CN111514077A - Lactobacillus pentosus aloe fermented raw juice, preparation method, preservation method and application thereof - Google Patents
Lactobacillus pentosus aloe fermented raw juice, preparation method, preservation method and application thereof Download PDFInfo
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- CN111514077A CN111514077A CN202010253192.2A CN202010253192A CN111514077A CN 111514077 A CN111514077 A CN 111514077A CN 202010253192 A CN202010253192 A CN 202010253192A CN 111514077 A CN111514077 A CN 111514077A
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- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000009959 type I hypersensitivity Effects 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
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Abstract
The invention discloses lactobacillus pentosus aloe fermentation raw stock, a preparation method, a preservation method and application thereof, wherein the preparation method comprises the following steps: s1, processing raw materials; s2, domesticating zymocyte; s3, preparation of fermentation raw pulp: preparing the aloe juice prepared in the step S1 into an aloe fermentation culture medium, inoculating 1-10% by volume of the fermentation seed liquid treated in the step S2 into the aloe fermentation culture medium, and standing and culturing at 30-38 ℃ for 1-20 days; s4, centrifuging to obtain supernatant, and sterilizing to obtain the fermented raw stock. Compared with the prior art, the oxidation resistance, the whitening effect and the like of the fermentation raw stock prepared by the scheme are greatly improved, the lactobacillus pentosus is a food safe strain and is safe and harmless, all the added materials are safe and nontoxic reagents, the safety is high, no pollution is caused, the stability is good, the preparation process is mild, the operation is simple and convenient, and the industrial application prospect is good.
Description
Technical Field
The invention relates to the technical field of biological fermentation engineering, in particular to lactobacillus pentosus aloe fermentation raw stock, and a preparation method, a preservation method and application thereof.
Background
Aloe is a perennial evergreen herbaceous plant, and is rich in a plurality of active ingredients such as anthraquinone substances, saccharides, amino acids, organic acids, mineral substances and the like, so that the aloe has a plurality of biological activities such as oxidation resistance, aging resistance, inflammation diminishing, bacteria resistance, cancer resistance, healing promotion and the like, and is widely applied to the fields of food, beauty, health care, medicine and the like. How to effectively extract the active ingredients in the aloe vera extract has important significance for improving the industrial application value of the aloe vera.
The traditional extraction method mainly comprises a solvent extraction method and an ultrasonic extraction method, wherein the two methods have the defects of low yield, low purity, serious pollution and difficult industrial production of the extracted aloe active substances, and particularly in the solvent extraction method, high-temperature conditions are required to be combined with chemical solvent extraction, and certain influence (such as weakening of the anti-inflammatory, antioxidant and whitening effects of the product) is caused on the aloe active ingredients. In recent years, the extraction of active ingredients in plants by using a microbial fermentation technology has become a trend, and the technology has unique advantages in the aspect of extraction of plant active substances due to the advantages of strong reaction specificity, few byproducts, mild reaction conditions, cleanness, environmental protection and the like. The Chinese invention patent application CN105853303A discloses a preparation method of a probiotic Lucai mask, and specifically discloses that six active probiotics (specifically bifidobacterium longum, bifidobacterium breve, bifidobacterium adolescentis, lactobacillus rhamnosus, lactobacillus plantarum and lactobacillus delbrueckii) are utilized for fermentation, compared with the traditional water extraction method, the method can retain active ingredients in plants to a greater extent, but the DPPH clearance rate is only 29.92%. Chinese invention patent CN105250169A discloses an aloe fermented raw stock cosmetic and a preparation method and application thereof, and particularly discloses that streptococcus thermophilus is used for fermenting aloe raw stock, and the method can also reserve active ingredients in plants to the maximum extent and has a certain whitening effect, but the method still cannot meet the increasing whitening requirements of people. Therefore, the development of a new fermentation technology is of great significance for improving the industrial application value of the active ingredients in the aloe.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides a preparation method of the lactobacillus pentosus aloe fermentation raw stock, and the fermentation raw stock prepared by the method has good biological activity.
