CN104017850B - Method and the application of the little peptide of Paeonia suffruticosa of the little peptide of Paeonia suffruticosa prepared by a kind of seed of Flos Moutan grouts - Google Patents
Method and the application of the little peptide of Paeonia suffruticosa of the little peptide of Paeonia suffruticosa prepared by a kind of seed of Flos Moutan grouts Download PDFInfo
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- CN104017850B CN104017850B CN201410294944.4A CN201410294944A CN104017850B CN 104017850 B CN104017850 B CN 104017850B CN 201410294944 A CN201410294944 A CN 201410294944A CN 104017850 B CN104017850 B CN 104017850B
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Abstract
The invention discloses a kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, the method be extract oil with oil seed of Flos Moutan after the seed of Flos Moutan grouts that obtain as raw material, add water, trypsin, papain, subtilisin and alkaline 2709 protease and/or neutral Neutrase protease carry out mixed hydrolysis and makes the little peptide of Paeonia suffruticosa function.The weight ratio of described trypsin, papain, subtilisin, alkaline 2709 protease and neutral Neutrase protease is 1:0.8 1.2:0.8 1.2:0 1.2:0 1.2.The method that the present invention provides is quick, easy, mild condition, the little peptide of Paeonia suffruticosa prepared remains the function of seed of Flos Moutan to greatest extent, nutrition natural, high, easily absorbs, overcome the problem that vegetable protein is difficult to digest and assimilate, and the multiple biological activity such as antioxidation, antibacterial and antitumor is notable.
Description
Technical field
The invention belongs to agricultural byproducts processing and reutilization technology field, be specifically related to one seed of Flos Moutan grouts and prepare male
The method of red little peptide and the application of the little peptide of Paeonia suffruticosa.
Background technology
Peony seed oil contains multiple unsaturated fatty acid, the rarest odd-numbered fatty acid, is that one has important opening
The edible oil and fat source of value of making an offer.On March 22nd, 2011, Ministry of Health of the People's Republic of China approval from phoenix Paeonia suffruticosa (Paeonia ostii T.Hong et J.X.Zhang) and Paeonia papaveracea (Paeonia rockii) seed core squeeze through transmission from one meridian to another, decolour, de-
The peony seed oil that the technique such as smelly is made can produce as new resource food and edible [Ministry of Health of the People's Republic of China announces,
2011 No. 9: Ministry of Public Health about approval Acer Truncatum Buge seed oil and peony seed oil as the bulletin of new resource food].This will be for male
The biggest facilitation is played in production and the use of red seed oil.Due to the high nutritive value of peony seed oil, Paeonia suffruticosa used by China's oil
Plantation amount gradually increasing, the yield of oil seed of Flos Moutan increases the most year by year.According to current yield, oil seed of Flos Moutan yield is
30000 tons, be 30% calculating according to oil yield, will produce the seed of Flos Moutan grouts of about 10,000 tons after oil expression, these seed of Flos Moutan grouts
Exploitation would is that an extremely important and research topic for arduous task.
Summary of the invention
It is an object of the invention to provide a kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, the method is quick, easy,
Mild condition, the little peptide of Paeonia suffruticosa prepared remains the function of seed of Flos Moutan to greatest extent, nutrition natural, high, easily absorbs, gram
Take the problem that vegetable protein is difficult to digest and assimilate, and the multiple biological activity such as antioxidation, antibacterial and antitumor is notable.
The present invention realizes above-mentioned purpose and the technical scheme is that the side of the little peptide of Paeonia suffruticosa prepared by a kind of seed of Flos Moutan grouts
Method, the seed of Flos Moutan grouts obtained after extracting oil with oil seed of Flos Moutan, as raw material, add water, trypsin, papain, hay bar
Mycoproteinase and alkaline 2709 protease and/or neutral Neutrase protease carry out mixed hydrolysis, and to make Paeonia suffruticosa function little
Peptide.
Described trypsin, papain, subtilisin, alkaline 2709 protease and neutral Neutrase
The weight ratio of protease is 1:0.8-1.2:0.8-1.2:0-1.2:0-1.2.
