CN107365823A - A kind of preparation method of freshwater mussel antioxidation active peptides - Google Patents
A kind of preparation method of freshwater mussel antioxidation active peptides Download PDFInfo
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Abstract
The present invention relates to a kind of preparation method of freshwater mussel antioxidation active peptides, belong to food processing technology field.The present invention contains abundant polysaccharide using freshwater mussel meat, unrighted acid and mineral element, it is a kind of high-quality Edible resource, and aboundresources, yield is big, protein content is high, it is to prepare the high-quality raw material sources of antioxidation active peptides, and price is very cheap, steady sources, microbial fermentation is made full use of with digesting respective advantage, make protein degradation more abundant, small peptide degree is higher, pass through Maillard reaction simultaneously, assign food special color and luster and smell, avoid the bitter taste of enzymolysis product, improve product palatability, improve polypeptide products quality and nutritive value, make it have anti-mutagenesis, the characteristic such as antiviral, further enhance antioxidation activity.
Description
Technical field
The present invention relates to a kind of preparation method of freshwater mussel antioxidation active peptides, belong to food processing technology field.
Background technology
Reactive oxygen species and free radicals are resulted from normal physiological activity, and the cell of the especially mankind and other aerobionts are exhaled
Suction acts on.Research finds that active oxygen and free radical are easily caused many diseases, as atherosclerosis, hypertension, inflammation, cancer,
Diabetes, Alzheimer disease etc..Therefore, people get over to the relation caused by body oxidation between free radical and aging and disease
More to pay attention to.Antioxidant is also widely used in the industries such as food, health products, cosmetics, medicine therewith, but uses at present
Major part be artificial synthetized oxidation preventive agent, many potential safety hazards be present, it is long-term use of even can be in machine cylinder accumulation toxin
And there is carcinogenic danger etc..In recent years, the peptide matters between amino acid and protein gradually draw attention, and research finds that it has
Have more preferable antioxidation activity, Stability Analysis of Structures, molecular weight it is small, it is easy absorb, without adverse immune react, be excellent antioxidant,
Compared with other antioxidants, there is more wide development and application prospect.
With the utilization of aquatic biological resource, the research that biologically active peptide is prepared on aquatic product protein is also gradually opened up
Open.Under normal circumstances, active peptide is made up of 3~20 amino acid, and molecular weight is generally less than 6ku, and many research reports show biology
Active peptide has multiple functions characteristic, such as hypotensive, anti-oxidant, anti-inflammatory and antibacterial.Numerous studies show by aquatic product protein system
Standby anti-oxidation peptide has inhibitory action for different oxidation systems.
Wild freshwater mussel is widely distributed, cheap, and 2013-2014 is only 0.7 yuan/kg.Freshwater mussel is precious from head to foot, is had rich
Rich nutritive value, protein content (being all contents on dry basis below) is up to 51.07% in mussel meat, and amino acid classes are complete, must
Amino acid is needed to account for the 44.43% of total amino acid content, higher than the recommendation (35.38%) of WHO/FAO patterns.In addition, mussel meat also contains
There are abundant polysaccharide, unrighted acid and mineral element, be a kind of low price, high-quality available resources, but because its meat is slightly tough,
Processing and utilization degree is relatively low, and resource utilization is extremely low, wastes seriously, and the method for preparing anti-oxidation peptide at present is mainly traditional enzyme
Solution, its reaction condition is gentle, and beneficial to control, maximizing obtains target peptide fragment.But individually prepared using enzymatic isolation method anti-oxidant
During peptide, small scale is digested, the target peptide fragment of acquisition is few, and commercial enzyme cost is high.
Therefore, it is right for research of the freshwater mussel anti-oxidation peptide as functional food, medicine and cosmetic and function factor
Widen freshwater mussel intensive processing approach, realize wild freshwater mussel higher value application, have important practical significance and good economy is imitated
Benefit.
