CN112826076A - Body-building type sugar-cored apple flavor enzyme - Google Patents
Body-building type sugar-cored apple flavor enzyme Download PDFInfo
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- CN112826076A CN112826076A CN202011636793.8A CN202011636793A CN112826076A CN 112826076 A CN112826076 A CN 112826076A CN 202011636793 A CN202011636793 A CN 202011636793A CN 112826076 A CN112826076 A CN 112826076A
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Images
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- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
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- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
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Abstract
The invention relates to a body-building type sugar core apple flavor enzyme which comprises the following preparation steps: the method comprises the following steps of high-low temperature asynchronous whole-cell biological crushing pulping: the method comprises the following steps of: 1) culturing lactobacillus brevis to obtain bacterial suspension; 2) combining multifunctional plasma with ultrasound; high-flux and low-O2And (4) performing high NO circulating secondary fermentation to obtain an apple enzyme stock solution. The invention aims at the technological innovation of powder products in enzyme products, takes sugar-cored Fuji apples as a basic raw material, and utilizes lactobacillus brevis and saccharomyces cerevisiae to perform synergistic fermentation to develop a high-color-value strong-flavor body-building enzyme product.
Description
Technical Field
The invention belongs to the technical field of food, and particularly relates to a body-building type sugar core apple flavor enzyme.
Background
At present, enzyme health care products Linnao appearing in domestic markets are full of eyes, but due to different processing technologies, the enzyme health care products have the problems of low concentration of effective components, low nutrient content, dark color of powdery enzyme, low odor and the like.
The specific problems are as follows:
1. the ferment beverage has low fermentation efficiency and long fermentation time. Harmful bacteria are easy to breed in the fermentation process, and the later-period enzyme beverage is not bright in color.
2. The single strain is fermented, and the finished product has low probiotic richness and active ingredients. The problem of sedimentation is easy to occur due to overlong fermentation production period. Multiple bacteria are subjected to compound fermentation, the optimal fermentation conditions are not completely the same, and certain mutual inhibition exists.
3. The enzyme product has low bioavailability of nutrients of fruits, and the enzyme beverage has insufficient fragrance in the later period.
Through searching, the following patent publications related to the patent application of the invention are found:
1. a mulberry enzyme powder and a preparation method thereof (CN 110037287A), the raw materials are: 70-85% of mulberry enzyme raw pulp and 15.0-30% of resistant dextrin. The mulberry ferment powder is prepared by spraying the mulberry raw juice and is easier to eat by people.
The advantages are that: according to the mulberry enzyme powder, the proper amount of L-arabinose and xylitol are added to promote the absorption of minerals in the mulberry enzyme powder, so that the nutrient elements in the mulberry enzyme powder are absorbed and utilized by a human body to the maximum extent.
The problems are as follows: the single strain is fermented, and the finished product has low probiotic richness and active ingredients. The fermentation mode is preferably anaerobic fermentation, the mulberry enzyme liquid obtained after fermentation for 1-2 years is subjected to residue-liquid separation to obtain the mulberry enzyme raw pulp, and the sedimentation phenomenon is easy to occur due to overlong production period.
And (4) countermeasure: the invention adopts composite fermentation, and improves the fermentation efficiency of the strain through synergistic effect.
2. An edible apple ferment beverage and a preparation method thereof (CN 110403115A), the invention comprises the following raw materials: 40-80 parts of apple enzyme stock solution, 5-15 parts of isomaltooligosaccharide, 1-3 parts of inulin and 0.03-0.05 part of citric acid.
The advantages are that: the edible apple enzyme beverage disclosed by the invention is rich in nutrition, rich in flavor and higher in active substance, and can improve the nutritional value of the product and the additional value of apple resources.
The problems are as follows: firstly, the apples are cleaned, dried by an ozone water machine, peeled, denucleated and sliced, and then are fermented, but although the ozone water machine is used for drying and sterilizing in the pretreatment process, the slicing process is not bacteriostatic, and the later fermentation process is 3-6 months long, so that harmful bacteria are easy to breed. After the apples are cut, the pulp is quickly browned after being exposed in the air, and the invention does not carry out any anti-browning treatment, so that the later-stage ferment beverage is not bright in color.
And (4) countermeasure: the invention adopts high-low temperature asynchronous whole cell biological disruption and bacteriostasis pulping technology, reduces the activity and oxidation degree of enzyme, and greatly protects the original color and nutrient substances of the apples.
3. The invention discloses a fruit enzyme beverage and a preparation method thereof (patent No. CN 110810689A), and the fruit enzyme beverage and the preparation method thereof are prepared by the steps of cleaning, wiping, dicing, mixing, fermenting, filling, refrigerating and finishing.
The advantages are that: the fruit ferment beverage obtained by fully fermenting the pulp, the fermented purified water and the comprehensive rice and bean ferment does not contain artificial pigments, chemical spices and preservatives, has the pure natural characteristic, and improves the taste of the beverage while improving the nutrition by adding the honey.
The problems are as follows: the raw material pretreatment is not carried out with bacteriostasis and color protection treatment. The bioavailability of the original nutrient substances is not high. ③ the smell is not strong enough.
And (4) countermeasure: in the invention, the low-temperature color protection operation is adopted in the raw material treatment and processing processes, so that the activity and the oxidation degree of enzyme are reduced, and the original color and nutrient substances of the apples are protected to a great extent. Secondly, a secondary fermentation composite aroma-enhancing technology is adopted, the content of ester compounds is improved, volatile aglycone is released, and the finished product is rich in ester aroma.