The described method preparing the little peptide of Paeonia suffruticosa with seed of Flos Moutan grouts, specifically comprises the following steps that
1) take oil seed of Flos Moutan raw material, after shelling, obtain seed of Flos Moutan core, standby；The seed of Flos Moutan core aqueous alkali obtained is soaked
Bubble remove impurity, then wash 3-4 time with clear water, dry parch after moisture, then use milling process squeezing to remove oils and fats, obtain seed of Flos Moutan
Grouts, standby；
2) 60-100 mesh sieve pulverized by seed of Flos Moutan grouts step 1) obtained, and is then added in reactor, adds it
The water of weight 4-8 times stirs 15-20min at temperature is 50-70 DEG C, and regulation pH value is 6.4-7.5, obtains material liquid, standby；
3) by step 2) material liquid that obtains is warming up to 110-120 DEG C, and insulation 15-30 min carries out sterilizing, and sterilizing terminates
After be cooled to 40-50 DEG C, add and cultivate ripe mixing protease, and to make its mass percent in material liquid be 1.0-
8.0%, mixing protease by weight ratio be the trypsin of 1:0.8-1.2:0.8-1.2:0-1.2:0-1.2, papain,
Subtilisin, alkaline 2709 protease and neutral Neutrase protease mixing composition, control temperature and be 40-50 DEG C,
Mixing speed is 100-200 r/min, hydrolyzes 48-96h under the conditions of filtrated air, and hydrolysis is warming up to 110-120 DEG C after terminating
Carrying out high-temperature inactivation, filter cleaner, the filtrate obtained is the nutritional solution containing the little peptide of Paeonia suffruticosa.
Filtrate reduced in volume step 3) obtained to volume is the 1/4-1/3 of original volume, spray-dried prepared Paeonia suffruticosa
Little Gly-His-Lys.
The pH value of the aqueous alkali of described step 1) remove impurity is 8.5-9.5.
Described step 2) regulate pH value is 20% sodium hydroxide solution.
Described step 3) uses plate-and-frame filtration or membrane filtration to filter.
The described little peptide of Paeonia suffruticosa prepared with seed of Flos Moutan grouts is in the application of antioxidation, antitumor and antibiosis.
Beneficial effects of the present invention
The preparation method that one, the present invention provide is quick, easy, and mild condition, easy to operate, applicable scale is prepared male
The red little peptide of function, makes full use of compound protease to hydrolyze seed of Flos Moutan cake protein, and the little peptide molecular weight of Paeonia suffruticosa prepared is relatively
Little, it is evenly distributed, remains the function of seed of Flos Moutan to greatest extent, nutrition natural, high, easily absorb, overcome vegetable protein difficult
With the problem digested and assimilated, and the multiple biological activity such as antioxidation, antibacterial and antitumor is notable.
Garbage after two, the present invention makes full use of seed of Flos Moutan oil expression, by the research and development of seed of Flos Moutan grouts and recycling,
Utilization is researched and developed for seed of Flos Moutan grouts and will play certain facilitation, the research of deep processed product that Paeonia suffruticosa is correlated with
Exploitation has reference.
Detailed description of the invention
A kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, the seed of Flos Moutan grouts obtained after extracting oil with oil seed of Flos Moutan
For raw material, add water, trypsin, papain, subtilisin and alkaline 2709 protease and/or neutrality
Neutrase protease carries out mixed hydrolysis and makes the little peptide of Paeonia suffruticosa function.
Described trypsin, papain, subtilisin, alkaline 2709 protease and neutral Neutrase
The weight ratio of protease is 1:0.8-1.2:0.8-1.2:0-1.2:0-1.2.
