CN109939051A - A kind of mulberries fermentation magma and its preparation method and application - Google Patents
A kind of mulberries fermentation magma and its preparation method and application Download PDFInfo
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- CN109939051A CN109939051A CN201910338889.7A CN201910338889A CN109939051A CN 109939051 A CN109939051 A CN 109939051A CN 201910338889 A CN201910338889 A CN 201910338889A CN 109939051 A CN109939051 A CN 109939051A
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Abstract
The invention belongs to field of biotechnology, and in particular to a kind of mulberries fermentation magma and its preparation method and application.Mulberries fermentation magma is by the way that mulberries, water and zymocyte liquid are mixed to get initial system, carry out fermented and cultured acquisition to initial system.In zymocyte liquid, strain is bacillus amyloliquefaciens LX-12 (Bacillus amyloliquefaciens LX-12), deposit number CCTCC NO:M 2015234.The present invention uses microbe fermentation method, and preparation method is easy, remains the whole components such as mulberries polysaccharide, resveratrol, anthocyanin and its activity；The content of the substances such as mulberries polyphenol, flavones in mulberries magma is also improved simultaneously, and secretes a variety of antibacterial materials, there is natural anti-corrosion effect, growth cycle is short, safe and harmless.Mulberries fermentation magma is applied to cosmetics, can effectively remove free radical, anti-oxidant, delaying cell aging, it is non-stimulated to skin safe.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of mulberries fermentation magma and its preparation method and application.
Background technique
Mulberries also known as sorosis, mulberry jujube, mulberry crow, are the general designations of Moraceae Mulberry plant mature fruit cluster.Mulberries are in aubergine or purple
Black, taste is fragrant and sweet, full of nutrition, is the food of dietotherapeutic.Mulberries contain various plants active constituent, great medicinal valence
Value.In the Tang Dynasty, the record that just there are mulberries to be used as medicine.With the development of science and technology researchers have confirmed that the medicinal valence of mulberries
Value.Containing functional active components such as mulberries polysaccharide, resveratrol, anthocyanin, flavonoids in mulberries, there is reducing blood lipid, drop blood
Sugar, removing free radical, oxidation resistant function and the growth to inhibition cancer cell, antibacterial anti-inflammatory, prevention of arterial atherosis have one
Constant current modulation effect.
Polysaccharide, anthocyanin, flavones, VE in mulberries etc., which all have, removes free radical, oxidation resistant function, therefore prepares
The related cosmetics of mulberries have broad mass market prospect.In natural plant raw material, the phenolic compounds such as polyphenol, flavones
Hydroxyl often is formed with sugar and glucosides substance, is existed in the form of conjugate, bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens these conjugates of beta-glucosidase enzyme hydrolysis) can be synthesized, more free polyphenol and flavones are discharged
Substance, to improve the antioxidant activity of plant material.Meanwhile can to secrete lipopeptid etc. a variety of by B.amyloliquefaciens
Antibacterial material has natural anti-corrosion effect.The invention is intended to provide one by B.amyloliquefaciens fermentation mulberries
Kind mulberries proferment pulp cosmetic and preparation method thereof.
Summary of the invention
In order to overcome the above-mentioned deficiency of the prior art, the present invention provides a kind of mulberries fermentation magma and preparation method thereof
And mulberries polyphenol, Huang in mulberries magma can be improved for bacillus amyloliquefaciens LX-12 in application, the fermenting microbe that the present invention uses
The content of the substances such as ketone, and a variety of antibacterial materials are secreted, there is natural anti-corrosion effect, growth cycle is short, safe and harmless；It will
Mulberries fermentation magma is applied to prepare cosmetics, and cosmetics can remove free radical, anti-oxidant, delaying cell aging.
The first purpose of this invention is to propose a kind of preparation method of mulberries fermentation magma, include the following steps: by
Mulberries, water and zymocyte liquid are mixed to get initial system, carry out fermented and cultured to the initial system, obtain mulberries proferment
Slurry；In the zymocyte liquid, strain is bacillus amyloliquefaciens LX-12 (Bacillus amyloliquefaciens LX-
12), deposit number CCTCC NO:M 2015234.