The invention also provides the fermented raw pulp prepared by the method.
The invention also provides application of the fermentation raw stock.
The invention also provides a preservation method of the fermentation raw stock.
The preparation method according to the embodiment of the first aspect of the invention comprises the following steps:
s1, raw material treatment: squeezing sterilized aloe to obtain juice;
s2, domestication of zymocyte: preparing the aloe juice treated in the step S1 into an aloe seed culture medium, culturing the activated 2-3 generations of lactobacillus pentosus at 30-38 ℃ for 16-24 h, inoculating the lactobacillus pentosus into the aloe seed culture medium according to the volume ratio of 1-3%, and continuously culturing at 30-38 ℃ for 16-24 h to obtain a fermented seed solution for later use;
s3, preparation of fermentation raw pulp: preparing the aloe juice prepared in the step S1 into an aloe fermentation culture medium, inoculating 1-10% of the fermentation seed liquid treated in the step S2 into the aloe fermentation culture medium according to the volume ratio, and standing and culturing for 1-20 days (preferably 1-10 days; more preferably 2-5 days) at 30-38 ℃;
s4, centrifuging to obtain supernatant, and sterilizing to obtain the fermented raw stock.
The preparation method provided by the embodiment of the invention has at least the following beneficial effects: fermenting aloe with domesticated Lactobacillus pentosus, and utilizing microbial enzymolysis technology to loosen part of aloe fiber, effectively release active ingredients, and metabolize to generate new active substances to improve the efficacy of the extract; the prepared lactobacillus pentosus aloe fermentation raw stock not only has good anti-inflammatory effect and whitening effect of aloe, but also has the characteristic of excellent oxidation resistance of fermentation liquor obtained by synergy of lactobacillus pentosus fermentation metabolites; meanwhile, the aloe flesh juice is used as the seed liquid, and compared with MRS broth used as the seed liquid, the obtained fermentation raw pulp is light yellow, and has light fermentation smell and proper smell; compared with the prior art, the antioxidant and whitening effects of the fermented raw stock prepared by the scheme of the invention are greatly improved, in addition, the lactobacillus pentosus used by the scheme of the invention is a safe strain for food, is safe and harmless, all the added materials are safe and nontoxic reagents, the safety is high, no pollution is caused, the stability is good, the prepared fermented raw stock can be directly used for preparing cosmetics, the preparation process is mild, the operation is simple and convenient, and the industrial application prospect is good.
According to some embodiments of the invention, the aloe is further pre-treated prior to sterilization by: peeling Aloe, collecting flesh, washing with water, and adding Vitamin C (VC) for protecting color.
According to some embodiments of the invention, the aloe is pasteurized. Performing pasteurization (water bath at 80 deg.C for 30 min) to prevent loss of active components at high temperature.
According to some embodiments of the invention, the aloe vera is 2-3 years old. The aloe of this kind of age has more polyphenol flavone active components.
According to some embodiments of the invention, the number of washing operations is 1 to 5; preferably 3 times.
According to some embodiments of the invention, the VC is added in a mass ratio of 0.01-0.2%.
According to some embodiments of the invention, the aloe seed medium is prepared by the steps of: adding the aloe juice into a sterilized solution containing a carbon source and an exogenous nitrogen source in a mass ratio of more than 60%, uniformly mixing, and adjusting the pH value to 6.5-7.0 (preferably 6.8).
According to some embodiments of the present invention, the step S2 is to culture the aloe seeds inoculated with Lactobacillus pentosus for 16-24 h at 30-38 ℃ with shaking.
According to some embodiments of the invention, the aloe juice is added in a proportion of 60-100%; preferably 60 to 80%.
According to some embodiments of the invention, the carbon source is glucose; preferably, the mass percentage of the glucose is 1-4%.