The described method preparing the little peptide of Paeonia suffruticosa with seed of Flos Moutan grouts, specifically comprises the following steps that
1) take oil seed of Flos Moutan raw material, after shelling, obtain seed of Flos Moutan core, standby；The seed of Flos Moutan core aqueous alkali obtained is soaked
Bubble remove impurity, then wash 3-4 time with clear water, dry parch after moisture, then use milling process squeezing to remove oils and fats, obtain seed of Flos Moutan
Grouts, standby；
2) 60-100 mesh sieve pulverized by seed of Flos Moutan grouts step 1) obtained, and is then added in reactor, adds it
The water of weight 4-8 times stirs 15-20min at temperature is 50-70 DEG C, and regulation pH value is 6.4-7.5, obtains material liquid, standby；
3) by step 2) material liquid that obtains is warming up to 110-120 DEG C, and insulation 15-30 min carries out sterilizing, and sterilizing terminates
After be cooled to 40-50 DEG C, add and cultivate ripe mixing protease, and to make its mass percent in material liquid be 1.0-
8.0%, mixing protease by weight ratio be the trypsin of 1:0.8-1.2:0.8-1.2:0-1.2:0-1.2, papain,
Subtilisin, alkaline 2709 protease and neutral Neutrase protease mixing composition, control temperature and be 40-50 DEG C,
Mixing speed is 100-200 r/min, hydrolyzes 48-96h under the conditions of filtrated air, and hydrolysis is warming up to 110-120 DEG C after terminating
Carrying out high-temperature inactivation, filter cleaner, the filtrate obtained is the nutritional solution containing the little peptide of Paeonia suffruticosa.
Filtrate reduced in volume step 3) obtained to volume is the 1/4-1/3 of original volume, spray-dried prepared Paeonia suffruticosa
Little Gly-His-Lys.
The pH value of the aqueous alkali of described step 1) remove impurity is 8.5-9.5.
Described step 2) regulate pH value is 20% sodium hydroxide solution.
Described step 3) uses plate-and-frame filtration or membrane filtration to filter.
The described little peptide of Paeonia suffruticosa prepared with seed of Flos Moutan grouts is in the application of antioxidation, antitumor and antibiosis.
Below in conjunction with specific embodiment, the present invention will be further described:
Embodiment 1:
A kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, specifically comprises the following steps that
1) take Paeonia ostii Paeonia suffruticosa seed raw material 20, after shelling, obtain seed of Flos Moutan core, standby；Seed of Flos Moutan core pH that will obtain
Value is 9.5 alkaline soak remove impurity, then washs 4 times with clear water, dries parch after moisture, then uses milling process squeezing to go oil removing
Fat, obtains seed of Flos Moutan grouts 8.7, standby；
2) take the seed of Flos Moutan grouts 1 that step 1) obtains, pulverized 100 mesh sieves, be then added in reactor, add it
The water that weight is 8 times stirs 20min at temperature is 70 DEG C, is 6.4-7.5 with 20% sodium hydroxide solution regulation pH value, obtains former
Feed liquid, standby；
3) by step 2) material liquid that obtains is warming up to 120 DEG C, is incubated 30 min and carries out sterilizing, and sterilizing is cooled to after terminating
50 DEG C, add and cultivate ripe mixing protease, and to make its mass percent in material liquid be 8.0%, mixing protease by
Weight ratio is the trypsin of 1:1.2:1.2:1.2, papain, subtilisin, alkaline 2709 protease mixing
Composition, controlling temperature is 50 DEG C, and mixing speed is 200 r/min, hydrolyzes 96h under the conditions of filtrated air, and hydrolysis rises after terminating
Temperature carries out high-temperature inactivation to 110-120 DEG C, uses plate-and-frame filtration slagging-off, and the filtrate obtained is the nutrition containing the little peptide of Paeonia suffruticosa
Liquid.
Filtrate reduced in volume step 3) obtained to volume is the 1/3 of original volume, the spray-dried little peptide of prepared Paeonia suffruticosa
Powder 120g.
