CN109349415A - The preparation method of camellia pollen protein isolate - Google Patents
The preparation method of camellia pollen protein isolate Download PDFInfo
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- CN109349415A CN109349415A CN201811523683.3A CN201811523683A CN109349415A CN 109349415 A CN109349415 A CN 109349415A CN 201811523683 A CN201811523683 A CN 201811523683A CN 109349415 A CN109349415 A CN 109349415A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/006—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from vegetable materials
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Abstract
The invention discloses a kind of preparation methods of camellia pollen protein isolate, comprising the following steps: (1) takes camellia pollen, ungrease treatment；(2) step (1) degreasing camellia pollen is taken, broken wall treatment obtains pollen solution；(3) in the pollen solution obtained to step (2), it is 1:20-80 that deionized water adjustment solid-to-liquid ratio, which is added, adjusts pH to 7.5-12, constant temperature stirring, centrifuging and taking supernatant；(4) the supernatant pH to 3.0-5.0 that regulating step (3) is collected, is centrifugated out albumen, protein liquid pH is adjusted to 7.0 after washing, be freeze-dried to obtain camellia amyloid proteins.Preparation method of the present invention uses a variety of technology for broken wall, improves the yield of protein, wherein by examination and comparison protein yield and protein degree, enzymatic hydrolysis-ULTRASONIC COMPLEX technology for broken wall effect is best；The present invention, which prepares gained camellia amyloid proteins hydrolysate obtained after simulated gastrointestinal tract digests, has antifatigue oxidation resistant activity.
Description
Technical field
The invention belongs to the separation and extraction technologies of camellia pollen, more particularly to a kind of preparation side of camellia pollen protein isolate
Method.
Background technique
Camellia pollen refers to the pollen ball that honeybee acquires out of Tea anthers, and the annual 9-11 month is its production period, camellia
Powder fecund in Zhejiang, Anhui, Jiangsu, etc. ground.Fresh camellia pollen has special tea perfume breath, tastes taste slightly
Sweet tea, pollen particles are in Chinese red mostly.Camellia pollen have improve the immunity of the human body, cardioprotection blood pressure lowering, anti-senility etc.
Effect.Wherein protein rich in, unsaturated fatty acid, flavones, polyphenol, polysaccharide isoreactivity substance.Wherein pollen protein
Matter content is especially abundant, and average content is 20% or so.
Pollen has special cell wall, and outer wall is more hard, the dense matters structure such as sporopollenin, cellulose and pollenin
At pollen wall, high temperature and great pressure can not only be resisted, for physics, can be changed with corrosion-resistant by existing
It learns and the degradation of enzyme is all more stable.This layer of pollen wall is extremely hard, so when preparing pollen protein isolate, it need to be using broken
Wall processing.Domestic and international more typical wall-breaking method mainly has physical wall breaking and biological wall breaking method at present.And about wall-breaking method pair
The influence of the molecular weight and digestibility of prepared protein has not been reported.And it has no about camellia pollen protein isolate
Prepare relevant report.
Summary of the invention
Goal of the invention: in view of the above-mentioned defects in the prior art, it can be obtained the present invention provides one kind and be easier to digestion suction
The preparation method of the camellia pollen protein isolate of the camellia pollen protein isolate of receipts.
A kind of technical solution: preparation method of camellia pollen protein isolate of the present invention, comprising the following steps:
(1) camellia pollen, ungrease treatment are taken；
(2) step (1) degreasing camellia pollen is taken, broken wall treatment obtains pollen solution；
(3) in the pollen solution obtained to step (2), be added deionized water adjustment solid-to-liquid ratio be 1:20-80, adjust pH to
7.5-12, constant temperature stirring, centrifuging and taking supernatant；
(4) the supernatant pH to 3.0-5.0 that regulating step (3) is collected, is centrifugated out albumen, by protein liquid pH after washing
7.0 are adjusted to, camellia amyloid proteins is freeze-dried to obtain.
In step (1), the camellia pollen is placed in Soxhlet extractor degreasing, and degreasing solvent is petroleum ether (30-60 boiling range), rainbow
After inhaling 2-3 times, the degreasing pollen that takes out.
In step (2), the broken wall treatment is using in supersonic wave wall breaking processing, difference Poluo wall processing and enzymatic hydrolysis broken wall treatment
One or more kinds of modes carry out.Compound broken wall includes the modes such as the temperature difference-ultrasound, enzymatic hydrolysis-ultrasound.