By contrast, the present patent application is substantially different from the above patent publications.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides the body-building type sugar core apple flavor enzyme.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a body-building candied apple flavor enzyme is prepared by the following steps:
the method comprises the following steps of high-low temperature asynchronous whole-cell biological crushing pulping:
1) removing the mildewed part of young or defective sugared apple fruits, cleaning the surface ash layer and soil, keeping the original wild yeast on the epidermis, placing in a compound normal-temperature enzymolysis liquid, wherein the enzymolysis reaction temperature is 23-25 ℃, simultaneously pressurizing to 1.5-1.7 times of atmospheric pressure, reducing the temperature to below 0 ℃ after 5-10min of enzymolysis reaction, and crushing at low temperature to obtain an enzymolysis liquid;
wherein the compound normal-temperature enzymolysis liquid is a mixed aqueous solution of lysozyme, pectinase, cellulase and amylase in a mass ratio of 1:2:1: 1;
2) culturing the bacillus natto to obtain a cultured bacillus natto liquid;
3) in the linkage of high-voltage corona electric field12C6+Treating the bacillus natto by an ion beam biological system: placing the bacillus natto bacterial solution into a pre-sterilized container with glass beads under an aseptic condition, and oscillating to disperse the bacteria; counting by using a blood counting plate; bacterial density is controlled to 10 by diluting bacterial liquid with normal saline7Preparing a bacterial suspension on the seed/seed m L; placing the prepared Bacillus natto strain suspension in a container with a rotor therein, wherein the ion source type is12C6+Ion implantation energy is 10-30keV, and pulse dose is n × 2.6 × 1013ions.cm-2Opening gas valve to make vacuum 7.0-8.2X 10 in the course of injecting arc discharge ion beam-3Pa, adjusting the filament current to be 30-40A, adjusting the magnetic field voltage to be 10-12.5V and adjusting the anode voltage to be 75-100V;
4) diluting the treated bacterial suspension to 108-109After culturing at m L, centrifuging the diluted bacterial suspension at 10000r/min for 10min, and removing bacteria; adding 4 times of anhydrous ethanol into the supernatant for alcohol precipitation, dissolving the supernatant into distilled water with the same volume again, and performing secondary alcohol precipitation to obtain biological fermentation liquor;
5) adding citric acid and calcium chloride to the enzymatic hydrolysate obtained in the step 1), and adding citric acid to the biological fermentation liquid obtained in the step 4: the mass ratio of the enzymolysis liquid is 1:50, and the mass ratio of the citric acid, the calcium chloride and the biological fermentation liquid is 1:1:3, so as to obtain mixed enzymolysis liquid;
6) filtering the skin residue from the mixed enzymolysis liquid to obtain fermentation liquid;
the method comprises the following steps of:
1) culturing lactobacillus brevis to obtain bacterial suspension;
2) multifunctional plasma combined with ultrasound: uniformly coating and airing the bacterial suspension, placing the bacterial suspension in a multifunctional plasma combined ultrasonic system, inputting power of 120W, irradiating at an interval of 3mm, carrying out nitrogen flow of 10L/min, and carrying out treatment for 90 s; the plasma contains active ingredients; adding physiological saline, bacterial suspension: the volume ratio of the physiological saline is 1: 10, preparing the strain into a bacterial suspension; diluting to 1 × 10-5After gradient, uniformly coated on CaCO3Culturing the surface of the culture medium at 30 ℃ for 24 hours; selecting Lactobacillus brevis with largest diameter of transparent circle/colony, and diluting to 10%5Inoculating CFU/ml gradient into the fermentation liquor obtained in the step of step C6);
high-flux and low-O2High NO cycle secondary fermentation:
culturing saccharomyces cerevisiae strains:
firstly, culturing saccharomyces cerevisiae on WL nutrient agar culture medium for 3 days to obtain single colony, and activating the single colony in a test tube added with YPD liquid culture medium for two days;
adding 250 microliters of culture medium into each hole of a 96-hole plate, adding 15 microliters of activated single colonies, adding two holes into each activated single colony, uniformly coating, and culturing for 5 days, and selecting dark brown single colonies as the high-yield glucose-shake enzyme yeast strains; inoculation 106Adding the CFU/ml of saccharomyces cerevisiae into the fermentation liquor treated in the step II 2, and obtaining an apple enzyme stock solution after 5-8 days;
the fermentation process adopts a closed environment which specifically comprises the following steps: 60% NO + 30% N2+ 10% air.
The specific steps of culturing bacillus natto in step # 2) are as follows:
firstly, a preservation number is obtained: bacillus natto of BNCC 185325
② plate culture: preparing plate separation medium, pouring in each plateAdding into culture medium, diluting the bacteria solution to be inoculated, and selecting three diluted bacteria solutions with three concentrations, namely 106CFU/ml、107CFU/ml、108Respectively inoculating CFU/ml into three parallel culture dishes, uniformly coating a bacterial liquid on the surface of a solid culture medium by using a coater in a clockwise or anticlockwise direction, and inversely placing the inoculated culture dishes into a constant-temperature incubator for culturing for 48 hours at 37 ℃;
③ shaking bottles and fermenting culture: picking a single colony on a plate culture medium by using an inoculating needle, inoculating the single colony on a shake flask culture medium, putting a conical flask into a constant-temperature shaking table, and culturing for 24 hours at 37 ℃ and 200 rpm; inoculating the shake flask bacterial liquid cultured for 24 hours into a fermentation medium according to the inoculation proportion of 2%, putting the fermentation medium into a constant-temperature shaking table, and culturing for 48 hours at 37 ℃ and 200rpm to obtain the cultured bacillus natto bacterial liquid.
In the step C3), the container with the rotor is a plate, and 4mL of bacterial suspension is added into each 90mm plate.
The specific culture steps of the lactobacillus brevis in the step II are as follows;
inoculating activated lactobacillus brevis into a fermentation medium in an inoculation amount of 10% (V/V), standing at 30 ℃ for fermentation for 12 hours, supplementing sodium glutamate with a final mass concentration of 100g/L, and continuing fermentation for 48 hours; the fermentation medium comprises the following components in percentage by weight: 0.05% of sodium acetate, 0.05% of magnesium sulfate, 0.05% of manganese sulfate, 0.2% of diammonium citrate, 0.5% of yeast extract, 1% of glucose, 1% of peptone, 1% of beef extract, 1% of agar powder and water as a solvent;
secondly, inoculating the activated seed liquid into a seed culture medium for culturing for 10 hours in an inoculation amount of 10% (V/V) to enable thalli to be in the late stage of logarithmic growth; the seed culture medium comprises the following components in percentage by weight: 0.05% of magnesium sulfate, 0.8% of yeast extract, 0.8% of peptone, 0.1% of tween 80, 2% of glucose and water as a solvent;
③ centrifuging the seed culture solution at 12000r/min for 2min, collecting thallus, washing with sterile normal saline for 3 times, and finally suspending the thallus in the sterile normal saline to make the final concentration of the cells to be 1 × 107CFU/m L to obtain bacterial suspension.
And the steps describedThe lactobacillus brevis in the second step is lactobacillus brevis CICC 20014; or the active ingredient in the second step 2) comprises nitrogen positive ion N + and atomic oxygen O, OH－A free radical.