The content of the invention
The technical problems to be solved by the invention：It is cheap for the wild freshwater mussel aboundresources in China, but because of its meat
Thick tough, processing and utilization degree is relatively low, and resource utilization is extremely low, wastes serious；The method for preparing anti-oxidation peptide at present mainly passes
System enzymatic isolation method, its reaction condition is gentle, and beneficial to control, maximizing obtains target peptide fragment.But individually prepared using enzymatic isolation method anti-
When aoxidizing peptide, small scale, the problems such as target peptide fragment of acquisition is few, and commercial enzyme cost is high are digested, there is provided a kind of freshwater mussel is anti-oxidant
The preparation method of active peptide.
In order to solve the above technical problems, the technical solution adopted by the present invention is：
A kind of preparation method of freshwater mussel antioxidation active peptides, it is characterised in that specifically preparation process is：
（1）Take fresh freshwater mussel to shell, remove internal organ after wash, and be transferred to meat grinder rub after cross colloid mill, add deionized water
Solid content is adjusted, obtains mussel meat homogenate；
（2）Mussel meat homogenate, urea, potassium hydrogen phosphate, sodium chloride are taken, simultaneously autoclave sterilization is well mixed, obtains fermentation substrate；
（3）By inoculum concentration 2.5~5.0%（v/v）Bacillus subtilis is seeded in fermentation substrate and fermented, after fermentation ends from
Heart separation takes supernatant, obtains freshwater mussel zymotic fluid；
（4）Deionized water regulation substrate protein mass fraction and regulation system pH are added into freshwater mussel zymotic fluid, adds alkalescence
Protease hydrolyzed, centrifuged after sterilizing and take supernatant liquor, be freeze-dried to obtain freshwater mussel Gly-His-Lys；
（5）Reduced sugar and freshwater mussel Gly-His-Lys mixing addition are gone from water, regulation system pH, and heating response, after completion of the reaction from
The heart separates, and takes upper liquid to be evaporated to dryness, obtains freshwater mussel antioxidation active peptides.
Step（1）The solid content is 5~10%.
Step（2）The mussel meat homogenate, urea, potassium hydrogen phosphate, the parts by weight of sodium chloride are 1000~2000 portions of freshwater mussels
Meat is homogenized, 15~30 parts of urea, 3~6 parts of potassium hydrogen phosphates, 5~10 parts of sodium chloride.
Step（3）The fermentation process is oscillation and fermentation 2~3 days at 32~38 DEG C.
Step（4）The substrate protein mass fraction is 5~6%, and regulation system pH is 9.5~10.0, by enzyme bottom ratio
200U/g adds alkali protease, and 3~4h is digested at 50~60 DEG C.
Step（5）The reduced sugar is the one or more in glucose, lactose, maltose, galactolipin, xylose.
Step（5）The regulation system pH is 11~12, is heated to 90~100 DEG C of 3~4h of reaction.
Compared with other method, advantageous effects are the present invention：
（1）Freshwater mussel meat contains abundant polysaccharide, unrighted acid and mineral element, is a kind of high-quality Edible resource, and resource
Abundant, yield is big, and protein content is high, is to prepare the high-quality raw material sources of antioxidation active peptides, and price is very cheap, source
It is stable；
（2）The present invention makes full use of microbial fermentation to make protein degradation more abundant, small peptide degree is more with digesting respective advantage
Height, while by Maillard reaction, assign food special color and luster and smell, avoid the bitter taste of enzymolysis product, improve production
Product palatability, polypeptide products quality and nutritive value are improved, make it have the characteristic such as anti-mutagenesis, antiviral, further enhance
Antioxidation activity.