According to some embodiments of the invention, the exogenous nitrogen source is selected from at least one of yeast extract powder or beef extract powder.
According to some embodiments of the present invention, the aloe fermentation medium of step S3 is prepared by: mixing aloe juice with sterilized sucrose-containing solution, and mixing; preferably, the mass ratio of the aloe juice in the aloe fermentation medium is 30-60%; preferably, the mass volume of the sucrose in the sucrose-containing solution accounts for 3-7%.
According to some embodiments of the invention, in the step S4, the centrifugal speed is 3000 to 10000 rpm; the time is 10-30 min.
According to some embodiments of the present invention, the sterilization in step S4 is performed by filter sterilization; preferably, the fermentation raw pulp is taken and filtered to enable the fermentation raw pulp to pass through 0.22 mu m. And the filtration sterilization mode is adopted, so that the failure of active ingredients in the primary fermentation broth due to high-temperature sterilization operation is avoided.
The fermentation raw stock according to the second aspect of the embodiment of the invention is prepared by the method.
The fermentation raw stock provided by the embodiment of the invention at least has the following beneficial effects: the odor of the fermentation raw stock is effectively improved, the color of the fermentation liquor is reduced, the fermentation effect of the pentobacterium is improved, the fermentation raw stock can be directly used for developing products such as cosmetics, and the like, and all materials added in the preparation process are safe and non-toxic reagents, so that the safety and the pollution are high, and the stability is good; the fermentation broth raw stock has good anti-allergy effect and whitening effect of aloe, and has excellent oxidation resistance of lactobacillus pentosus metabolites.
According to the application of the embodiment of the third aspect of the invention, the fermented protoplasm is applied to the preparation of cosmetics.
According to the application of the third aspect of the embodiment of the invention, the fermented raw pulp is applied to the preparation of an antioxidant, an anti-allergy agent or a whitening agent.
The application of the embodiment of the invention has at least the following beneficial effects: the fermentation raw stock has good anti-inflammatory, antioxidant, whitening and other effects, the addition amount is preferably 1-10%, the fermentation raw stock can have good effects within the range, the effects are gradually increased along with the gradual increase of the concentration of a sample, and the fermentation raw stock has good application prospects in the field of cosmetics.
A saving method according to an embodiment of a fourth aspect of the present invention includes the steps of: adding polyalcohol into the fermented primary pulp.
According to some embodiments of the invention, the polyol comprises hexylene glycol.
According to some embodiments of the invention, the preservation method comprises the steps of: adding p-hydroxyacetophenone with the mass volume ratio of not more than 1%, hexanediol with the volume ratio of not more than 1% and ethylhexyl glycerol with the volume ratio of not more than 1% into the fermentation raw stock.
According to some embodiments of the invention, the ratio of p-hydroxyacetophenone to hexylene glycol is 0.5% by volume, the ratio of hexylene glycol to hexyl.
The preservation method provided by the embodiment of the invention has at least the following beneficial effects: the fermentation raw stock prepared by the scheme of the invention can have good anticorrosion effect by mixing with the polyhydric alcohol with anticorrosion effect; the preservative effect is evaluated by a CTFA recommended method, and the result shows that the preservative effect is excellent, which indicates that the preservative system has extremely strong inhibition and killing effects on microorganisms and can ensure that the aloe fermentation raw stock is not easily polluted by the microorganisms during storage and use.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
FIG. 1 is a colony morphology and gram stain pattern (10 x 100) of Lactobacillus pentosus in example 1 of the present invention;
FIG. 2 shows the hyaluronidase inhibition effect of aloe vera fermented puree measured in example 1 of the present invention;
FIG. 3 shows the inhibition effect of aloe fermented puree on tyrosinase, measured in example 1 of the present invention;
FIG. 4 is a graph showing the effect of aloe fermentation broth on scavenging ABTS +. free radicals measured in example 1 of the present invention;
FIG. 5 is a graph showing the antioxidant properties of aloe fermentation broth prepared by different strains according to comparative example 1;
FIG. 6 is a graph showing the effect of different nitrogen source concentrations on the growth of Lactobacillus pentosus in comparative example 2 of the present invention.