And, the step of 2) CaCO3The culture medium comprises the following components in percentage by weight: 0.05% of monopotassium phosphate, 0.2% of calcium carbonate, 0.5% of peptone, 1% of glucose, 2% of yeast extract and water as a solvent.
And in the step three, the saccharomyces cerevisiae is saccharomyces cerevisiae CICC 1001.
And the formula of the WL nutrient agar culture medium in the step three is as follows:
the weight percentage is as follows: 0.05% of ferric citrate, 0.2% of sodium chloride, 0.05% of magnesium sulfate heptahydrate, 0.1% of monopotassium phosphate, 2% of agar and water as a solvent.
The invention has the advantages and positive effects that:
1. the invention aims at the technological innovation of powder products in enzyme products, takes sugar-cored Fuji apples as a basic raw material, and utilizes lactobacillus brevis and saccharomyces cerevisiae to perform synergistic fermentation to develop a high-color-value strong-flavor body-building enzyme product.
2. After the apples are cut, the pulp is quickly browned after being exposed in the air, so that the later enzyme beverage is not bright in color. The fermentation period is long, and harmful bacteria are easy to breed. The invention utilizes normal temperature enzymolysis liquid, broad spectrum bacteriostatic lasting biological fermentation liquid with prolonged sterilization effect, and cold crushing treatment to make the cut produce simultaneously with the color-protecting bacteriostatic coating operation. In addition, in the linkage of the high-voltage corona electric field, a 12C6+ ion beam biological system can assist the bacillus natto (BNCC 185325) to produce high-yield homogenous polypeptide (natto gum) and metabolites with bacteriostatic action, so that the bacteriostatic and color-protecting efficiency and effect are improved. Residual oxygen in water is converted into ozone by high-voltage discharge, and atomic oxygen released by the ozone can quickly penetrate cell membranes of cells to kill pathogenic bacteria cells. The contact with oxygen in the air is isolated, the activity and the oxidation degree of enzyme are reduced, and the original color and nutrient substances of the apples are protected to a great extent.
3. The invention solves the problems that the single strain fermentation causes the enrichment degree and the active ingredients of the probiotics in the finished product to be lower and the fermentation production period is overlong and the sedimentation phenomenon is easy to occur. And secondly, the fermentation efficiency of the strains is improved by adopting the composite fermentation through the synergistic effect (firstly, the single-strain fermentation time is generally considered to be longer than that of the composite strains; and secondly, the table 5 shows that the ethyl caprylate, the ethyl acetate and the butyl butyrate ester compounds in the ferment beverage prepared by treating the A (saccharomyces cerevisiae and lactobacillus brevis) are relatively high in content compared with the B (lactobacillus brevis), so that the ferment beverage is rich in ester fragrance, and the composite strain fermentation effect is good. In addition, the acid resistance of lactobacillus brevis is obviously improved, and the enrichment of high-yield GABA and flavonoid aglycone is beneficial to strengthening the antioxidation and anti-aging effects of the ferment.
4. The invention solves the problems of low bioavailability of original nutrient substances in apples and insufficient aroma of enzyme beverage, and adopts high flux and low O2The high NO circulating secondary fermentation composite aroma-enhancing technology enriches sorbitol, improves the content of ester compounds, releases volatile aglycone and ensures that the finished product is rich in ester aroma.
5. The invention adopts high-low temperature asynchronous whole cell biological disruption bacteriostatic pulping technology:
(1) putting the cleaned whole fruit (including pericarp) into normal temperature enzymolysis liquid (lysozyme: pectinase: cellulase: amylase: 1:1) compounded by lysozyme and biological enzyme, wherein the enzymolysis reaction temperature is 25 ℃, and simultaneously pressurizing to 1.5 times of atmospheric pressure, so that the enzymolysis liquid permeates into the deep part of the cell under the action of pressure, the deep layer lasts for a long time, and the cell wall is fully softened and enzymolyzed.
(2) In linkage by high-voltage corona electric field12C6+The ion beam biological system assists bacillus natto (BNCC 185325) to produce high-yield homogenous polypeptide (natto gum) and metabolites with bacteriostatic action, such as 2, 6-dipicolinic acid, subtilisin, gramicidin and the like. The natto gum is easy to swell in water at the later stage and wraps the bacteriostatic product to form hydrogel which can keep a large amount of water in volume but is not dissolved, so that the bactericidal substance is slowly released, and the broad-spectrum bacteriostatic lasting biological fermentation broth with the prolonged bactericidal effect is formed. Additionally using high-voltage dischargeThe residual oxygen in the water is converted into ozone, and the atomic oxygen released by the ozone can quickly penetrate cell membranes of cells, and can be oxidized with protein membranes of pathogenic bacteria and irreversible oxidation with cell enzymes to kill the pathogenic bacteria cells. After the enzymolysis reaction is carried out for 5-10min, the temperature is reduced to below 0 ℃ and the mixture is crushed at low temperature, citric acid, calcium chloride and biological fermentation liquid (1:1:3) are added into the enzymolysis liquid, the whole fruit is subjected to cold crushing treatment in the bacteriostatic enzymolysis liquid, the enzyme is fully combined with the substrate, the enzymolysis reaction is continued, the polysaccharide is converted into monosaccharide, the generation of the cut is carried out simultaneously with the color-protecting bacteriostatic coating operation, the bacteriostatic and color-protecting efficiency and effect are improved, and the contact with oxygen in the air is isolated.
(3) After the enzymolysis reaction is finished, high-oxygen treatment is carried out to inhibit the polymerization of the quinone, then low-temperature low-oxygen complete crushing is carried out to inhibit browning, the purpose of preventing browning is achieved, and finally the skin residue and the like are filtered.