Further, the initial system is specific the preparation method comprises the following steps: mulberries and water are mixed with beating into juice, centrifugation point
From rear, filtering；It collects filtrate to sterilize to obtain mulberries fermentation substrate, then zymocyte liquid is inoculated in mulberries fermentation substrate.
Further, the zymocyte liquid concentration is 3.5 × 108-5×108CFU/mL, the zymocyte liquid is in mulberries
Inoculum concentration in fermentation substrate is 2%-5% (v/v).
Further, in the initial system, the mass ratio of mulberries and water is 1:(1.5-3).
Further, the condition of the fermented and cultured are as follows: fermented and cultured temperature is 30-42 DEG C, and fermented incubation time is
72-96h。
Further, the preparation method further include to after the fermented and cultured gained fermentation liquid successively sterilized, from
The heart, take supernatant obtain mulberries fermentation magma the step of.
Further, the condition of the sterilizing are as follows: sterilising temp is 110-125 DEG C, and sterilization time is 17-35 min；
The condition of the centrifugation are as follows: centrifugal rotational speed 4500-6700r/min, centrifugation time 18-45min.
Second object of the present invention is the mulberries fermentation for proposing that preparation method described in any of the above embodiments is prepared
Magma.
Third object of the present invention is to propose above-mentioned mulberries fermentation magma directly as with following at least one
Application in the cosmetics of kind function:
(1) anti-oxidant；
(2) free radical is removed；
(3) anti-aging.
Fourth object of the present invention is to propose that above-mentioned mulberries fermentation magma has following at least one function in preparation
Application in the cosmetics of energy:
(1) anti-oxidant；
(2) free radical is removed；
(3) anti-aging.
Compared with prior art, the beneficial effects of the present invention are:
(1) present invention uses microbe fermentation method, and preparation method is easy, remains mulberries polysaccharide, resveratrol, pattern
The whole components such as glycosides and its activity.
(2) it is more that mulberries in mulberries magma can be improved for bacillus amyloliquefaciens LX-12 in the fermenting microbe that the present invention uses
The content of the substances such as phenol, flavones, and a variety of antibacterial materials are secreted, there is natural anti-corrosion effect, growth cycle is short, safe nothing
Evil.
(3) present invention does not add any chemical component, pure natural production using bacillus amyloliquefaciens fermentation mulberries magma
Product, non-stimulated to skin when applied to cosmetics, skin is free of a burden.
(4) mulberries fermentation magma provided by the invention can remove free radical, anti-oxidant, delaying cell aging.
Specific embodiment
It shows that example illustrates certain embodiments of the present invention, and should not be construed as limiting model of the invention
It encloses.Present disclosure can be improved from material, method and reaction condition simultaneously, all these improvement should all
It falls within spirit and scope of the invention.
Quantitative experiment in following embodiment, is respectively provided with three repeated experiments, and results are averaged.
Mulberries structure used in the present invention is from Hubei Wuhan fruit supermarket.
Bacillus amyloliquefaciens LX-12 used in the present invention (Bacillus amyloliquefaciens LX-12), in
It is deposited in China typical culture collection center on April 15th, 2015, address is deposit number in Wuhan, China Wuhan University
CCTCC NO:M 2015234.DPPH structure is from sigma-Aldrich company, the U.S..
LB culture medium: peptone 10g/L, yeast powder 5g/L, sodium chloride 10g/L
Embodiment 1:
1, bacillus bacterium solution is prepared
1) the strain bacillus amyloliquefaciens LX-12 of freezing is inoculated into LB solid medium tablets, 37 DEG C of trainings
12h is supported, its activation is made；
2) by the strain transfer of activation into LB liquid seed culture medium, 37 DEG C, 180rpm cultivates 8-14 h, and brood cell is made
Bacillus bacterium solution, bacillus LX-12 bacterial concentration are 3.5 × 108CFU/mL。
2, mulberries fermentation magma is prepared
1) 25g mulberries and 50mL water are mixed with beating into juice, and 4000rpm/min is centrifuged 10min, then remaining with filtered through gauze
Pomace carries out pasteurization.