Detailed Description
In order to explain technical contents, achieved objects, and effects of the present invention in detail, the following description is made with reference to the accompanying drawings in combination with the embodiments.
The first embodiment of the invention is as follows: a lactobacillus pentosus aloe fermentation raw stock and a preparation method and application thereof are disclosed, wherein the preparation method comprises the following steps:
(1) processing the aloe raw material: cleaning fresh and full-fleshed aloe, peeling, collecting flesh, washing with water for 3 times, cutting into pieces, adding 0.02% VC, protecting color, packaging into a sealed bag, and sealing. Performing sterilization treatment by adopting a pasteurization method: water bath at 80 deg.C for 30min, cooling to room temperature, and storing in 4 deg.C refrigerator.
(2) Domestication of lactobacillus pentosus: placing sterilized aloe pulp in a juicer, squeezing, adding into sterilized solution containing glucose (3%, m/v) and yeast extract powder (0.6%, m/v) at a ratio of 80%, adjusting pH to 6.8, and mixing to obtain aloe seed culture medium.
Lactobacillus pentosus (commercially available, and named Lactobacillus plantarum, CICC24202) with 1-ring activated 3 generations is inoculated into MRS broth and cultured in a shaker at 37 deg.C and 220r/min for 24 h. 2% of the inoculum size was inoculated into aloe seed medium, cultured for 18 hours at 37 ℃ in a shaker at 220r/min, the number of colonies of lactobacillus pentosus was counted by plate counting, and the growth state of lactobacillus pentosus was observed by microscopic examination, as shown in fig. 1. As can be seen from FIG. 1, the colonies are tiny round white colonies, convex and have regular edges; the cell is rod-shaped, short chain-shaped and non-spore, and belongs to gram-positive bacteria.
(3) Preparing aloe fermented raw stock: inoculating the aloe seed culture medium at 10% (V/V) into aloe fermentation culture medium containing aloe pulp 40% (V/V) and sucrose 7% (m/V), and standing and culturing at 37 deg.C for 72 hr. After fermentation, the aloe fermentation liquor is put in a centrifuge for 30min at 10000 r/min, and supernatant is taken. Filtering to remove bacteria, filtering with 0.22 μm filter membrane, and collecting filtrate in a clean sterilized glass container.
The fermentation raw stock prepared by the method is used for testing the anti-inflammatory, antioxidant and whitening effects of the fermentation raw stock to verify the application prospect of the fermentation raw stock in the preparation of anti-inflammatory, antioxidant and whitening agents, and the specific process is as follows:
1) anti-allergic effect test of aloe fermented puree (Hyaluronidase inhibitory effect)
Hyaluronic acid is a component in a tissue matrix for limiting the diffusion of water and other extracellular substances, and after being hydrolyzed by hyaluronidase, cells become non-viscous, so that new media for degranulation and synthesis of the cells are exuded, a biological effect is exerted, and an immediate type anaphylactic reaction is caused. Hyaluronidase has a strong correlation with allergy and is therefore commonly used in the degree of inhibition of hyaluronidase by a subject to reflect its alleviating and ameliorating effects on type i hypersensitivity.
Taking 0.1mL of CaCl2Reacting the solution (0.25mmol/L) with 0.5mL of hyaluronidase solution in water bath at 37 ℃ for 20 min; adding 0.5mL of fermentation raw stock solution sample, and keeping the temperature for 20 min; then adding 0.5mL of sodium hyaluronate solution (0.5g/L), reacting for 30min, taking out and cooling for 5 min; adding 0.1mL of sodium hydroxide (0.4mol/L) and 0.5mL of acetylacetone solution (3.5mL of acetylacetone dissolved in 50mL of 1.0mol/L sodium carbonate solution), boiling in water bath for 15min, and immediately transferring to ice water bath for 5 min; 1mL of Ellisib reagent was added dropwise, diluted with 3mL of absolute ethanol, left at room temperature for 20min for color development, and the absorbance was measured at 530 nm.