Further, the supersonic wave wall breaking processing refers to: extracting degreasing camellia pollen, with the dispersion of 1:6-16 (g/mL) solid-to-liquid ratio
In deionized water, then stirring and dissolving is handled it using ultrasonic cell disruptor；At ultrasonic cell disruptor
Manage bar part are as follows: ultrasound intensity 200-600w, ultrasonic time 5-40 minutes, ultrasonic ETAD expected time of arrival and departure 0-5 seconds, ultrasound was closed time 0-5 seconds.
Further, the difference Poluo wall processing refers to: extracting degreasing camellia pollen is put in the subzero 80 DEG C of freezings 0- of subzero 4-
48h, is then added 50-100 DEG C of hot water with solid-liquid ratio 1:6-16 (g/mL), and stirring and dissolving is then placed in 25-90 DEG C of water-bath
Water-bath 1-12h.
Further, the enzymatic hydrolysis broken wall treatment refers to: extracting degreasing camellia pollen is dispersed in 1:6-16 (g/mL) solid-to-liquid ratio
In deionized water, stirring and dissolving, adjustment pH is 2.0-6.0, and complex enzyme, 25-65 DEG C of enzymatic hydrolysis is added, and enzymatic hydrolysis is put into 60- after the completion
Inactivate enzyme within 2-20 minutes in 100 DEG C of water-baths.
Wherein, the complex enzyme group becomes cellulase, pectase, zytase and papain.
Preferably, the total additional amount of the complex enzyme be 100g pollen protein add 1.2g complex enzyme, i.e., 1.2%；It is various
The proportion of enzyme is cellulase: pectase: zytase: papain=4:2:1:3.
Further, the temperature difference-ULTRASONIC COMPLEX broken wall refers to: by difference Poluo wall treated pollen solution, using
Ultrasonic cell disruptor continues to handle to it；Ultrasonic cell disruptor treatment conditions are as follows: ultrasound intensity 200-600w,
Ultrasonic time 5-40 minutes, ultrasonic ETAD expected time of arrival and departure 0-5 seconds, ultrasound was closed time 0-5 seconds.
Further, the enzymatic hydrolysis-ULTRASONIC COMPLEX broken wall refers to: the pollen solution after digesting broken wall treatment, uses
Ultrasonic cell disruptor continues to handle to it；Ultrasonic cell disruptor treatment conditions are as follows: ultrasound intensity 200-600w,
Ultrasonic time 5-40 minutes, ultrasonic ETAD expected time of arrival and departure 0-5 seconds, ultrasound was closed time 0-5 seconds.
The application preferably enzymatic hydrolysis-ULTRASONIC COMPLEX broken wall treatment mode.
In step (3), pH to 7.5-12 is adjusted using the NaOH solution of 0.1-1mol/L.
In step (3), the constant temperature stirring refers to stirs 10-100 minutes in 25-75 DEG C of thermostat water bath.
In step (3), the centrifugation, which refers to, to be centrifuged 5-30 minutes with centrifuge with the revolving speed of 2500-5000rpm/min.
In step (4), pH is adjusted to 3.0-5.0 with 0.1-1mol/L hydrochloric acid.
In step (4), the centrifugation refers to 2500-5000rpm/min centrifugation 5-30 minutes.
In step (4), it is centrifugated out albumen, is washed with deionized 1-5 times.
In step (4), pH is adjusted to 7.0 with the NaOH of 0.1-1mol/L by gained protein liquid.
Solid-to-liquid ratio unit described herein is g/mL.
The broken wall treatment of preparation method of the present invention is mainly handled using ultrasonication, temperature difference processing and complex enzyme hydrolysis,
Then camellia pollen protein isolate is prepared using isoelectric point precipitation.Preparation gained camellia pollen protein isolate is yellow powder, albumen
Matter content>90%, moisture content<10%, carbohydrate<0.5%；Digestibility is high, and molecular weight does not change.
The utility model has the advantages that being compared to the prior art, preparation method of the present invention uses a variety of technology for broken wall, improves protein
Yield, wherein by examination and comparison protein yield and protein degree, enzymatic hydrolysis-ULTRASONIC COMPLEX technology for broken wall effect is most
It is good；The present invention prepare gained camellia amyloid proteins hydrolysate obtained after simulated gastrointestinal tract digests have it is antifatigue anti-oxidant
Activity.