Embodiment
Take fresh freshwater mussel to shell, remove internal organ after wash 2~3 times, and be transferred to meat grinder rub after cross colloid mill, add
Ionized water, it is 5~10% to adjust to solid content, obtains mussel meat homogenate, takes 1~2kg mussel meats to be homogenized, 15~30g urea, 3~6g
Potassium hydrogen phosphate, 5~10g sodium chloride, 20~30min is stirred with 300~400r/min, and be transferred in autoclave, 121~125
15~20min of autoclaving at DEG C, obtains fermentation substrate, by inoculum concentration 2.5~5.0%（v/v）Bacillus subtilis is seeded to hair
In ferment matrix, oscillation and fermentation 2~3 days, are transferred in centrifuge after fermentation ends at 32~38 DEG C, with 6000~8000r/min
15~20min is centrifuged, supernatant is taken, obtains freshwater mussel zymotic fluid, deionized water regulation substrate protein is added into freshwater mussel zymotic fluid
White mass fraction is 5~6%, and regulation system pH is 9.5~10.0, then adds alkali protease than 200U/g by enzyme bottom, 50
3~4h is digested at~60 DEG C, then is transferred in autoclave, 15~20min of autoclaving at 121~125 DEG C, after being cooled to room temperature
It is fitted into centrifuge, 15~20min is centrifuged with 6000~8000r/min, takes supernatant liquor to be placed in freeze drying box,
15~20h is freeze-dried at -20~-15 DEG C, obtains freshwater mussel Gly-His-Lys, 30~60g reduced sugars is taken, 10~20g freshwater mussel Gly-His-Lys, adds
In 400~800mL deionized waters, 20~30min is stirred with 300~400r/min, regulation system pH is 11~12, and is heated to
90~100 DEG C, it is fitted into after reacting 3~4h in centrifuge, 15~20min is centrifuged with 6000~8000r/min, takes upper liquid
It is placed in Rotary Evaporators, is evaporated to dryness at 50~60 DEG C, obtains freshwater mussel antioxidation active peptides.
Example 1
Take fresh freshwater mussel to shell, remove internal organ after wash 2 times, and be transferred to meat grinder rub after cross colloid mill, add deionized water,
Regulation to solid content is 5%, obtains mussel meat homogenate, takes 1kg mussel meats to be homogenized, 15g urea, 3g potassium hydrogen phosphates, 5g sodium chloride, with
300r/min stirs 20min, and is transferred in autoclave, and 15~20min of autoclaving, obtains fermentation substrate at 121 DEG C, by inoculation
Amount 2.5%（v/v）Bacillus subtilis is seeded in fermentation substrate, oscillation and fermentation 2 days, are transferred to after fermentation ends at 32 DEG C
In centrifuge, 15min is centrifuged with 6000r/min, supernatant is taken, obtains freshwater mussel zymotic fluid, add and go into freshwater mussel zymotic fluid
Ionized water regulation substrate protein mass fraction is 5%, and regulation system pH is 9.5, then adds basic protein than 200U/g by enzyme bottom
Enzyme, 3h is digested at 50 DEG C, then be transferred in autoclave, the autoclaving 15min at 121 DEG C, load centrifugation after being cooled to room temperature
In machine, 15min is centrifuged with 6000r/min, takes supernatant liquor to be placed in freeze drying box, is freeze-dried at -20 DEG C
15h, freshwater mussel Gly-His-Lys are obtained, take 30g reduced sugars, 10g freshwater mussel Gly-His-Lys, add in 400mL deionized waters, stirred with 300r/min
20min, regulation system pH are 11, and are heated to 90 DEG C, are fitted into after reacting 3h in centrifuge, are centrifuged with 6000r/min
15min, take upper liquid to be placed in Rotary Evaporators, be evaporated to dryness at 50 DEG C, obtain freshwater mussel antioxidation active peptides.
Example 2
Take fresh freshwater mussel to shell, remove internal organ after wash 2 times, and be transferred to meat grinder rub after cross colloid mill, add deionized water,
Regulation to solid content is 8%, obtains mussel meat homogenate, takes 1.5kg mussel meats to be homogenized, 20g urea, 4g potassium hydrogen phosphates, 8g sodium chloride,
25min is stirred with 350r/min, and is transferred in autoclave, autoclaving 18min, obtains fermentation substrate at 123 DEG C, by inoculum concentration
3.5%（v/v）Bacillus subtilis is seeded in fermentation substrate, oscillation and fermentation 2 days at 35 DEG C, be transferred to after fermentation ends from
In scheming, with 7000r/min centrifuge 18min, take supernatant, obtain freshwater mussel zymotic fluid, into freshwater mussel zymotic fluid add go from
Sub- water regulation substrate protein mass fraction is 5%, and regulation system pH is 9.5, then adds basic protein than 200U/g by enzyme bottom
Enzyme, 3h is digested at 55 DEG C, then be transferred in autoclave, the autoclaving 18min at 123 DEG C, load centrifugation after being cooled to room temperature
In machine, 18min is centrifuged with 7000r/min, takes supernatant liquor to be placed in freeze drying box, is freeze-dried at -18 DEG C
18h, freshwater mussel Gly-His-Lys are obtained, take 50g reduced sugars, 15g freshwater mussel Gly-His-Lys, add in 600mL deionized waters, stirred with 350r/min
25min, regulation system pH are 11, and are heated to 95 DEG C, are fitted into after reacting 3.5h in centrifuge, are centrifuged with 7000r/min
18min, take upper liquid to be placed in Rotary Evaporators, be evaporated to dryness at 55 DEG C, obtain freshwater mussel antioxidation active peptides.