6. The invention adopts multifunctional biological plasma combined with ultrasonic auxiliary primary fermentation:
under the action of multifunctional biological plasma combined with an ultrasonic auxiliary system, lactobacillus brevis (CICC 20014) can carry out primary fermentation on acid compounds such as gamma-aminobutyric acid (GABA) and lactic acid, acetic acid and the like with high yield, and the acid resistance of the lactobacillus brevis is obviously improved by adding 0.5% of sodium glutamate. GABA is a non-protein amino acid with physiological effects of resisting aging, lowering blood pressure, tranquilizing, and promoting sleep, and can activate brain, liver and kidney to achieve body constitution strengthening effect. (multifunctional biological plasma auxiliary system: input power 120W, irradiation interval 3mm, nitrogen flow 10L/min, treatment time 90 s; plasma contains nitrogen positive ion N +, atomic oxygen O, OH－Active components such as free radicals and the like can increase the permeability of microbial cell walls or cell membranes under the assistance of ultrasound, and active ions can excite the self-repairing mechanism of cells after entering the cells to promote the enrichment of gamma-aminobutyric acid. )
In addition, most of the flavonoid compounds in the fruits exist in the form of flavonoid glycoside, most of the flavonoid glycoside can not enter blood through the small intestinal wall in a human body, and the bioavailability is low. The lactobacillus brevis fermentation deglycosylation can convert the flavonoid glycoside into corresponding flavonoid aglycone, so that the flavonoid aglycone is easier to be absorbed by human bodies, the bioavailability of the flavonoid compounds in Fuji apples is greatly improved, and the purpose of oxidation resistance is achieved. The enrichment of high yield GABA and flavonoid aglycone is beneficial to strengthening the antioxidation and anti-aging effects of the ferment.
7. The invention adopts a secondary fermentation composite aroma-enhancing technology:
using high flux low O2High NO circulating fermentation high-yield beta-glucosidase saccharomyces cerevisiae (CICC 1001) is subjected to secondary fermentation (both bifidobacterium breve and saccharomyces cerevisiae are facultative anaerobes). The fermentation process adopts a closed environment which specifically comprises the following steps: 60% NO + 30% N2+ 10% air. The anaerobic glycolysis of pulp and ethanol synthesis can be activated in a low-oxygen environment, so that the yield of ester compounds such as ethyl caprylate, ethyl acetate, butyl butyrate and the like is increased, and the enzyme beverage is rich in ester fragrance.
As shown in figure 1, the sugar content, particularly the sorbitol content, of the sugar-cored apples is much higher than that of common apples, the spatial structure of sorbitol dehydrogenase is changed under the action of NO, cysteine Cys in a catalytic domain related to the catalytic function is easy to have nitrosation reaction with NO, so that most of the catalytic function of the sorbitol dehydrogenase is inhibited, a large amount of sorbitol is accumulated, and the sorbitol is used as a nutritive sweetener, is not controlled by insulin, has a health-care function and is suitable for being eaten by diabetics.
Most of flavor compounds in the Fuji apples exist in a tasteless non-volatile glycoside form, so that the flavor of the enzyme product is not strong enough. The beta-glucosidase with high yield by saccharomyces cerevisiae fermentation can hydrolyze the glucoside fragrance precursors to release volatile aglycone, so that the enzyme beverage is rich in sweet and mellow fragrance.
Drawings
FIG. 1 is a diagram showing the metabolism of sorbitol in fruits under the action of nitric oxide.
Detailed Description
The present invention is further illustrated by the following examples, which are intended to be illustrative, not limiting and are not intended to limit the scope of the invention.
The raw materials used in the invention are all conventional commercial products if no special description is provided, the method used in the invention is all conventional methods in the field if no special description is provided, and the mass of all the materials used in the invention is the conventional use mass.
A body-building candied apple flavor enzyme is prepared by the following steps:
the method comprises the following steps of high-low temperature asynchronous whole-cell biological crushing pulping:
1) removing the mildewed part of young or defective sugared apple fruits, cleaning the surface ash layer and soil, keeping the original wild yeast on the epidermis, placing in a compound normal-temperature enzymolysis liquid, wherein the enzymolysis reaction temperature is 23-25 ℃, simultaneously pressurizing to 1.5-1.7 times of atmospheric pressure, reducing the temperature to below 0 ℃ after 5-10min of enzymolysis reaction, and crushing at low temperature to obtain an enzymolysis liquid;
wherein the compound normal-temperature enzymolysis liquid is a mixed aqueous solution of lysozyme, pectinase, cellulase and amylase in a mass ratio of 1:2:1: 1;
2) culturing the bacillus natto to obtain a cultured bacillus natto liquid;
3) in the linkage of high-voltage corona electric field12C6+Treating the bacillus natto by an ion beam biological system: placing the bacillus natto bacterial solution into a pre-sterilized container with glass beads under an aseptic condition, and oscillating to disperse the bacteria; counting by using a blood counting plate; bacterial density is controlled to 10 by diluting bacterial liquid with normal saline7Preparing a bacterial suspension on the seed/seed m L; placing the prepared Bacillus natto strain suspension in a container with a rotor therein, wherein the ion source type is12C6+Ion implantation energy is 10-30keV, and pulse dose is n × 2.6 × 1013ions.cm-2Opening gas valve to make vacuum 7.0-8.2X 10 in the course of injecting arc discharge ion beam-3Pa, adjusting the filament current to be 30-40A, adjusting the magnetic field voltage to be 10-12.5V and adjusting the anode voltage to be 75-100V;
4) diluting the treated bacterial suspension to 108-109After culturing at m L, centrifuging the diluted bacterial suspension at 10000r/min for 10min, and removing bacteria; adding 4 times volume of anhydrous ethanol into the supernatant to precipitate ethanol, and dissolving againDissolving the fermented liquid in distilled water with the same volume, and performing secondary alcohol precipitation to obtain biological fermentation liquid;
5) adding citric acid and calcium chloride to the enzymatic hydrolysate obtained in the step 1), and adding citric acid to the biological fermentation liquid obtained in the step 4: the mass ratio of the enzymolysis liquid is 1:50, and the mass ratio of the citric acid, the calcium chloride and the biological fermentation liquid is 1:1:3, so as to obtain mixed enzymolysis liquid;
6) filtering the skin residue from the mixed enzymolysis liquid to obtain fermentation liquid;
the method comprises the following steps of:
1) culturing lactobacillus brevis to obtain bacterial suspension;
2) multifunctional plasma combined with ultrasound: uniformly coating and airing the bacterial suspension, placing the bacterial suspension in a multifunctional plasma combined ultrasonic system, inputting power of 120W, irradiating at an interval of 3mm, carrying out nitrogen flow of 10L/min, and carrying out treatment for 90 s; the plasma contains active ingredients; adding physiological saline, bacterial suspension: the volume ratio of the physiological saline is 1: 10, preparing the strain into a bacterial suspension; diluting to 1 × 10-5After gradient, uniformly coated on CaCO3Culturing the surface of the culture medium at 30 ℃ for 24 hours; selecting Lactobacillus brevis with largest diameter of transparent circle/colony, and diluting to 10%5Inoculating CFU/ml gradient into the fermentation liquor obtained in the step of step C6);
high-flux and low-O2High NO cycle secondary fermentation:
culturing saccharomyces cerevisiae strains:
firstly, culturing saccharomyces cerevisiae on WL nutrient agar culture medium for 3 days to obtain single colony, and activating the single colony in a test tube added with YPD liquid culture medium for two days;
adding 250 microliters of culture medium into each hole of a 96-hole plate, adding 15 microliters of activated single colonies, adding two holes into each activated single colony, uniformly coating, and culturing for 5 days, and selecting dark brown single colonies as the high-yield glucose-shake enzyme yeast strains; inoculation 106Adding the CFU/ml of saccharomyces cerevisiae into the fermentation liquor treated in the step II 2, and obtaining an apple enzyme stock solution after 5-8 days;
the fermentation process adopts a closed environment which specifically comprises the following steps: 60% NO + 30% N2+ 10% air.