Embodiment 2:
A kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, specifically comprises the following steps that
1) take paeonia rockii seed raw material 10, after shelling, obtain seed of Flos Moutan core, standby；Seed of Flos Moutan core pH that will obtain
Value is 8.5 alkaline soak remove impurity, then washs 3 times with clear water, dries parch after moisture, then uses milling process squeezing to go oil removing
Fat, obtains seed of Flos Moutan grouts 3.6, standby；
2) take the seed of Flos Moutan grouts 1 that step 1) obtains, pulverized 60 mesh sieves, be then added in reactor, add it
The water that weight is 4 times stirs 15min at temperature is 50 DEG C, is 7.0 with 20% sodium hydroxide solution regulation pH value, obtains material liquid,
Standby；
3) by step 2) material liquid that obtains is warming up to 110 DEG C, is incubated 15 min and carries out sterilizing, and sterilizing is cooled to after terminating
40 DEG C, add and cultivate ripe mixing protease, and to make its mass percent in material liquid be 1.0%, mixing protease by
Weight ratio is the trypsin of 1:1.2:1.2:1.2, papain, subtilisin and neutral Neutrase albumen
Enzyme mixing composition, controlling temperature is 40 DEG C, and mixing speed is 100 r/min, hydrolyzes 48h, hydrolysis knot under the conditions of filtrated air
Being warming up to 110 DEG C after bundle and carry out high-temperature inactivation, use membrane filtration slagging-off, the filtrate obtained is the nutrition containing the little peptide of Paeonia suffruticosa
Liquid.
Filtrate reduced in volume step 3) obtained to volume is the 1/4 of original volume, the spray-dried little peptide of prepared Paeonia suffruticosa
Powder 108g.
Embodiment 3:
A kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, specifically comprises the following steps that
1) take the Paeonia ostii Paeonia suffruticosa with arbitrary proportion mixing and paeonia rockii seed raw material 15, after shelling, obtain seed of Flos Moutan
Core, standby；The alkaline soak remove impurity that seed of Flos Moutan core pH value is 9.0 that will obtain, then wash 4 times with clear water, after drying moisture
Parch, then uses milling process squeezing to remove oils and fats, obtains seed of Flos Moutan grouts 11.6, standby；
2) take the seed of Flos Moutan grouts 1 that step 1) obtains, pulverized 80 mesh sieves, be then added in reactor, add it
The water that weight is 6 times stirs 18min at temperature is 60 DEG C, is 6.4 with 20% sodium hydroxide solution regulation pH value, obtains material liquid,
Standby；
3) by step 2) material liquid that obtains is warming up to 115 DEG C, is incubated 22 min and carries out sterilizing, and sterilizing is cooled to after terminating
45 DEG C, add and cultivate ripe mixing protease, and to make its mass percent in material liquid be 4.5%, mixing protease by
Weight ratio is the trypsin of 1:1:1:1:1, papain, subtilisin, alkaline 2709 protease and neutrality
Neutrase protease mixing composition, controlling temperature is 45 DEG C, and mixing speed is 150 r/min, at filtrated air Water Under
Solving 80h, hydrolysis is warming up to 115 DEG C and carries out high-temperature inactivation after terminating, use plate-and-frame filtration slagging-off, and the filtrate obtained is containing male
The nutritional solution of red little peptide.
Filtrate reduced in volume step 3) obtained to volume is the 3/10 of original volume, the spray-dried little peptide of prepared Paeonia suffruticosa
Powder 107g.
Active testing is tested
1. antitumor activity
1) test material
Hyclone, 96 well culture plates, calorstat, RPM1640 culture fluid.
2) method
Peptide little to the Paeonia suffruticosa obtained in above-described embodiment carries out cytotoxic activity test, and the method for employing is mtt assay.
3) test procedure
(1) take the logarithm trophophase cell, with the RPM1640 culture fluid containing 10% hyclone, make single cell suspension 5 ×
104Individual/mL, is added to this suspension in 96 well culture plates, and every hole adds 1 × 104Individual cell.After cultivating 24 h, add tested little peptide
Concentration is respectively 30 μ g/mL, 10 μ g/mL.
(2) by flat board at 37 DEG C, containing 5%CO2The calorstat of air and 100% humidity is hatched 3 days, remove culture fluid.
(3) the RPM1640 culture fluid of MTT serum-free is made into 1 mg/mL solution, and every hole adds 100 μ L, 37 DEG C of incubations 4
H, makes MTT be reduced to first.