The hyaluronidase inhibition (%) was calculated according to the following formula:
D(％)＝[(A-B)-(C-D)]/(A-B)×100％
wherein A is the absorbance of the control solution (the sample solution is replaced by acetic acid buffer solution); b is the absorbance of the control blank solution (the sample solution and the enzyme solution are replaced by acetic acid buffer solution); c is the absorbance of the sample solution; d is the absorbance of the sample blank solution (the enzyme solution is replaced by acetic acid buffer solution).
In the test process, samples of fermentation raw stock solutions with different volume concentrations are taken to carry out parallel experiments, meanwhile, 31mg/ml dipotassium glycyrrhizinate solution is taken as a control experiment, and partial test results are shown in figure 2.
As can be seen from FIG. 2, aloe vera fermented puree has excellent inhibitory effect on hyaluronidase in the study of anti-allergy effect of aloe vera fermented puree. When the volume concentration of the aloe fermented raw stock is within the range of 1-5%, the inhibition rate of the aloe fermented raw stock on hyaluronidase is gradually increased along with the gradual increase of the sample concentration. When the concentration of the sample is 5%, the inhibition rate of the hyaluronidase reaches 53.25%. In addition, the half inhibitory concentration IC of the aloe fermented puree on hyaluronidase is also measured504.8 percent; therefore, the aloe fermented raw stock prepared by the embodiment of the invention has good relieving and improving effects on anaphylactic reaction and inflammation, and has wide application prospect in development and application of anti-inflammatory skin care products.
2) Whitening effect test of Aloe fermentation stock (Mushroom tyrosinase inhibitory effect)
Tyrosinase is the rate-limiting enzyme in the melanin synthesis pathway, and mainly affects the synthesis of melanin by reacting with tyrosine to oxidize the tyrosine into dopa acid and further oxidizing the dopa acid into dopaquinone. In the in vitro test, mushroom tyrosinase is usually selected for testing to determine the tyrosinase inhibition rate of the whitening agent, and the whitening effect is evaluated according to the tyrosinase inhibition rate.
Using 6mmol/L L-tyrosine (L-tyrosine, L-TRY) as a substrate, each solution was added in the following Table 1 in a 4mL test system at 0.05mmol/L (pH 6.8).
TABLE 1 tyrosinase inhibition assay System
Remarking: and adding the reaction liquid in sequence, and entering the next step after the corresponding treatment is finished, wherein the "-" represents no addition.
The tyrosinase inhibition rate of the sample was calculated according to the formula:
D(％)＝[(A3-A4)-(A1-A2)]/(A3-A4)×100％
in the test process, fermentation raw stock solution samples with different volume concentrations are taken for parallel experiments, meanwhile, 280 mu g/ml arbutin solution is taken as a control experiment, and partial test results are shown in figure 3.
The test results are shown in FIG. 3, and it can be seen from FIG. 3 that the whitening effect on aloe vera fermented puree isOn the research, the aloe fermentation raw stock has good inhibition effect on mushroom tyrosinase. At 6 vol%, the tyrosinase inhibition rate of aloe fermented raw juice was 52.66% (from which the IC was estimated to be50<6.0%). In addition, when the volume concentration is 10%, the tyrosinase inhibition rate is 64.55%, and the effect is equivalent to that of arbutin (280 μ g/mL) serving as a control whitening agent (the tyrosinase inhibition rate is 63.32%). The aloe fermented raw stock has good capability of inhibiting tyrosinase activity in vitro, can inhibit the generation of melanin, and has certain whitening effect.