2) zymocyte liquid for preparing step 1 is inoculated in mulberries fermentation substrate according to the inoculum concentration of 2% (v/v), and 37
DEG C, 180rpm cultivates 96h.Fermentation system, as tunning are collected after fermentation.
3) tunning for taking step 2) to obtain, 115-121 DEG C of sterilizing 15-25min.
4) product of step 3) is taken, 10000rpm is centrifuged 20min, collects supernatant, as mulberries fermentation magma.
Embodiment 2:
1, bacillus bacterium solution is prepared
The method of bacillus amyloliquefaciens LX-12 bacterium solution is prepared with embodiment 1, bacillus LX-12 bacterial concentration is
4.5×108CFU/mL。
2, mulberries fermentation magma is prepared
1) 20g mulberries and 30mL water are mixed with beating into juice, and 4000rpm/min is centrifuged 10min, then remaining with filtered through gauze
Pomace carries out pasteurization.
2) zymocyte liquid for preparing step 1 is inoculated in mulberries fermentation substrate according to the inoculum concentration of 3.5% (v/v)
In, 30 DEG C, 180rpm cultivates 84h.Fermentation system, as tunning are collected after fermentation.
3) tunning for taking step 2) to obtain, 115-121 DEG C of sterilizing 15-25min.
4) product of step 3) is taken, 8000rpm is centrifuged 30min, collects supernatant, as mulberries fermentation magma.
Embodiment 3:
1, bacillus bacterium solution is prepared
The method of bacillus amyloliquefaciens LX-12 bacterium solution is prepared with embodiment 1, bacillus LX-12 bacterial concentration is 5
×108CFU/mL。
2, mulberries fermentation magma is prepared
1) 30g mulberries and 90mL water are mixed with beating into juice, and 4000rpm/min is centrifuged 10min, then remaining with filtered through gauze
Pomace carries out pasteurization.
2) zymocyte liquid for preparing step 1 is inoculated in mulberries fermentation substrate according to the inoculum concentration of 5% (v/v),
42 DEG C, 180rpm cultivates 72h.Fermentation system, as tunning are collected after fermentation.
3) tunning for taking step 2) to obtain, 115-121 DEG C of sterilizing 15-25min.
4) product of step 3) is taken, 12000rpm is centrifuged 15min, collects supernatant, as mulberries fermentation magma.
Embodiment 4: application of the mulberries fermentation magma in cosmetics
One, the property of mulberries fermentation magma
By resulting fermentation magma prepared by embodiment 1-3 through active carbon decoloring, de-taste.Its appearance be thick liquid,
Color is faint yellow to brown color.PH value 6.1-6.8, viscosity 190-230cP, soluble solids content 3.0-3.5%, bacterium
It falls sum and is less than 50CFU/mL, no pathogenic bacteria detection.According to hygienic standards for cosmetics GB7916-87, cosmetics total number of bacteria is not
Higher than 1000CFU/mL, so this fermentation magma meets cosmetics quality requirement.
Two, mulberries fermentation magma constituent analysis
Analysis of antioxidant ingredients is carried out to the mulberries fermentation magma of embodiment 1-3 preparation, wherein fermentation magma is added 10 times
The methanol (m/v) of volume 80% is diluted, and 0.2mL dilute sample is added in the forint phenol reagent (1N) of 0.2mL, reaction
0.4mL Na is added in 3min2CO3Solution (1N) reacts at room temperature 90min, and 2mL deionized water is added, and 725nm measures absorbance,
Standard curve is made using gallic acid as standard items (10,20,30,40,50 μ g/mL), total phenol content is expressed equivalently as mg and does not eat
Sub- acid/mL fermentation magma.The measurement of general flavone content is using rutin as standard items (5,10,15,20,25 μ g/mL), dilution fermentation
Magma sample, detects 258nm absorbance, and general flavone content is expressed equivalently as mg rutin/mL fermentation magma.Acquired results such as table
1:
Mulberries fermentation magma constituent analysis in 1 embodiment 1-3 of table
| General flavone (mg rutin/mL) | Total phenol (mg gallic acid/mL) | |
| Embodiment 1 | 1729.07 | 510.17 |
| Embodiment 2 | 1778.49 | 546.80 |
| Embodiment 3 | 1795.93 | 571.22 |
As shown in Table 1, in mulberries magma prepared by the fermenting microbe bacillus amyloliquefaciens LX-12 that the present invention uses
Rich in substances such as polyphenol, flavones.