Detailed description of the invention
Fig. 1 is influence of the different broken wall modes to camellia pollen cell wall structure；
Fig. 2 is influence of the different broken wall modes to camellia pollen protein yield；
Fig. 3 is influence of the different broken wall modes to camellia pollen protein molecular weight；
Fig. 4 is influence of the different broken wall modes to camellia pollen cell wall structure.
Wherein, Control indicates control group (non-broken pollen), and U indicates that ultrasonication, T indicate that difference Poluo wall, E indicate
Broken wall is digested, T+U indicates that the temperature difference-ULTRASONIC COMPLEX broken wall, E+U indicate enzymatic hydrolysis-ULTRASONIC COMPLEX broken wall.
Specific embodiment
The application is explained in detail combined with specific embodiments below.
Raw material sources: camellia pollen --- purchased from Xinjiang Xin Yuan bee-keeping cooperative society.
Cellulase, pectase, zytase, papain are purchased from Shanghai Yuan Ye Biotechnology Co., Ltd.
Human umbilical vein endothelial cells (HUVEC) are purchased from Nanjing Bo Quan Science and Technology Ltd..
Embodiment 1
A kind of preparation method of camellia pollen protein isolate, comprising the following steps:
(1) pollen ungrease treatment；Camellia pollen is taken, is put into Soxhlet extractor, petroleum ether is added, is flowed back 2 times；
(2) pollen supersonic wave wall breaking is handled: the degreasing camellia pollen for taking step (1) to obtain, with 1:12 (g/mL) solid-liquid score
It dissipating in deionized water, stirring is completely dissolved pollen solution, then it handled using ultrasonic cell disruptor, if
Ultrasound intensity 400w is set, ultrasonic time 20 minutes, ultrasonic ETAD expected time of arrival and departure 2s, ultrasound closed time 2s；
(3) preparation of pollen protein isolate: the material water that deionized water adds to 1:40 is added in the pollen solution after broken wall treatment
Than adjusting pH to 10 with NaOH solution, being stirred 80 minutes in 55 DEG C of thermostat water bath, with centrifuge with 3000rpm/min
Revolving speed be centrifuged and 10 minutes and take out supernatant；PH is adjusted to 4.0 with hydrochloric acid by gained supernatant, then with centrifuge 3000rpm/
Min is centrifuged 10 minutes, isolates albumen, and be washed with deionized twice, and pH is adjusted to 7.0 with NaOH by gained protein liquid,
Camellia amyloid proteins is obtained after freeze-drying.
Embodiment 2
A kind of preparation method of camellia pollen protein isolate, step (2) are as follows: the processing of pollen difference Poluo wall: extracting degreasing camellia pollen,
It is put into -20 DEG C of refrigerators and freezes for 24 hours, 80 DEG C of hot water are then added with solid-liquid ratio 1:12 (g/mL), stirring is dissolved pollen, is put into
Water-bath 7h in 45 DEG C of water-baths.
Remaining operation is the same as embodiment 1.
Embodiment 3
A kind of preparation method of camellia pollen protein isolate, step (2) are as follows: pollen digests broken wall treatment: extracting degreasing camellia pollen,
In deionized water with the dispersion of 1:12 (g/mL) solid-to-liquid ratio, stirring dissolves pollen, and complex enzyme 1.2%, enzymatic hydrolysis is added in pH4.0
Temperature 45 C, enzyme ratio are cellulase: pectase: zytase: papain=4:2:1:3 is put into water-bath
Middle enzymatic hydrolysis 6h.Being put into 10 minutes in 80 DEG C of water-baths after the completion of enzymatic hydrolysis inactivates enzyme.
Remaining operation is the same as embodiment 1.
Embodiment 4
A kind of preparation method of camellia pollen protein isolate, step (2) are as follows: pollen difference Poluo wall carries out ultrasonic wave again later
Processing.
Remaining operation is the same as embodiment 1.
Embodiment 5
A kind of preparation method of camellia pollen protein isolate, step (2) are as follows: pollen digests broken wall, carries out ultrasonic wave again later
Processing.
Remaining operation is the same as embodiment 1.