3) Antioxidant activity test of aloe fermentation raw stock (ABTS +. free radical scavenging action)
2,2 '-biazoyl-bis-3-ethylbenzthiazoline-6-sulfonic acid (2, 2' -azino-bis (3-ethyllbenzothiazoline-6-sulfonic acid, ABTS) is oxidized by potassium persulfate to generate blue-green cationic free radical ABTS +. the maximum absorption peak exists at the wavelength of 734nm, when antioxidant exists, ABTS +. can react with the cationic free radical to generate colorless ABTS, and the change of the absorption value at the wavelength of 734nm is measured by a microplate reader to reflect the ABTS +. the free radical scavenging activity of the antioxidant.
ABTS +. preparation of working solution: mixing 7mmoL/L ABTS + solution and 2.45mmoL/L potassium persulfate solution in equal volume, and standing overnight for 16h at room temperature in the dark to obtain ABTS + stock solution. The ABTS + stock solution was diluted with a phosphate buffer (10mmoL/L, pH 7.4) so that the absorbance at a wavelength of 734nm became 0.700. + -. 0.020 to obtain ABTS +. working solution.
TABLE 2ABTS +. free radical scavenging assay System
| A1 | A2 | A3 | A4 | |
| Deionized water | 50μL | 50μL | - | - |
| ABTS +. working solution | 150μL | - | 150μL | - |
| Sample to be tested | - | - | 50μL | 50μL |
| Phosphate buffer | - | 150μL | - | 150μL |
Remarking: the solutions were added in the volume shown in Table 2, mixed well, reacted at room temperature in the dark for 10min, and the absorbance was measured at 734 nm.
D (ABTS +. radical clearance,%) is [ (a)1-A2)-(A3-A4)]/(A1-A2)×100％
In the test process, fermentation raw stock solution samples with different volume concentrations are taken to carry out parallel experiments, and partial test results are shown in figure 4.
As can be seen from FIG. 4, in the research of the antioxidant activity of the aloe fermented raw juice, when the volume concentration of the aloe fermented raw juice is in the range of 1-10%, the scavenging capacity of the aloe fermented raw juice on ABTS +. free radicals is gradually increased along with the gradual increase of the sample concentration, and obvious dose dependence is presented. Half-clearing concentration IC of aloe fermentation raw stock on ABTS +. free radical502.0%. Therefore, the microbial fermentation is beneficial to the release of active ingredients of the aloe fermented protoplasm, and the aloe fermented protoplasm has better antioxidant capacity by combining the synergistic effect of the antioxidant metabolites of the lactobacillus plantarum, and has an important effect on long-acting antioxidation of skin.
In summary, the aloe fermented puree prepared by using lactobacillus pentosus according to the embodiments of the present invention has excellent hyaluronidase Inhibition (IC) effect504.8%), tyrosinase activity Inhibition (IC)50<6.0%) and strong ABTS +. free radical scavenging action (IC)502.0%), and has good safety. Therefore, the aloe fermentation broth raw stock is added into cosmetics according to the proportion of 2-10% for application, and has excellent anti-allergy effect (hyaluronidase inhibition rate), whitening effect (tyrosinase inhibition rate) and antioxidant effect (ABTS +. free radical scavenging effect).
The first comparative example of the invention is as follows: influence of different fermentation strains on antioxidant activity of aloe fermentation raw stock
The aloe product has antioxidant activity and can be widely applied to the fields of food, beauty treatment, health care, medicine and the like. Thus, two different control ferments were selected: the composite probiotics (bifidobacterium longum, lactobacillus acidophilus, lactobacillus casei, streptococcus thermophilus, bifidobacterium breve and bifidobacterium lactis ═ 1: 1: 1: 1: 1) and yeast (Angel) ferment the aloe under the same conditions as the lactobacillus pentosus in the embodiment, and the influence of the antioxidant effect of the aloe culture fermentation raw stock is researched by taking ABTS & lt + & gt free radical scavenging effect as an index (see the efficacy test of the aloe fermentation raw stock by an ABTS & lt + & gt free radical scavenging test method).