(4) the sucking-off supernatant, adds 150 μ L DMSO and makes first dissolve, and surveys OD value at ELISA instrument 570 nm.
4) result of the test
Table 1 Paeonia suffruticosa little polypeptide cell cytotoxic activity experimental data (n=3)
2. antioxidant activity research
1) test material
Paeonia suffruticosa function little peptide powder (preparation method is shown in embodiment 1).
Sodium nitrite, aluminum nitrate, sodium hydroxide, 1,1-diphenyl-2-picryl phenylhydrazine (DPPH), reduced form nicotinoyl amine gland are fast
Nicotinamide adenine dinucleotide (NADH), azophenlyene methosulfate (PMS), chlorination nitro tetrazole (NBT), dehydrated alcohol, hydrogen peroxide, 2,6-
Toluene di-tert-butyl phenol (BHT), salicylic acid, trishydroxymethylaminomethane (Tris), trichloroacetic acid (TCA), sulfur are for barbital
Acid (TBA), hydrochloric acid (HC1) are analytical pure, soybean lecithin (> 90%).
2) method
(1) ultra-oxygen anion free radical (O is removed- 2) assay method
Little for Paeonia suffruticosa peptide is diluted to different Concentraton gradient, respectively with the Tris-HCl buffer of pH8.0, concentration 50mol/L
Taking the 1.5mL little peptide solution of variable concentrations Paeonia suffruticosa, (300 μm ol/L, with the Tris-of pH8.0 to be then sequentially added into 0.5mL NBT
HCl buffer), 0.5mL NADH (468 μm ol/L, with the Tris-HCl buffer of pH8.0), 0.5mL PMS
(60 μm ol/L, with the Tris-HCl buffer of pH8.0), after mixing in 25 DEG C water-bath 5 minutes, take out at wavelength 560
Measuring absorbance at nm, blank group replaces stilbene class solution with buffer.
E(O- 2) (%)=(1-)×100 (1)
In formula: E (O- 2) it is that the little peptide of Paeonia suffruticosa is to O- 2Clearance rate (%)；A1For stilbene class absorbance；A0For Blank absorbance
Degree.
(2) assay method of DPPH free radical is removed
Little for Paeonia suffruticosa peptide is diluted to different Concentraton gradient, respectively takes the solution of the 2 above-mentioned concentration of mL in test tube, add 2
ML concentration is the DPPH solution of 0.04mg/mL, mix homogeneously, reacts 20 minutes, 3500 r/min centrifugations 10 minutes, takes
Its absorbance surveyed at wavelength 517 nm by clear liquid is Ai；The another little peptide solution of Paeonia suffruticosa of the above-mentioned concentration of 2mL that respectively takes is in test tube, respectively
Add dehydrated alcohol 2 mL, react 20 minutes, 3500 r/min centrifugations 10 minutes, take supernatant and survey at wavelength 517nm
Its absorbance is Aj；Reacting as reference using 2 mL 0.04 mg/mL DPPH and 2 mL dehydrated alcohol, its absorbance is designated as A0。
E (DPPH) (%)=(1-)×100 (2)
In formula: E (DPPH) is the Paeonia suffruticosa little peptide clearance rate (%) to DPPH free radical；A0For 2mL DPPH solution+2mL without
The absorbance of water-ethanol；AiAbsorbance for the 2mL DPPH solution little peptide solution of+2mL Paeonia suffruticosa；AjMale for 2mL dehydrated alcohol+2mL
The absorbance of red little peptide.
(3) assay method of scavenging hydroxyl (OH)
Little for Paeonia suffruticosa peptide distilled water is configured to variable concentrations gradient, respectively takes the solution of the above-mentioned concentration of 2mL, be sequentially added into
The FeSO of 2mL 6 mmol/L4, the H of 2 mL 6 mmol/L2O2, stand 10 minutes after mixing, add 2mL 6 mmol/L water
Poplar acid, mixing, stand 30 minutes, at wavelength 510 nm, survey its absorbance be designated as Ai, when with distilled water replacement salicylic acid
Absorbance is designated as Aj.Blank group replaces Paeonia suffruticosa little peptide solution absorbance to be designated as A with distilled water0。
E (OH) (%)=(1-)×100
In formula: E (OH) is the clearance rate (%) of hydroxy radical；AiAbsorbance for the little reactive polypeptide of Paeonia suffruticosa；A0Inhale for blank
Luminosity；AjThe absorbance of stilbene class during for participating in reaction without salicylic acid.
