) with the biologically active peptides (is feedstock production with pork) of present method preparation synoptic diagram that influences to peroxide value in the Guangdong style sausage storage;
Fig. 3 is for adding with the biologically active peptides (is feedstock production with pork) of present method preparation clearance rate synoptic diagram to free radical DPPH;
Fig. 4 influences synoptic diagram for the biologically active peptides (is feedstock production with pork) that adds with present method preparation to activity of fatty oxygenase.
Embodiment
For a better understanding of the present invention, the invention will be further described below in conjunction with embodiment and accompanying drawing.
Pork is mixed with weight such as Virahols, and at 50 ℃ of following circulation lixiviate 1hr, lipid content is 0.9% in the pork; Add the deionized water with weight such as porks, homogenate; Add Protamex
TM2000U/100g meat gruel liquid at pH6.5, carries out enzyme digestion reaction under 50 ℃, and degree of hydrolysis reaches 20% back 90 ℃ of heating 20min inactivated proteases, and supernatant liquor is got in centrifuging, and adopting molecular weight is to obtain seeing through liquid after the ultra-filtration membrane ultrafiltration of 5000Da; After adding salt and making salts contg through liquid be 0.2mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb to seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, get solid phase prod, content of peptides accounts for 77% of total protein content in the gained solid phase prod.
Pork is mixed with weight such as Virahols, and at 40 ℃ of following circulation lixiviate 1hr, lipid content is 0.9% in the pork; Add the deionized water with weight such as porks, homogenate; Add Alcalase
TM2000U/100g meat gruel liquid at pH6.5, carries out enzyme digestion reaction under 50 ℃, and degree of hydrolysis reaches 19% back 90 ℃ of heating 20min inactivated proteases, and supernatant liquor is got in centrifuging, and adopting molecular weight is to obtain seeing through liquid after the ultra-filtration membrane ultrafiltration of 5000Da; After adding salt and making salts contg through liquid be 0.25mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb to seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, get solid phase prod, content of peptides accounts for 78% of total protein content in the gained solid phase prod.
Sardines meat is mixed with weight such as Virahols, and at 45 ℃ of following circulation lixiviate 1.5hr, lipid content is 0.7% in the sardines meat; Add the deionized water with weight such as sardines meat, homogenate; Add Protamex
TMThe 2000U/100g flesh of fish, at pH6.5, carry out enzyme digestion reaction under 50 ℃, degree of hydrolysis reaches 16% back 90 ℃ of heating 20min inactivated proteases, supernatant liquor is got in centrifuging, adopting molecular weight is to obtain seeing through liquid after the ultra-filtration membrane ultrafiltration of 5000Da, after adding salt and making salts contg through liquid be 0.3mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb to seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, gets solid phase prod, and content of peptides accounts for 67% of total protein content in the gained solid phase prod.
Sardines meat is mixed with weight such as Virahols, and at 47 ℃ of following circulation lixiviate 1.5hr, lipid content is 0.7% in the sardines meat; Add the deionized water with weight such as sardines meat, homogenate; Add Alcalase
TMThe 2000U/100g flesh of fish, at pH6.0, carry out enzyme digestion reaction under 55 ℃, degree of hydrolysis reaches 17% back 90 ℃ of heating 20min inactivated proteases, supernatant liquor is got in centrifuging, adopting molecular weight is to obtain seeing through liquid after the ultra-filtration membrane ultrafiltration of 5000Da, after adding salt and making salts contg through liquid be 0.35mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb to seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, gets solid phase prod, and content of peptides accounts for 66% of total protein content in the gained solid phase prod.
Embodiment 5
Chicken is mixed with weight such as Virahols, and at 48 ℃ of following circulation lixiviate 1.5hr, making lipid content is 0.46%; Add the deionized water with weight such as chicken, homogenate; Add Alcalase2500U/100g meat gruel liquid, at pH7.5, carry out enzyme digestion reaction under 60 ℃, degree of hydrolysis reaches 15% back 90 ℃ of heating 20min inactivated proteases; Supernatant liquor is got in centrifuging, adopting molecular weight is to obtain seeing through liquid after the ultra-filtration membrane ultrafiltration of 5000Da, after adding salt and making salts contg through liquid be 0.4mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb to seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, gets solid phase prod, and content of peptides accounts for 71% of total protein content in the gained solid phase prod.
Chicken is mixed with weight such as Virahols, and at 44 ℃ of circulation lixiviate 2hr, making lipid content is 0.46%; Add the deionized water with weight such as chicken, homogenate; Add Alcalase2500U/100g meat gruel liquid, at pH8.0, carry out enzyme digestion reaction under 65 ℃, degree of hydrolysis reaches 15% back 90 ℃ of heating 20min inactivated proteases; Supernatant liquor is got in centrifuging, adopting molecular weight is to obtain seeing through liquid after the ultra-filtration membrane ultrafiltration of 5000Da, after adding salt and making salts contg through liquid be 0.4mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb to seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, gets solid phase prod, and content of peptides accounts for 72% of total protein content in the gained solid phase prod.
As seen from Figure 1, the biological activity Toplink of adding the preparation of 0.1% present method significantly suppresses the acid value of Guangdong style sausage in storage process and raises, the acid value of room temperature storage Guangdong style sausage after 80 days reaches 3.25mg/g, and the acid value that contrasts (not adding biologically active peptides) is up to 7.12mg/g.