Preferably, the step of preparing the bacillus natto in step # 2) comprises the following steps:
firstly, a preservation number is obtained: bacillus natto of BNCC 185325
② plate culture: preparing a plate separation culture medium, pouring the culture medium into each plate, diluting the bacteria liquid to be inoculated, and selecting three diluted bacteria liquids with three concentrations, namely 106CFU/ml、107CFU/ml、108Respectively inoculating CFU/ml into three parallel culture dishes, uniformly coating a bacterial liquid on the surface of a solid culture medium by using a coater in a clockwise or anticlockwise direction, and inversely placing the inoculated culture dishes into a constant-temperature incubator for culturing for 48 hours at 37 ℃;
③ shaking bottles and fermenting culture: picking a single colony on a plate culture medium by using an inoculating needle, inoculating the single colony on a shake flask culture medium, putting a conical flask into a constant-temperature shaking table, and culturing for 24 hours at 37 ℃ and 200 rpm; inoculating the shake flask bacterial liquid cultured for 24 hours into a fermentation medium according to the inoculation proportion of 2%, putting the fermentation medium into a constant-temperature shaking table, and culturing for 48 hours at 37 ℃ and 200rpm to obtain the cultured bacillus natto bacterial liquid.
Preferably, the container with the rotor in step C3) is a plate, and 4mL of the bacterial suspension is added into each 90mm plate.
Preferably, the method comprises the following specific culture steps of the lactobacillus brevis;
inoculating activated lactobacillus brevis into a fermentation medium in an inoculation amount of 10% (V/V), standing at 30 ℃ for fermentation for 12 hours, supplementing sodium glutamate with a final mass concentration of 100g/L, and continuing fermentation for 48 hours; the fermentation medium comprises the following components in percentage by weight: 0.05% of sodium acetate, 0.05% of magnesium sulfate, 0.05% of manganese sulfate, 0.2% of diammonium citrate, 0.5% of yeast extract, 1% of glucose, 1% of peptone, 1% of beef extract, 1% of agar powder and water as a solvent;
secondly, inoculating the activated seed liquid into a seed culture medium for culturing for 10 hours in an inoculation amount of 10% (V/V) to enable thalli to be in the late stage of logarithmic growth; the seed culture medium comprises the following components in percentage by weight: 0.05% of magnesium sulfate, 0.8% of yeast extract, 0.8% of peptone, 0.1% of tween 80, 2% of glucose and water as a solvent;
③ centrifuging the seed culture solution at 12000r/min for 2min, collecting thallus, washing with sterile normal saline for 3 times, and finally suspending the thallus in the sterile normal saline to make the final concentration of the cells to be 1 × 107CFU/m L to obtain bacterial suspension.
Preferably, the lactobacillus brevis in the step 1) is lactobacillus brevis CICC 20014; or the active ingredient in the second step 2) comprises nitrogen positive ion N + and atomic oxygen O, OH－A free radical.
Preferably, the step is implemented by using CaCO in the step 2)3The culture medium comprises the following components in percentage by weight: 0.05% of monopotassium phosphate, 0.2% of calcium carbonate, 0.5% of peptone, 1% of glucose, 2% of yeast extract and water as a solvent.
Preferably, the saccharomyces cerevisiae in the step three is saccharomyces cerevisiae CICC 1001.
Preferably, the formula of the WL nutrient agar culture medium in the step three is as follows:
the weight percentage is as follows: 0.05% of ferric citrate, 0.2% of sodium chloride, 0.05% of magnesium sulfate heptahydrate, 0.1% of monopotassium phosphate, 2% of agar and water as a solvent.
Specifically, the preparation and detection are as follows:
the preparation process of the body-building candied apple flavor enzyme provided by the invention can be as follows:
raw material screening and cleaning → pressurized soaking → high and low temperature asynchronous whole cell biological disruption and bacteriostasis pulping → filtration → cooling inoculation → multifunctional biological plasma combines ultrasonic auxiliary primary fermentation → secondary fermentation composite flavor enhancement technology → sugar adjustment → index detection → low temperature storage.
More specifically, the preparation method of the body-building candied apple flavor enzyme comprises the following steps:
1. high-low temperature asynchronous whole cell biological disruption pulping:
(1) removing mildew from inferior fruits, cleaning to remove ash layer and soil on the surface, placing original wild yeast on epidermis in a compound normal temperature enzymolysis liquid (lysozyme: pectinase: cellulase: amylase: 1:1), wherein the enzymolysis reaction temperature is 25 ℃, and simultaneously pressurizing to 1.5 times of atmospheric pressure.
(2) Culturing Bacillus natto
The preservation number is: BNCC 185325
② plate culture: preparing a certain amount of plate separation culture medium, pouring 20ml of culture medium into each plate, diluting the to-be-inoculated solution into proper concentrations according to a certain proportion, selecting three kinds of diluted bacterial solutions with different concentrations to be respectively inoculated into three parallel culture plates, wherein the inoculation amount of each plate is 20 microliters of bacterial solution, uniformly coating the bacterial solutions on the surface of the solid culture medium in a clockwise or anticlockwise direction by using a coater, and inversely placing the inoculated culture plates into a constant-temperature incubator to be cultured for 48 hours at 37 ℃.
③ shaking bottles and fermenting culture: and (3) picking a single colony on the plate culture medium by using an inoculating needle to inoculate the single colony to a shake flask culture medium, putting the conical flask into a constant-temperature shaking table, and culturing for 24 hours at 37 ℃ and 200 rpm. Inoculating the shake flask bacterial liquid cultured for 24h into a fermentation medium according to the proportion of 2%, putting the conical flask containing the fermentation liquid into a constant temperature shaking table, and culturing for 48h at 37 ℃ and 200 rpm.