Comparative example 1
A kind of preparation method of camellia pollen protein isolate, omits the broken wall treatment of step (2), and direct extracting degreasing camellia pollen is pressed
The step of according to embodiment 1 (3) processing.
Camellia amyloid proteins progressive energy interpretation of result to embodiment 1-5 and comparative example 1
1, influence of the different technology for broken wall to camellia pollen cell wall structure
The pollen solution after Example 1-5 step (2) broken wall treatment and comparative example 1 do not carry out broken wall treatment respectively
Camellia pollen observes influence of the different technology for broken wall to camellia pollen cell wall structure.As a result as shown in Figure 1, compared with the control group, breaking
Wall processing can destroy the structure of pollen cell wall.It compares between each technology for broken wall, enzymatic hydrolysis-ULTRASONIC COMPLEX technology for broken wall pair
The structure of pollen wall breaks ring maximum, the structure of cell wall can be destroyed completely, to be conducive to the release of protein.
2, influence of the different technology for broken wall to camellia amyloid proteins yield
Using the total protein content in Kjeldahl nitrogen determination pollen and freeze-dried powder, the calculating of yield according to the following equation:
Protein yield=(quality of total protein in quality/pollen of total protein in freeze-dried powder) × 100
As a result as shown in Fig. 2, sample of all samples through broken wall treatment relative to non-broken wall, albumen yield have centainly
The raising of degree.It compares between each technology for broken wall, when using enzymatic hydrolysis-ULTRASONIC COMPLEX technology for broken wall, protein yield reaches most
Greatly.
3, influence of the different technology for broken wall to camellia pollen molecular weight of albumen
Camellia pollen protein protomer and its molecular weight are analyzed using polyacrylamide gel electrophoresis (SDS-PAGE).Sample concentration
For 2mg/mL, applied sample amount is 20 μ L, and resolving gel concentration 12%, concentration gum concentration is 5%.Using current constant mode, glue electricity is concentrated
Flow control increases to 30mA in 15mA, separation gel electric current.After to be separated, glue is put into fixer and fixes 1h, then with examining
Mas bright blue dyeing liquor dyes 1h, and it is clear to band finally to be decolourized with destainer, is then taken pictures with gel imager.
As a result as shown in figure 3, supersonic wave wall breaking, difference Poluo wall, the temperature difference-ULTRASONIC COMPLEX broken wall are to the subunit of camellia amyloid proteins
Composition and molecular weight do not influence.And enzymatic hydrolysis broken wall and enzymatic hydrolysis-resulting protein of ULTRASONIC COMPLEX broken wall is used small molecule occur
The subunit of amount, this may be because used protease has occurred enzymatic hydrolysis to camellia pollen protein and makees during enzymatic hydrolysis broken wall
With.
4, the measurement of pollen protein isolate Vitro Digestibility
In vitro digestion: the pollen protein solution that configuration protein concentration is 4%, sample is with the revolving speed of 3000rpm/min
Centrifugation, takes supernatant spare.100mL supernatant is taken, PH is adjusted to 3, its water-solubility protein is added under 37 DEG C of water bath
The pepsin of amount 1%, and Gastric juice digestion sample is taken out after 37 DEG C of shaking table 2h, first time shaking table, pH is adjusted to 7, is added
Enter the trypsase of its water-solubility protein amount 1%, and the shaking table 2h at 37 DEG C, the sample digested is then put into 90 DEG C of water-baths
10min inactivation in pot.
The measurement of digestibility: 1mL trinitrobenzene sulfonic acid (TNBS) solution is added in the sample solution for taking 1mL to digest, then slowly
1mL distilled water is added as buffer, is put into 50 DEG C of water-baths and is protected from light water-bath 30min, 2mL terminate liquid (0.1mol/ is then added
L Na2SO3), it is eventually adding 5mL distilled water, using the cysteine of 2mg/mL as standard items, absorbance is measured at 340nm.
Protein degree=(content/pollen protein content of free amine group in enzymolysis liquid) × 100
The size of degree of hydrolysis is used to characterize the Vitro Digestibility of protein.
The degree of hydrolysis of camellia amyloid proteins is measured using in-vitro simulated pipe intestinal digesting system.As a result as shown in figure 4,
Compared with the control group, higher degree of hydrolysis is shown using technology for broken wall albumen obtained.It is wherein multiple using enzymatic hydrolysis-ultrasound
It closes technology for broken wall albumen obtained and shows highest degree of hydrolysis.