As seen from Figure 2, the biological activity Toplink of adding the preparation of 0.1% present method significantly suppresses the peroxide value of Guangdong style sausage in storage process and raises, the peroxide value of room temperature storage Guangdong style sausage after 80 days reaches 0.45mg/100g, and the peroxide value that contrasts (not adding biologically active peptides) is up to 0.78mg/100g.
As seen from Figure 3, the biologically active peptides of present method preparation has the activity of removing free radical DPPH, its IC under lower concentration
50Be 0.145mg/ml.
As seen from Figure 4, the biologically active peptides of present method preparation has the activity that suppresses lipoxygenase, its IC under lower concentration
50Be 2.6mg/ml.
Claims (5)
1. preparation method with the biologically active peptides that suppresses meat fat oxidation is characterized in that preparation process is as follows:
Step 1: with after weight such as Virahol are mixed,, make feedstock fat content raw material less than 1% 40 ℃～50 ℃ following circulation lixiviates 1～2 hour;
Step 2: add the deionized water with weight such as raw materials, homogenate;
Step 3: add proteolytic enzyme, hydrolysis under ℃ condition of pH6.5～8.0,50～65, when degree of hydrolysis reached 15～20%, 90 ℃ of heating 20min inactivated proteases were with the molecular weight distribution of polypeptide in the control hydrolysis liquid;
Step 4: centrifuging obtains supernatant liquor, is the ultrafiltration membrance filter of 5000Da through molecular weight, gets through liquid;
Step 5: after adding salt and making salts contg through liquid be 0.2mol/L～0.4mol/L, adopt DA201-C type macroporous adsorbent resin to adsorb seeing through liquid, collect elution peak with 80% ethanol as strippant, vacuum concentration boils off ethanol, must solid phase prod.
2. the preparation method with the biologically active peptides that suppresses meat fat oxidation according to claim 1 is characterized in that raw material in described step 1 and the step 2 is meant in pork or chicken or the sardines meat any one.
3. the preparation method with the biologically active peptides that suppresses meat fat oxidation according to claim 1 is characterized in that the proteolytic enzyme that adds in the described step 3 is business-like food-grade albumen enzyme: Protamex
TMOr Alcalase
TM
4. the preparation method with the biologically active peptides that suppresses meat fat oxidation according to claim 1 is characterized in that in the described step 3 that the molecular weight distribution of polypeptide concentrates on 1000Da～5000Da in the hydrolyzed solution.
5. the preparation method with the biologically active peptides that suppresses meat fat oxidation according to claim 1 is characterized in that content of peptides accounts for 65%～80% of total protein content in the described step 5 gained solid phase prod.
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| CN200610036193A CN1896266B (en) | 2006-06-30 | 2006-06-30 | Preparation of bioactive peptide for inhibiting meat fat oxidation |
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| CN200610036193A CN1896266B (en) | 2006-06-30 | 2006-06-30 | Preparation of bioactive peptide for inhibiting meat fat oxidation |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104872371A (en) * | 2015-06-08 | 2015-09-02 | 宁波希诺亚海洋生物科技有限公司 | Preparation method for small peptide protein of sardine |
| CN112521477B (en) * | 2021-01-20 | 2022-07-08 | 福建农林大学 | Preparation method and application of antioxidant color fixative for natural meat products |
| CN112931759A (en) * | 2021-03-13 | 2021-06-11 | 绵阳市米小福食品有限公司 | Method for removing peculiar smell of raw meat |
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Non-Patent Citations (5)
| Title |
|---|
| Hui-Chun Wu et al.Free amino acids and peptides as related to antioxidantproperties in proteinh hydrolysates of mackerel (Scomberaustriasicus).Food Research International36.2003,36949-957. * |
| 李琳等.利用人工神经网络优化制备鳙鱼抗氧化肽.四川大学学报（工程科学版）38 1.2006,38(1),第80-85页. |
| 李琳等.利用人工神经网络优化制备鳙鱼抗氧化肽.四川大学学报(工程科学版)38 1.2006,38(1),第80-85页. * |
| 李琳等.鳙鱼蛋白酶解液清除自由基的研究.水产科学24 10.2005,24(10),15-18. |
| 李琳等.鳙鱼蛋白酶解液清除自由基的研究.水产科学24 10.2005,24(10),15-18. * |
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Owner name: DONGGUAN XUJI FOOD CO., LTD. Free format text: FORMER OWNER: SOUTHERN CHINA UNIVERSITY OF TECHNOLOGY Effective date: 20101223 Free format text: FORMER OWNER: GUANGDONG PROV. FOOD INDUSTRY INST. |
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Free format text: CORRECT: ADDRESS; FROM: 510640 NO.381, WUSHAN ROAD, TIANHE DISTRICT, GUANGZHOU CITY, GUANGDONG PROVINCE TO: 523110 ZHOUWU INDUSTRIAL AREA, DONGCHENG DISTRICT, DONGGUAN CITY, GUANGDONG PROVINCE |
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Effective date of registration: 20101223 Address after: 523110, Dongcheng District, Guangdong, Dongguan weeks Housing Industrial Zone Patentee after: Dongguan Xuji Food Co., Ltd. Address before: 510640 Tianhe District, Guangdong, No. five road, No. 381, Co-patentee before: Guangdong Prov. Food Industry Inst. Patentee before: South China University of Technology |
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Granted publication date: 20100526 Termination date: 20190630 |