Three, mulberries fermentation magma anti-oxidation efficacy analysis
DPPH is a kind of general organic free radical, the oxidation resistance of assessment antioxidant is often used to, organic
It is more stable in solvent, it is purple, and have special absorption peak at 517nm, and when encountering free radical scavenger, the orphan of DPPH
Electronics is paired and makes its colour fading, the peak value of its maximum absorption wavelength is caused to become smaller.Therefore, measurement light absorption value can be passed through
Reduction degree evaluates sample to the elimination effect of DPPH free radical, reflects the antioxidant activity of sample.
The step of DPPH radicals scavenging is tested:
1, the mulberries fermentation magma of Example 1-3 is prepared into 10% fermentation magma sample respectively with methanol mixed diluting
Product, for several times, 10000rpm is centrifuged 5min for oscillation.
2, each 500 μ L of 3 kinds of centrifugates in step 1 is taken, is mixed respectively with 75 μm of ol/L DPPH- methanol solutions of 2.5mL,
Room temperature is protected from light 90min.Control group is that 500 μ L methanol are mixed with 75 μm of ol/L DPPH- methanol solutions of 2.5mL, and room temperature is kept away
Light reaction 90min.
After reaction, absorbance is detected under 517nm.
Clearance rate (%)=(1- sample absorbance/control absorbance) × 100%
Mulberries fermentation magma removes DPPH free radical effect such as table 2.
The mulberries fermentation magma of table 2 10% is to DPPH free radical scavenging activity
| DPPH clearance rate | |
| Embodiment 1 | 90% |
| Embodiment 2 | 92% |
| Embodiment 3 | 94% |
As shown in Table 2, the mulberries fermentation magma prepared in embodiment 1-3 has very strong removing energy to DPPH free radical
Power.10% mulberries fermentation magma has all reached 90% or more to DPPH free radical scavenging activity.Illustrate that mulberries fermentation magma has
The very strong ability for removing free radical, promotes cell metabolism, enhances cell viability, improves body mechanism and function, improves body
Vitality plays the effect of its anti-aging to delay cell senescence.
Four, mulberries fermentation magma property evaluation safe to the human body
Cosmetics patch test is mainly used for detecting the irritation of cosmetic product.The present invention passes through human closed formula
Patch test evaluates the safety of mulberries fermentation magma, assesses its potential skin irritation.
Subjects: 30 volunteers of the range of age at 18-60 years old
Test method: test group: 0.2mL mulberries ferment magma dropwise addition in filter paper, place into spot tester.Blank pair
According to group: 0.2mL distilled water is added dropwise in another filter paper, is put into spot tester.
Test position is that spot tester is fixed on the outside of volunteer's upper arm with non-stimulated adhesive tape on the outside of upper arm.When test
Between for is for 24 hours for the accuracy of test result, dduring test, volunteer cannot remove spot tester, test department as requested
Position can not contact water.
It removes spot tester afterwards for 24 hours, stands 30min, disappear to recipient site impression, observe the reaction of skin.If test is
Feminine gender, also need observation for 24 hours, after 48h, the reaction of skin.
Patch test results are as shown in table 3.The meaning that each symbol indicates in table 3 is as follows:
"-" indicates negative reaction；
" ± ", indicates suspicious reaction: only faint erythema；
"+" indicates weakly positive reaction (erythematous response): erythema, oedema, can have papule at infiltration；
" ++ " indicates strong positive reaction (bleb reaction): erythema, infiltration, oedema, papule, bleb；Reaction can exceed tested
Area；
" +++ " indicates pole positive reaction (reaction of amalgamation bleb): obvious erythema, severe infiltration, oedema, amalgamation blister
Rash；Reaction exceeds tested area.