Embodiment 6
For camellia amyloid proteins preparation process referring to embodiment 5, the hydrolysis of camellia amyloid proteins, will referring in vitro digestion part
The sample solution of enzymatic hydrolysis obtains camellia pollen protolysate after being freeze-dried.
Mouse anti-reflecting fatigue experiment: mouse carries out treadmill acclimatization training in 1 week.Run duration is daily 5min, movement speed
Degree is incremented by 4m/min since 4m/min daily, and when increasing to 16m/min, movement velocity is not further added by, and finally maintains 16m/min,
Current strength 1.3mA.After mouse adapts to treadmill progress intensity acclimatization training in one week, selection can adapt to increasing intensity running
The mouse of machine training, is randomly divided into 6 groups by weight, every group 8, respectively control, camellia pollen, camellia amyloid proteins, camellia pollen egg
White hydrolysate 0.09,0.26,0.78g/kg group.Control group stomach-filling distilled water, sample sets continuous gavage difference sample, every mouse
Daily stomach-filling 0.1ml/10g.Free diet and water during test are weighed in weekly primary, and continuous gavage 6 days.Setting movement
Speed 16m/min, run duration 5min, record mouse are electrically shocked number, and 6 days weekly, 1 time a day.Statistical result such as 1 institute of table
Show.
Influence of the 1 camellia pollen protolysate of table to mouse running number of shocks and weight
#p < 0.05, compared with the control group；* p < 0.05, compared with camellia pollen group.
Conclusion: camellia pollen and camellia pollen protolysate are substantially reduced mouse and fall treadmill number, improve running training and draw
The fatigue risen, but weight is had no significant effect.
Embodiment 7
For camellia amyloid proteins preparation process referring to embodiment 5, the hydrolysis of camellia amyloid proteins, will referring in vitro digestion part
The sample solution of enzymatic hydrolysis obtains camellia pollen protolysate after being freeze-dried.
Anti-oxidant experiment: Human umbilical vein endothelial cells (HUVEC) are inoculated in 96 according to the density of 10,000 cells/wells
In orifice plate, it is placed in incubator and cultivates for 24 hours.Be divided into different disposal group: 1) control group: absorbing former culture medium, and 100 μ L are added in every hole
The culture medium of preheating；2) model of oxidative group: absorbing former culture medium, and the culture medium that hydrogen peroxide concentration is 0.3mM is added in every hole,
It is placed in incubator and acts on 1h.3) camellia pollen protolysate group: absorbing former culture medium, and every hole is added hydrogen peroxide concentration and is
The camellia pollen protolysate culture medium of 0.3mM and various concentration, final concentration of the 6.4 × 10 of camellia pollen protolysate-5,
3.2×10-4, 1.6 × 10-3, 0.8 × 10-3, 4.0 × 10-2G/mL is placed in incubator and acts on 1h.Absorb former culture medium, every hole
The culture medium and 10 μ L thiazolyl blues (MTT, 5mg/mL) of 90 μ L preheating is added, is placed in incubator and acts on 3h.MTT solution is absorbed,
150 μ L dimethyl sulphoxide solutions (DMSO) are added in every hole, are placed on shaking table and are protected from light effect 30min.570nm wave is measured with microplate reader
Absorbance under long, absorbance value is proportional with cell viability, and absorbance value is bigger, and cell viability is higher.Statistical result is shown in Table
2。
The influence for the HUVEC cell viability that 2 camellia pollen protolysate of table induces hydrogen peroxide
##p < 0.05, compared with the control group；*, p < 0.05p < 0.01 * *, compared with hydrogen peroxide group.
Conclusion: certain density hydrogen peroxide will lead to HUVEC peroxide injury, and result of study shows camellia amyloid proteins water
Solving object concentration is 3.2 × 10-4, 1.6 × 10-3, 0.8 × 10-3, 4.0 × 10-2G/mL is substantially reduced the peroxidating of hydrogen peroxide induction
Damage.