Meanwhile, the ABTS & lt + & gt free radical activity removing effect of the aloe liquid prepared by fermenting the aloe juice for 3 days in different fermentation modes (oscillating fermentation and standing fermentation) is also examined. The experimental results are shown in fig. 5, wherein the relative ABTS +. free radical scavenging activity of the fermented aloe broth after fermentation was calculated with the ABTS +. free radical scavenging activity of the unfermented aloe gravy being 100%.
As can be seen from FIG. 5, the supernatants obtained by shaking fermentation with complex bacteria and yeast and the aloe supernatant obtained by fermentation with Lactobacillus pentosus in a standing condition have better ABTS +. free radical scavenging activity, respectively, compared to the aloe gravy medium before fermentation. Among the three, the ability of eliminating ABTS + free radicals of the aloe fermentation liquid obtained by standing and fermenting the lactobacillus pentosus is improved most obviously.
The second comparative example of the present invention is: exploration of influence of different nitrogen source addition concentrations on fermentation effect
If MRS broth is used as a culture medium, filtrate obtained by fermentation is dark yellow, and the fermentation smell is heavy, so that the invention selects and uses the aloe gravy plant nutrient culture medium as a basis, and adds a proper nitrogen source (yeast extract powder) and a proper carbon source (3 percent of glucose, m/v) ratio to replace the MRS broth, and simultaneously slightly improves the aloe fermentation culture medium. The aloe plant culture medium is added with a proper amount of exogenous nitrogen source, which is beneficial to the growth of lactobacillus pentosus and has good growth condition. The concentration of the lactobacillus pentosus liquid reaches 108CFU/mL, the concentration of the strain liquid required for fermentation is reached, and the experimental result is shown in FIG. 6. In order to effectively improve the color and smell of the aloe fermentation broth, it is preferable that the exogenous nitrogen source is added at a concentration of 0.6%. The aloe fermentation raw stock finally obtained is transparent and light yellow, has lighter fermentation smell and proper smell, and is superior to the fermentation effect of directly taking MRS broth as seed liquid. And simultaneously observing the growth state of the lactobacillus pentosus by microscopic examination.
The second embodiment of the invention is as follows: a preservation method of pentose lactic bacteria aloe fermentation protoplasm comprises the following steps: taking the aloe fermented raw stock prepared in the first embodiment, and adding 0.5% (m/v) of p-hydroxyacetophenone, 0.5% (v/v) of hexanediol and 0.05% (v/v) of ethylhexyl glycerol.
The preservative effect of the preservative system on the aloe fermentation raw stock in the normal temperature storage process is evaluated by referring to a preservative efficacy evaluation method (namely a preservative challenge test) recommended by American Cosmetic, Toiletry and perfume Association (CTFA). The method specifically comprises the following steps: respectively adding mixed bacterial suspension (Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa) and mixed fungal suspension (Aspergillus niger and Candida albicans) into a sample to be tested by one-step bacteria adding method, wherein the concentration of the mixed bacterial suspension is 105～106CFU/mL, mixed fungal suspension concentration of 104～105CFU/m L, cultured in an incubator at 28 ℃. At 7, 14, 21 and 28 days, sampling, coating and counting to analyze the colony number, evaluating the preservative effect of the preservative system according to the percentage reduction of the colony number, and testing results are shown in the following table 3:
TABLE 3 test results of preservative system for aloe fermentation puree
The third comparative example of the invention is: a preservation method of pentose lactic bacteria aloe fermentation protoplasm comprises the following steps: the aloe fermented puree prepared in example one was added with 0.2% (w/v) lauroyl arginine ethyl ester HCl, 1.0% (v/v) octanediol, 1.0% (v/v) glycerol, and 0.5mg/mL nisin. The test conditions of example two above were used, and the results are shown in table 4 below:
table 4 statistical table of test results of anticorrosion system constructed in the third comparative example of the present invention
As can be seen from tables 3 and 4, the preservative systems constructed in example two and control example three both reduced the bacteria concentration by 99.9% and the mold by 90% at day 7, and continued to decrease the number of bacteria within 28 days, both passing the preservative challenge test. However, the aloe fermented raw juice of the third comparative example is turbid after adding related components, and precipitates are generated, so that the activity of removing ABTS & lt + & gt and free radicals is reduced. However, the second embodiment does not have the phenomenon, so that the fermented raw juice can be better preserved by constructing an antiseptic system according to the second embodiment, and the fermented raw juice can be used as an antiseptic matched with the aloe fermented raw juice.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all equivalent changes made by using the contents of the present specification and the drawings, or applied directly or indirectly to the related technical fields, are included in the scope of the present invention.