(3) In the linkage of high-voltage corona electric field12C6+Treating the bacillus natto by an ion beam biological system: the growth phase bacterial liquid is put into a pre-sterilized triangular flask with glass beads under aseptic condition, and the vibration is carried out to disperse the thalli. Counting was performed with a hemocytometer. Bacterial density is controlled to 10 by diluting bacterial liquid with normal saline7The bacterial suspension is prepared on m L. Taking 4m L prepared Bacillus natto strain suspension in a 90mm plate with a rotor, wherein the ion source type is12C6+Ion implantation energy is 10-30keV, and pulse dose is n × 2.6 × 1013ions.cm-2The vacuum of the gas valve is 7.0-8.2X 10 during the injection process of the arc discharge ion beam-3Pa, filament current is adjusted to be 30-40A, magnetic field voltage is adjusted to be 10-12.5V, and anode voltage is 75-100V.
(4) And (3) diluting the bacterial suspension to a proper mass concentration, centrifuging the fermentation liquor at 10000r/min for 10min, and removing the bacteria. And adding 4 times of volume of absolute ethyl alcohol into the supernatant for alcohol precipitation, then dissolving the supernatant into distilled water with the same volume again, and then carrying out secondary alcohol precipitation to obtain the biological fermentation liquor.
(5) After the enzymolysis reaction is carried out for 5-10min, the temperature is reduced to below 0 ℃ and the mixture is crushed at low temperature, citric acid, calcium chloride and biological fermentation liquid (1:1:3) are added into the enzymolysis liquid, and the whole fruit is subjected to cold crushing treatment in the bacteriostatic enzymolysis liquid.
(6) And filtering the skin dregs.
2. Primary fermentation:
(1) culturing a strain of lactobacillus brevis (CICC 20014):
inoculating activated lactobacillus brevis into a 500m L triangular flask containing 50m L fermentation medium in an inoculation amount of 10% (V/V), standing at 30 ℃ for fermentation for 12h, supplementing 100g/L sodium glutamate, and continuing fermentation for 48 h.
② the activated seed liquid is inoculated into a seed culture medium for 10 hours with the inoculation amount of 10 percent (V/V) to culture, so that the thalli are in the late stage of logarithmic growth.
③ taking 1m L the seed culture solution, centrifuging at 12000r/min for 2min, collecting thallus, washing with sterile normal saline for 3 times, and finally suspending the thallus in the sterile normal saline to make the final concentration of the cells 1X 107CFU/m L。
(2) The multifunctional plasma combined ultrasonic system comprises: uniformly coating and airing 10 mu L of the bacterial suspension, placing the bacterial suspension in a multifunctional plasma combined ultrasonic system, inputting 120W of power, irradiating at an interval of 3mm, carrying out nitrogen flow at a rate of 10L/min, and carrying out treatment for 90 s; the plasma contains N + of nitrogen positive ion and O, OH of atomic oxygen－Free radicals and the like. Adding 100 μ L physiological saline, and making into bacterial suspension. Diluting to 1 × 10-5After the gradient, 100. mu.L of the suspension was uniformly applied onto the surface of CaCO3 medium and cultured at 30 ℃ for 24 hours. Selecting Lactobacillus brevis 10 with larger diameter of transparent circle/bacterial colony5CFU/ml was inoculated into the fermentation broth.
3. High flux low O2High NO cycle secondary fermentation:
(1) culturing saccharomyces cerevisiae (CICC 1001) strain:
firstly, saccharomyces cerevisiae is cultured on WL nutrient agar culture medium (0.05 percent of ferric citrate, 0.2 percent of sodium chloride, 0.05 percent of magnesium sulfate heptahydrate, 0.1 percent of potassium dihydrogen phosphate and 2 percent of agar water in percentage by weight) for 3 days to obtain single colony, and the single colony is activated in a test tube added with YPD liquid culture medium for two days.
② 250 microliter of culture medium is added into each hole of a 96-hole plate, 15 microliter of activated single colony is added, and two holes are added into each activated single colony for uniform coating. After 5 days of culture, a dark brown-black single colony is selected as the high-yield glucose-shake enzyme yeast strain. Inoculation 106Adding the CFU/ml of saccharomyces cerevisiae into the fermentation liquor, and obtaining the apple ferment stock solution after 5-8 days.
(2) The fermentation process adopts a closed environment which specifically comprises the following steps: 60% NO + 30% N2+ 10% air.
The correlation detection results of the invention are as follows:
1. natto gum yield meter
The two groups of selected strains are subcultured and fermented, and the yield is measured. The fermentation results are shown in Table 1, and can be obtained by high-voltage corona electric field linkage12C6+The bacillus natto with the auxiliary effect of the ion beam biological system can be subcultured, and the excellent character of high yield of the natto gum can be well maintained. The table below shows the yield of natto gum obtained by culturing 25ml fermentation liquor at 37 ℃ and 200rpm for 48 h.
TABLE 1 natto gum yield Table
2. The color of the pulp (A) prepared by the high-low temperature asynchronous whole-cell biological crushing pulping technology is compared with that of the pulp (B) prepared without the high-low temperature asynchronous whole-cell biological crushing pulping technology, and the result is shown in Table 2, which shows that the high-low temperature asynchronous whole-cell biological crushing pulping technology has the color protection effect on the apple ferment. (L)*The larger the value, the higher the brightness)
TABLE 2 influence of different treatments on the Brightness of candied apple pulp
3. Gamma-aminobutyric acid (GABA) output meter
The two groups of selected strains are subcultured and fermented, and the yield is measured. The fermentation result is shown in table 3, the lactobacillus brevis subjected to the action of the multifunctional biological plasma combined with the ultrasonic auxiliary system can be subcultured, and the excellent character of high GABA yield can be well reserved. The following table shows the GABA yields obtained by culturing 20ml of fermentation broth at 37 ℃ and 200rpm for 36 h.
TABLE 3 Gamma-aminobutyric acid (GABA) yield Table
4. Sorbitol content scale
Ferment (A) prepared by secondary fermentation of saccharomyces cerevisiae by high-flux low-O2 high-NO circulating fermentation and ferment prepared by non-high-flux low-O2The comparison of the sorbitol content of the ferment (B) prepared by the secondary fermentation of the high NO circulating fermentation Saccharomyces cerevisiae shows that, as shown in Table 4, a large amount of sorbitol is accumulated in the ferment by the secondary fermentation of the high-flux low-O2 high NO circulating fermentation Saccharomyces cerevisiae, and the sorbitol is used as a nutritive sweetener, is not controlled by insulin, has a health care function, and is suitable for being eaten by diabetic patients.