Example 3
Take fresh freshwater mussel to shell, remove internal organ after wash 3 times, and be transferred to meat grinder rub after cross colloid mill, add deionized water,
Regulation to solid content is 10%, obtains mussel meat homogenate, takes 2kg mussel meats to be homogenized, 30g urea, 6g potassium hydrogen phosphates, 10g sodium chloride,
30min is stirred with 400r/min, and is transferred in autoclave, autoclaving 20min, obtains fermentation substrate at 125 DEG C, by inoculum concentration
5.0%（v/v）Bacillus subtilis is seeded in fermentation substrate, oscillation and fermentation 3 days at 38 DEG C, be transferred to after fermentation ends from
In scheming, with 8000r/min centrifuge 20min, take supernatant, obtain freshwater mussel zymotic fluid, into freshwater mussel zymotic fluid add go from
Sub- water regulation substrate protein mass fraction is 6%, and regulation system pH is 10.0, then adds basic protein than 200U/g by enzyme bottom
Enzyme, 4h is digested at 60 DEG C, then be transferred in autoclave, the autoclaving 20min at 125 DEG C, load centrifugation after being cooled to room temperature
In machine, 20min is centrifuged with 8000r/min, takes supernatant liquor to be placed in freeze drying box, is freeze-dried at -15 DEG C
20h, freshwater mussel Gly-His-Lys are obtained, take 60g reduced sugars, 20g freshwater mussel Gly-His-Lys, add in 800mL deionized waters, stirred with 400r/min
30min, regulation system pH are 12, and are heated to 100 DEG C, are fitted into after reacting 4h in centrifuge, are centrifuged with 8000r/min
20min, take upper liquid to be placed in Rotary Evaporators, be evaporated to dryness at 60 DEG C, obtain freshwater mussel antioxidation active peptides.
Reference examples：The grape pip soft capsules of certain brand production.
The anti-oxidation peptide of example and reference examples is detected, specific detection is as follows：
The measure of reducing power：Lyophilized sample is accurately weighed, is configured to 1mg/mL sample liquid, takes 1.2mL sample liquids to delay with 3mL phosphate
Fliud flushing (0.2mol/L, pH6.6) mixes, and adds 3mL1% potassium ferricyanide solutions, and reaction solution is incubated 20min in 50 DEG C of water-baths, adds
3mL10%TCA solution, mix, 10min is centrifuged under the conditions of 3000r/min, take 2mL supernatants and 2mL distilled water to mix, add
0.5mL0.1%FeCl3 solution, reaction 10min is stored at room temperature, absorbance is measured at 700nm.
The measure of DPPH free radical scavenging activities：DPPH free radical scavenging activities determine with reference to YouLJ etc. method, and slightly repair
Change.3.94mgDPPH accurately is weighed, 100mL is settled to 95% ethanol, obtains 0.1mmol/LDPPH solution.
Specific testing result such as table 1.
Table 1
| Detection project | Example 1 | Example 2 | Example 3 | Reference examples |
| Reducing power | 2.5 | 2.0 | 1.5 | 1.0 |
| DPPH free radical scavenging activities（%） | 96.79 | 93.01 | 81.32 | 78.81 |
As shown in Table 1, the anti-oxidation peptide that prepared by the present invention has stronger radical scavenging activity and antioxidation activity.