Claims (9)
1. a kind of preparation method of camellia pollen protein isolate, which comprises the following steps:
(1) camellia pollen, ungrease treatment are taken；
(2) step (1) degreasing camellia pollen is taken, broken wall treatment obtains pollen solution；
(3) in the pollen solution obtained to step (2), it is 1:20-80 that deionized water adjustment solid-to-liquid ratio, which is added, adjusts pH to 7.5-
12, constant temperature stirring, centrifuging and taking supernatant；
(4) the supernatant pH to 3.0-5.0 that regulating step (3) is collected, is centrifugated out albumen, is adjusted to protein liquid pH after washing
7.0, it is freeze-dried to obtain camellia amyloid proteins.
2. the preparation method of camellia pollen protein isolate according to claim 1, which is characterized in that described broken in step (2)
Wall processing is carried out using one or more of supersonic wave wall breaking processing, difference Poluo wall processing and enzymatic hydrolysis broken wall treatment mode.
3. the preparation method of camellia pollen protein isolate according to claim 2, which is characterized in that the broken wall treatment uses
Enzymatic hydrolysis-ULTRASONIC COMPLEX broken wall treatment.
4. the preparation method of camellia pollen protein isolate according to claim 2, which is characterized in that at the supersonic wave wall breaking
Reason refers to: taking camellia pollen, in deionized water with the dispersion of 1:6-16 solid-to-liquid ratio, then stirring and dissolving uses supersonic cell powder
Broken machine handles it；Ultrasonic cell disruptor treatment conditions are as follows: ultrasound intensity 200-600w, ultrasonic time 5-40 points
Clock, ultrasonic ETAD expected time of arrival and departure 0-5 seconds, ultrasound was closed time 0-5 seconds.
5. the preparation method of camellia pollen protein isolate according to claim 2, which is characterized in that the difference Poluo wall processing
Refer to: taking camellia pollen, be put in the subzero 80 DEG C of freezings 0-48h of subzero 4-, 50-100 DEG C of hot water is then added with solid-liquid ratio 1:6-16,
Stirring and dissolving is then placed in water-bath 1-12h in 25-90 DEG C of water-bath.
6. the preparation method of camellia pollen protein isolate according to claim 2, which is characterized in that the enzymatic hydrolysis broken wall treatment
Refer to: taking camellia pollen, in deionized water with the dispersion of 1:6-16 solid-to-liquid ratio, stirring and dissolving, adjustment pH is 2.0-6.0, is added multiple
Synthase, 25-65 DEG C of enzymatic hydrolysis, enzymatic hydrolysis is put into 2-20 minutes in 60-100 DEG C of water-bath after the completion inactivates enzyme.
7. the preparation method of camellia pollen protein isolate according to claim 6, which is characterized in that the complex enzyme group becomes
Cellulase, pectase, zytase and papain.
8. the preparation method of camellia pollen protein isolate according to claim 1, which is characterized in that in step (3), the perseverance
Temperature stirring refers to stirs 10-100 minutes in 25-75 DEG C of thermostat water bath；The centrifugation refers to centrifuge with 2500-
The revolving speed of 5000rpm/min is centrifuged 5-30 minutes.
9. the preparation method of camellia pollen protein isolate according to claim 1, which is characterized in that in step (4), it is described from
The heart refers to that 2500-5000rpm/min is centrifuged 5-30 minutes.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115251224A (en) * | 2022-04-29 | 2022-11-01 | 南京中医药大学 | Preparation method of bee pollen protein |
| CN115386611A (en) * | 2022-10-31 | 2022-11-25 | 山东睿鹰制药集团有限公司 | Peony pollen protein polypeptide and preparation method and application thereof |
-
2018
- 2018-12-13 CN CN201811523683.3A patent/CN109349415A/en active Pending
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| 杨天炯: "《蒙山茶飞跃历程》", 31 August 2018 * |
| 杨文超等: "莲花蜂花粉酶解破壁工艺条件优化", 《食品科学》 * |
| 范三红等: "油松花粉蛋白提取工艺的优化", 《食品工业科技》 * |
| 董捷等: "有机溶剂联合酶法处理对油菜花粉萌发孔通透性改善的研究", 《食品科学》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115251224A (en) * | 2022-04-29 | 2022-11-01 | 南京中医药大学 | Preparation method of bee pollen protein |
| CN115386611A (en) * | 2022-10-31 | 2022-11-25 | 山东睿鹰制药集团有限公司 | Peony pollen protein polypeptide and preparation method and application thereof |
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Application publication date: 20190219 |