TABLE 4 sorbitol content table
5. GC-MS gas chromatography mass spectrometry combined instrument analysis:
using high flux low O2Ferment (A) prepared by secondary fermentation of high-NO circulating fermentation saccharomyces cerevisiae and ferment prepared by non-adopted high-flux low-O2The spectra of the enzyme (B) prepared by the secondary fermentation of high NO circulating fermentation Saccharomyces cerevisiae were compared, as shown in Table 5, using high NOLow flux of O2The high NO circulating fermentation saccharomyces cerevisiae is subjected to secondary fermentation to increase the relative content of ethyl caprylate, ethyl acetate and butyl butyrate ester compounds, so that the enzyme beverage is rich in ester fragrance.
TABLE 5 fragrance ingredient Table
Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the embodiments disclosed.
Claims (8)
1. The body-building candied apple flavor enzyme is characterized in that: the preparation steps are as follows:
the method comprises the following steps of high-low temperature asynchronous whole-cell biological crushing pulping:
1) removing the mildewed part of young or defective sugared apple fruits, cleaning the surface ash layer and soil, keeping the original wild yeast on the epidermis, placing in a compound normal-temperature enzymolysis liquid, wherein the enzymolysis reaction temperature is 23-25 ℃, simultaneously pressurizing to 1.5-1.7 times of atmospheric pressure, reducing the temperature to below 0 ℃ after 5-10min of enzymolysis reaction, and crushing at low temperature to obtain an enzymolysis liquid;
wherein the compound normal-temperature enzymolysis liquid is a mixed aqueous solution of lysozyme, pectinase, cellulase and amylase in a mass ratio of 1:2:1: 1;
2) culturing the bacillus natto to obtain a cultured bacillus natto liquid;
3) in the linkage of high-voltage corona electric field12C6+Treating the bacillus natto by an ion beam biological system: placing the bacillus natto bacterial solution into a pre-sterilized container with glass beads under an aseptic condition, and oscillating to disperse the bacteria; counting by using a blood counting plate; bacterial density is controlled to 10 by diluting bacterial liquid with normal saline7Preparing a bacterial suspension on the seed/seed m L; taking prepared Bacillus nattoSuspending the liquid in a container with a rotor therein, the ion source being of the type12C6+Ion implantation energy is 10-30keV, and pulse dose is n × 2.6 × 1013ions.cm-2Opening gas valve to make vacuum 7.0-8.2X 10 in the course of injecting arc discharge ion beam- 3Pa, adjusting the filament current to be 30-40A, adjusting the magnetic field voltage to be 10-12.5V and adjusting the anode voltage to be 75-100V;
4) diluting the treated bacterial suspension to 108-109After culturing at m L, centrifuging the diluted bacterial suspension at 10000r/min for 10min, and removing bacteria; adding 4 times of anhydrous ethanol into the supernatant for alcohol precipitation, dissolving the supernatant into distilled water with the same volume again, and performing secondary alcohol precipitation to obtain biological fermentation liquor;
5) adding citric acid and calcium chloride to the enzymatic hydrolysate obtained in the step 1), and adding citric acid to the biological fermentation liquid obtained in the step 4: the mass ratio of the enzymolysis liquid is 1:50, and the mass ratio of the citric acid, the calcium chloride and the biological fermentation liquid is 1:1:3, so as to obtain mixed enzymolysis liquid;
6) filtering the skin residue from the mixed enzymolysis liquid to obtain fermentation liquid;
the method comprises the following steps of:
1) culturing lactobacillus brevis to obtain bacterial suspension;
2) multifunctional plasma combined with ultrasound: uniformly coating and airing the bacterial suspension, placing the bacterial suspension in a multifunctional plasma combined ultrasonic system, inputting power of 120W, irradiating at an interval of 3mm, carrying out nitrogen flow of 10L/min, and carrying out treatment for 90 s; the plasma contains active ingredients; adding physiological saline, bacterial suspension: the volume ratio of the physiological saline is 1: 10, preparing the strain into a bacterial suspension; diluting to 1 × 10-5After gradient, uniformly coated on CaCO3Culturing the surface of the culture medium at 30 ℃ for 24 hours; selecting Lactobacillus brevis with largest diameter of transparent circle/colony, and diluting to 10%5Inoculating CFU/ml gradient into the fermentation liquor obtained in the step of step C6);
high-flux and low-O2High NO cycle secondary fermentation:
culturing saccharomyces cerevisiae strains:
firstly, culturing saccharomyces cerevisiae on WL nutrient agar culture medium for 3 days to obtain single colony, and activating the single colony in a test tube added with YPD liquid culture medium for two days;
adding 250 microliters of culture medium into each hole of a 96-hole plate, adding 15 microliters of activated single colonies, adding two holes into each activated single colony, uniformly coating, and culturing for 5 days, and selecting dark brown single colonies as the high-yield glucose-shake enzyme yeast strains; inoculation 106Adding the CFU/ml of saccharomyces cerevisiae into the fermentation liquor treated in the step II 2, and obtaining an apple enzyme stock solution after 5-8 days;
the fermentation process adopts a closed environment which specifically comprises the following steps: 60% NO + 30% N2+ 10% air.
2. The healthy and sweet apple-flavored ferment according to claim 1, wherein: the method comprises the following specific steps of culturing the bacillus natto in the step I2):
firstly, a preservation number is obtained: bacillus natto of BNCC 185325
② plate culture: preparing a plate separation culture medium, pouring the culture medium into each plate, diluting the bacteria liquid to be inoculated, and selecting three diluted bacteria liquids with three concentrations, namely 106CFU/ml、107CFU/ml、108Respectively inoculating CFU/ml into three parallel culture dishes, uniformly coating a bacterial liquid on the surface of a solid culture medium by using a coater in a clockwise or anticlockwise direction, and inversely placing the inoculated culture dishes into a constant-temperature incubator for culturing for 48 hours at 37 ℃;
③ shaking bottles and fermenting culture: picking a single colony on a plate culture medium by using an inoculating needle, inoculating the single colony on a shake flask culture medium, putting a conical flask into a constant-temperature shaking table, and culturing for 24 hours at 37 ℃ and 200 rpm; inoculating the shake flask bacterial liquid cultured for 24 hours into a fermentation medium according to the inoculation proportion of 2%, putting the fermentation medium into a constant-temperature shaking table, and culturing for 48 hours at 37 ℃ and 200rpm to obtain the cultured bacillus natto bacterial liquid.