3) result
(1) ultra-oxygen anion free radical (O is removed2 -·)
The little peptide of table 2 Paeonia suffruticosa removes ultra-oxygen anion free radical (O2 -) activity
As shown in Table 2, in the range of selected mass concentration, little peptide compounds has the energy removing ultra-oxygen anion free radical
Power.Along with the increase of mass concentration, the scavenging action to ultra-oxygen anion free radical strengthens, and presents dose-effect relationship, general effect
More consistent than BHT.
(2) DPPH free radical is removed
The little peptide of table 3 Paeonia suffruticosa removes DPPH free radical activity
DPPH free radical is the most stable a kind of free radical centered by nitrogen, and its effect removed is shown test medicine
Have and reduce hydroxy radical, alkane free radical or the valid density of peroxy radical and interrupt the effect of lipid peroxidation chain reaction.
As shown in Table 3, little peptide compounds shows the ability of the strongest removing DPPH free radical.
(3) scavenging hydroxyl (OH)
Table 4 Paeonia suffruticosa little peptide scavenging hydroxyl (OH) activity
Hydroxyl radical free radical is the strongest oxidant, the most almost can react with all cells composition, right
Body harm is the biggest.As shown in Table 4, Paeonia suffruticosa little peptide removing OH effect is compared with BHT and is slightly stronger.
3. antibacterial activity test
1) test material
The little peptide of Paeonia suffruticosa (three groups of little peptides of Paeonia suffruticosa that above-mentioned three groups of embodiments obtain, respectively little peptide 1, little peptide 2, little peptide 3).
Bacterial strain: staphylococcus aureusS. aureas, streptococcus uberisS. uberis, bacillus subtilisBacillus subtilis, escherichia coliE. agalactiaeThered is provided by The First Affiliated Hospital of Henan University of Science and Technology.
2) method
The mensuration of minimum inhibitory concentration (MIC).Employing document [Li Jianqiang, Li Liujin. veterinary microbiology test is real
Practise instruct [M]. Xi'an: Shaanxi science tech publishing house, 1999:29-30] in test tube turbidimetry, by 10 band tampons
Sterilizing test tubes is numbered, and No. 1 pipe adds cultured solution of broth 1.5 mL, and remaining each pipe adds 1 mL.Take 10 mg/mL sample solutions (molten
Agent is the mixed liquor of V (methanol): V (water)=1: 1) 0.5 mL adds in No. 1 pipe and mixes, then take 1 mL from No. 1 pipe
Adding No. 2 pipes, such serial dilution to No. 8 is managed, and No. 8 pipes discard 1 mL meat soup Han medicinal liquid.Draw 50mL bacteria containing amount to be equivalent to
Bacterial suspension (bacterial concentration 1.5 hundred million/mL) to 1～No. 9 pipe of Maxwell opacity tube the 1st pipe 1/2,50 mL meat added by No. 10 pipes
Soup.Put 37 DEG C and cultivate taking-up observation turbidity after 12～15 h.No. 9 pipes are muddy positive control pipe, and No. 10 pipes are for muddy cloudy
Property control tube.Manage limpid and that each pipe is consistent with No. 9 pipe turbidity thereafter test tube than No. 9, its medicament contg is minimum antibacterial
Concentration.
3) result of the test
Research finds that the little peptide of Paeonia suffruticosa is to gram positive bacteria (staphylococcus aureus, streptococcus uberis, bacillus subtilis
Bacterium) and gram negative bacteria (escherichia coli) be respectively provided with good inhibiting effect, and drug effect is preferable.Experimental result is such as
Under:
Table 5 Paeonia suffruticosa little peptide minimum inhibitory concentration (MIC)
It is more than the preferred embodiment of the present invention, but scope is not limited to this, every without prejudice to this
Bright principle, spirit ... all at the row of protection of the present invention.