Claims (7)
1. a kind of preparation method of freshwater mussel antioxidation active peptides, it is characterised in that specifically preparation process is：
（1）Take fresh freshwater mussel to shell, remove internal organ after wash, and be transferred to meat grinder rub after cross colloid mill, add deionized water
Solid content is adjusted, obtains mussel meat homogenate；
（2）Mussel meat homogenate, urea, potassium hydrogen phosphate, sodium chloride are taken, simultaneously autoclave sterilization is well mixed, obtains fermentation substrate；
（3）By inoculum concentration 2.5~5.0%（v/v）Bacillus subtilis is seeded in fermentation substrate and fermented, after fermentation ends from
Heart separation takes supernatant, obtains freshwater mussel zymotic fluid；
（4）Deionized water regulation substrate protein mass fraction and regulation system pH are added into freshwater mussel zymotic fluid, adds alkalescence
Protease hydrolyzed, centrifuged after sterilizing and take supernatant liquor, be freeze-dried to obtain freshwater mussel Gly-His-Lys；
（5）Reduced sugar and freshwater mussel Gly-His-Lys mixing addition are gone from water, regulation system pH, and heating response, after completion of the reaction from
The heart separates, and takes upper liquid to be evaporated to dryness, obtains freshwater mussel antioxidation active peptides.
A kind of 2. preparation method of freshwater mussel antioxidation active peptides as claimed in claim 1, it is characterised in that step（1）It is described
Solid content is 5~10%.
A kind of 3. preparation method of freshwater mussel antioxidation active peptides as claimed in claim 1, it is characterised in that step（2）It is described
Mussel meat homogenate, urea, potassium hydrogen phosphate, the parts by weight of sodium chloride are homogenized for 1000~2000 parts of mussel meats, 15~30 parts of urea,
3~6 parts of potassium hydrogen phosphates, 5~10 parts of sodium chloride.
A kind of 4. preparation method of freshwater mussel antioxidation active peptides as claimed in claim 1, it is characterised in that step（3）It is described
Fermentation process is oscillation and fermentation 2~3 days at 32~38 DEG C.
A kind of 5. preparation method of freshwater mussel antioxidation active peptides as claimed in claim 1, it is characterised in that step（4）It is described
Substrate protein mass fraction is 5~6%, and regulation system pH is 9.5~10.0, and alkali protease is added than 200U/g by enzyme bottom,
3~4h is digested at 50~60 DEG C.
A kind of 6. preparation method of freshwater mussel antioxidation active peptides as claimed in claim 1, it is characterised in that step（5）It is described
Reduced sugar is the one or more in glucose, lactose, maltose, galactolipin, xylose.
A kind of 7. preparation method of freshwater mussel antioxidation active peptides as claimed in claim 1, it is characterised in that step（5）It is described
Regulation system pH is 11~12, is heated to 90~100 DEG C of 3~4h of reaction.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108517343A (en) * | 2018-04-26 | 2018-09-11 | 厦门元之道生物科技有限公司 | A kind of preparation method of the anti-oxidant protein peptides of Porphyra yezoensis |
| CN109385457A (en) * | 2018-11-07 | 2019-02-26 | 四川大学 | A kind of preparation method of the giant salamander Mei Lade peptide with antioxidant activity |
| CN112544874A (en) * | 2020-11-19 | 2021-03-26 | 青岛农业大学 | Nutritional noodles added with fish polypeptide powder and preparation method thereof |
-
2017
- 2017-09-06 CN CN201710793754.0A patent/CN107365823A/en not_active Withdrawn
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108517343A (en) * | 2018-04-26 | 2018-09-11 | 厦门元之道生物科技有限公司 | A kind of preparation method of the anti-oxidant protein peptides of Porphyra yezoensis |
| CN108517343B (en) * | 2018-04-26 | 2021-02-02 | 厦门元之道生物科技有限公司 | Preparation method of porphyra yezoensis antioxidant protein peptide |
| CN109385457A (en) * | 2018-11-07 | 2019-02-26 | 四川大学 | A kind of preparation method of the giant salamander Mei Lade peptide with antioxidant activity |
| CN109385457B (en) * | 2018-11-07 | 2022-02-11 | 四川大学 | Preparation method of giant salamander Maillard peptide with antioxidant activity |
| CN112544874A (en) * | 2020-11-19 | 2021-03-26 | 青岛农业大学 | Nutritional noodles added with fish polypeptide powder and preparation method thereof |
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