Claims (10)
1. A preparation method of lactobacillus pentosus aloe fermentation raw stock is characterized by comprising the following steps: the method comprises the following steps:
s1, raw material treatment: squeezing sterilized aloe to obtain juice;
s2, domestication of zymocyte: preparing the aloe juice treated in the step S1 into an aloe seed culture medium, culturing the activated 2-3 generations of lactobacillus pentosus at 30-38 ℃ for 16-24 h, inoculating the lactobacillus pentosus into the aloe seed culture medium according to the volume ratio of 1-3%, and continuously culturing at 30-38 ℃ for 16-24 h to obtain a fermented seed solution for later use;
s3, preparation of fermentation raw pulp: preparing the aloe juice prepared in the step S1 into an aloe fermentation culture medium, inoculating 1-10% by volume of the fermentation seed liquid treated in the step S2 into the aloe fermentation culture medium, and standing and culturing at 30-38 ℃ for 1-20 days;
s4, centrifuging to obtain supernatant, and sterilizing to obtain the fermented raw stock.
2. The method for preparing lactobacillus pentosus aloe fermentation puree as claimed in claim 1, wherein the method comprises the following steps: the aloe is also pretreated by the following steps before sterilization: peeling Aloe, collecting flesh, washing with water, and adding VC for color protection.
3. The method for preparing lactobacillus pentosus aloe fermentation puree as claimed in claim 2, wherein the method comprises the following steps: the mass addition proportion of the VC is 0.01-0.2%.
4. The method for preparing lactobacillus pentosus aloe fermentation puree as claimed in claim 1, wherein the method comprises the following steps: the preparation method of the aloe fermentation medium of the step S3 comprises the following steps: mixing aloe juice with sterilized sucrose-containing solution, and mixing; preferably, the mass ratio of the aloe juice in the aloe fermentation medium is 30-60%; preferably, the mass volume of the sucrose in the sucrose-containing solution accounts for 3-7%.
5. The method for preparing lactobacillus pentosus aloe fermentation puree as claimed in claim 1, wherein the method comprises the following steps: the sterilization in step S4 is performed by filter sterilization.
6. A fermentation broth prepared by the process of any one of claims 1 to 5.
7. The use of the fermented puree of claim 6 for preparing an antioxidant, an anti-allergy agent or a whitening agent.
8. Use of the fermented puree of claim 6 for the preparation of a cosmetic product.
9. The method for preserving the lactobacillus pentosus aloe fermented puree as claimed in claim 6, wherein the method comprises the following steps: the method comprises the following steps: adding polyalcohol into the fermentation raw stock.
10. The saving method according to claim 9, wherein: the storage method specifically comprises the following steps: adding p-hydroxyacetophenone with the mass volume ratio of not more than 1%, hexanediol with the volume ratio of not more than 1% and ethylhexyl glycerol with the volume ratio of not more than 1% into the fermentation raw stock.
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