Claims (4)
1. the method preparing the little peptide of Paeonia suffruticosa with seed of Flos Moutan grouts, it is characterised in that: specifically comprise the following steps that
1) take oil seed of Flos Moutan raw material, after shelling, obtain seed of Flos Moutan core, standby；The seed of Flos Moutan core alkaline soak obtained is removed
Miscellaneous, the pH value of the alkali liquor of remove impurity is that 8.5-9.5 washs 3-4 time with clear water again, dries parch after moisture, then uses milling process
Oils and fats is removed in squeezing, obtains seed of Flos Moutan grouts, standby；
2) 60-100 mesh sieve pulverized by seed of Flos Moutan grouts step 1) obtained, and was then added in reactor, added its weight
The water of 4-8 times stirs 15-20min at temperature is 50-70 DEG C, and regulation pH value is 6.4-7.5, obtains material liquid, standby；
3) by step 2) material liquid that obtains is warming up to 110-120 DEG C, and insulation 15-30 min carries out sterilizing, and sterilizing is the coldest after terminating
But to 40-50 DEG C, add and cultivate ripe mixing protease, and to make its mass percent in material liquid be 1.0-8.0%,
Described mixing protease is the trypsin of 1:0.8-1.2:0.8-1.2:0-1.2:0-1.2, papain, withered by weight ratio
Grass Bacillus protease, alkaline 2709 protease and neutral Neutrase protease mixing composition, control temperature and be 40-50 DEG C, stir
Mixing speed is 100-200 r/min, hydrolyzes 48-96h under the conditions of filtrated air, and hydrolysis is warming up to 110-120 DEG C after terminating and enters
Row high-temperature inactivation, filter cleaner, the filtrate obtained is the nutritional solution containing the little peptide of Paeonia suffruticosa；
Filtrate reduced in volume step 3) obtained to volume is the 1/4-1/3 of original volume, the spray-dried little peptide of prepared Paeonia suffruticosa
Powder.
A kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, it is characterised in that: described step
Rapid 2) regulation pH value is 20% sodium hydroxide solution.
A kind of method that the little peptide of Paeonia suffruticosa prepared by seed of Flos Moutan grouts, it is characterised in that: described step
Rapid 3) plate-and-frame filtration or membrane filtration is used to filter.
4. the little peptide of Paeonia suffruticosa prepared with seed of Flos Moutan grouts as claimed in claim 1 application in terms of antioxidation.
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| CN108823273B (en) * | 2018-07-09 | 2020-12-18 | 齐鲁工业大学 | Peony seed meal polypeptide with antioxidant activity and preparation method and application thereof |
| CN110218755A (en) * | 2019-05-23 | 2019-09-10 | 世堃堂(广东)生物科技有限公司 | A kind of preparation method of hypoglycemic tree peony active peptide |
| CN110257460A (en) * | 2019-05-23 | 2019-09-20 | 世堃堂(广东)生物科技有限公司 | A kind of preparation method of the tree peony protein peptides of blood pressure lowering |
| CN111557450A (en) * | 2020-05-27 | 2020-08-21 | 许瀛瑄 | Cordyceps sinensis and peony oligopeptide lipid-lowering and anti-aging nutritional powder |
| CN112920376A (en) * | 2021-01-29 | 2021-06-08 | 深圳市中南活力实业股份有限公司 | Moisture-absorbing quick-drying water-based antibacterial polyurethane finishing agent and preparation method and application thereof |
| CN117363678B (en) * | 2023-10-10 | 2024-07-09 | 山东派恩生物科技有限公司 | Preparation method of peony fresh flower extracting solution |
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| JP4707401B2 (en) * | 2004-02-16 | 2011-06-22 | 株式会社山田養蜂場本社 | Antioxidant peptides from royal jelly |
| CN103493968B (en) * | 2013-09-23 | 2014-09-03 | 河南科技大学 | Preparation method of high-purity peony seed protein powder for oil |
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