3 patch test results of table
The mulberries fermentation magma that embodiment 1-3 is obtained as can be seen from Table 3 does not generate suspicious reaction, illustrates preparation of the present invention
Mulberries proferment pulp cosmetic have safety, adverse reaction will not be brought to human body.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited to
This, anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention
And its inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of preparation method of mulberries fermentation magma, which comprises the steps of: by mulberries, water and zymocyte liquid
It is mixed to get initial system, fermented and cultured is carried out to the initial system, obtains mulberries fermentation magma；In the zymocyte liquid,
Strain be bacillus amyloliquefaciens LX-12 (Bacillus amyloliquefaciens LX-12), deposit number CCTCC NO:
M 2015234。
2. a kind of preparation method of mulberries fermentation magma according to claim 1, which is characterized in that the initial system
Specifically the preparation method comprises the following steps: mulberries and water are mixed with beating into juice, after centrifuge separation, filtering；Filtrate is collected to sterilize to obtain mulberries hair
Ferment substrate, then zymocyte liquid is inoculated in mulberries fermentation substrate.
3. a kind of preparation method of mulberries fermentation magma according to claim 2, which is characterized in that the zymocyte liquid is dense
Degree is 3.5 × 108-5×108CFU/mL, inoculum concentration of the zymocyte liquid in mulberries fermentation substrate are 2%-5% (v/v).
4. according to claim 1 or a kind of preparation methods of 2 described in any item mulberries fermentation magma, which is characterized in that described
In initial system, the mass ratio of mulberries and water is 1:(1.5-3).
5. a kind of preparation method of mulberries fermentation magma according to claim 1, which is characterized in that the fermented and cultured
Condition are as follows: fermented and cultured temperature is 30-42 DEG C, fermented incubation time 72-96h.
6. a kind of preparation method of mulberries fermentation magma according to claim 1, which is characterized in that the preparation method is also
Including successively being sterilized, being centrifuged to gained fermentation liquid after the fermented and cultured, supernatant is taken to obtain the step of mulberries fermentation magma
Suddenly.
7. a kind of preparation method of mulberries fermentation magma according to claim 6, which is characterized in that the condition of the sterilizing
Are as follows: sterilising temp is 115-121 DEG C, sterilization time 15-25min；The condition of the centrifugation are as follows: centrifugal rotational speed 8000-
12000r/min, centrifugation time 15-30min.
8. the mulberries fermentation magma being prepared such as the described in any item preparation methods of claim 1-7.
9. mulberries fermentation magma according to any one of claims 8 is directly as answering in the cosmetics with following at least one functions
With:
(1) anti-oxidant；
(2) free radical is removed；
(3) anti-aging.
10. application of the mulberries fermentation magma according to any one of claims 8 in the cosmetics that preparation has following at least one functions:
(1) anti-oxidant；
(2) free radical is removed；
(3) anti-aging.
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Cited By (4)
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| CN113398056A (en) * | 2021-07-26 | 2021-09-17 | 上海梵中生物科技有限公司 | Biological plant extract skin beautifying mask cream and preparation method thereof |
| CN115154363A (en) * | 2022-09-01 | 2022-10-11 | 广东省农业科学院蚕业与农产品加工研究所 | Mulberry resource oligosaccharide moisturizing composition and preparation method and application thereof |
| CN115337335A (en) * | 2022-08-19 | 2022-11-15 | 广州市科贝思生物技术有限公司 | Preparation method and application of mulberry extract with anti-saccharification effect |
| CN115400054A (en) * | 2022-09-01 | 2022-11-29 | 广东省农业科学院蚕业与农产品加工研究所 | Preparation method and application of whitening and antioxidant composition based on mulberry resources |
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Application publication date: 20190628 |