3. The healthy and sweet apple-flavored ferment according to claim 1, wherein: the method comprises the step of mixing the bacterial suspension solution and the liquid suspension solution in the step 3), wherein a container with a rotor is a plate, and 4mL of bacterial suspension solution is added into each 90mm plate.
4. The healthy and sweet apple-flavored ferment according to claim 1, wherein: the method comprises the following specific culture steps of the lactobacillus brevis;
inoculating activated lactobacillus brevis into a fermentation medium in an inoculation amount of 10% (V/V), standing at 30 ℃ for fermentation for 12 hours, supplementing sodium glutamate with a final mass concentration of 100g/L, and continuing fermentation for 48 hours; the fermentation medium comprises the following components in percentage by weight: 0.05% of sodium acetate, 0.05% of magnesium sulfate, 0.05% of manganese sulfate, 0.2% of diammonium citrate, 0.5% of yeast extract, 1% of glucose, 1% of peptone, 1% of beef extract, 1% of agar powder and water as a solvent;
secondly, inoculating the activated seed liquid into a seed culture medium for culturing for 10 hours in an inoculation amount of 10% (V/V) to enable thalli to be in the late stage of logarithmic growth; the seed culture medium comprises the following components in percentage by weight: 0.05% of magnesium sulfate, 0.8% of yeast extract, 0.8% of peptone, 0.1% of tween 80, 2% of glucose and water as a solvent;
③ centrifuging the seed culture solution at 12000r/min for 2min, collecting thallus, washing with sterile normal saline for 3 times, and finally suspending the thallus in the sterile normal saline to make the final concentration of the cells to be 1 × 107CFU/m L to obtain bacterial suspension.
5. The healthy and sweet apple-flavored ferment according to claim 1, wherein: the method comprises the following steps of 1), wherein the lactobacillus brevis is lactobacillus brevis CICC 20014; or the active ingredient in the second step 2) comprises nitrogen positive ion N + and atomic oxygen O, OH－A free radical.
6. The healthy and sweet apple-flavored ferment according to claim 1, wherein: the step of 2) preparing CaCO3The culture medium comprises the following components in percentage by weight: 0.05% of monopotassium phosphate, 0.2% of calcium carbonate, 0.5% of peptone, 1% of glucose, 2% of yeast extract and water as a solvent.
7. The healthy and sweet apple-flavored ferment according to claim 1, wherein: and step three, the saccharomyces cerevisiae is saccharomyces cerevisiae CICC 1001.
8. The healthy and centered apple-flavored ferment according to any one of claims 1 to 7, wherein: the formula of the WL nutrient agar culture medium in the step three is as follows:
the weight percentage is as follows: 0.05% of ferric citrate, 0.2% of sodium chloride, 0.05% of magnesium sulfate heptahydrate, 0.1% of monopotassium phosphate, 2% of agar and water as a solvent.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116555074A (en) * | 2023-03-13 | 2023-08-08 | 广东悦创生物科技有限公司 | Lactobacillus brevis JT1 and application thereof in preparation of hypoglycemic food and drug |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000045653A1 (en) * | 1999-01-22 | 2000-08-10 | Arne Rytz | Fermented beverage and method for its production |
| CN104152361A (en) * | 2014-03-04 | 2014-11-19 | 西北农林科技大学 | Screening method of yeast strains for highly producing beta-glucosaccharase |
| WO2016028483A1 (en) * | 2014-08-21 | 2016-02-25 | Hsu Shantung | Active fermentation and fermented drinks and products |
| CN106721797A (en) * | 2016-12-06 | 2017-05-31 | 上海应用技术大学 | A kind of apple ferment and preparation method and application |
| CN109793133A (en) * | 2018-12-10 | 2019-05-24 | 国投中鲁果汁股份有限公司 | Spleen benefiting and stimulating the appetite hawthorn snow pear juice and preparation method |
| CN110964760A (en) * | 2018-09-28 | 2020-04-07 | 天津科技大学 | Mutagenesis breeding method of high-yield gamma-aminobutyric acid strain |
| CN111109591A (en) * | 2019-10-25 | 2020-05-08 | 新疆红旗坡农业发展集团有限公司 | Intestinal microecology high-efficiency reconstruction type apple enzyme and processing technology |
-
2020
- 2020-12-31 CN CN202011636793.8A patent/CN112826076A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000045653A1 (en) * | 1999-01-22 | 2000-08-10 | Arne Rytz | Fermented beverage and method for its production |
| CN104152361A (en) * | 2014-03-04 | 2014-11-19 | 西北农林科技大学 | Screening method of yeast strains for highly producing beta-glucosaccharase |
| WO2016028483A1 (en) * | 2014-08-21 | 2016-02-25 | Hsu Shantung | Active fermentation and fermented drinks and products |
| CN106721797A (en) * | 2016-12-06 | 2017-05-31 | 上海应用技术大学 | A kind of apple ferment and preparation method and application |
| CN110964760A (en) * | 2018-09-28 | 2020-04-07 | 天津科技大学 | Mutagenesis breeding method of high-yield gamma-aminobutyric acid strain |
| CN109793133A (en) * | 2018-12-10 | 2019-05-24 | 国投中鲁果汁股份有限公司 | Spleen benefiting and stimulating the appetite hawthorn snow pear juice and preparation method |
| CN111109591A (en) * | 2019-10-25 | 2020-05-08 | 新疆红旗坡农业发展集团有限公司 | Intestinal microecology high-efficiency reconstruction type apple enzyme and processing technology |
Non-Patent Citations (1)
| Title |
|---|
| 新吉乐图: "电晕电场-离子束符合诱变对纳豆芽孢杆菌产γ-聚谷氨酸产量的研究", 《中国优秀博硕士学位论文全文数据库（硕士）基础科学辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116555074A (en) * | 2023-03-13 | 2023-08-08 | 广东悦创生物科技有限公司 | Lactobacillus brevis JT1 and application thereof in preparation of hypoglycemic food and drug |
| CN116555074B (en) * | 2023-03-13 | 2023-10-27 | 广东悦创生物科技有限公司 | Lactobacillus brevis JT1 and application thereof in preparation of hypoglycemic drugs |
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