CN107788517B - Method for enhancing enzyme efficacy by using lactobacillus bulgaricus - Google Patents
Method for enhancing enzyme efficacy by using lactobacillus bulgaricus Download PDFInfo
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Images
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
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- Biotechnology (AREA)
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- Tropical Medicine & Parasitology (AREA)
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- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
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- Preparation Of Fruits And Vegetables (AREA)
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Abstract
The invention discloses a method for enhancing the efficacy of ferment by using lactobacillus bulgaricus, and belongs to the technical field of fermented foods. According to the invention, saccharomyces cerevisiae N85, gluconacetobacter gluconicum Q1 and lactobacillus delbrueckii subsp bulgaricus B1 are added into the fruit and vegetable raw materials, so that the probiotic effect of the ferment is enhanced, the reducing power of the prepared ferment is improved by 43.3% compared with that of a control group, the DPPH clearance is also improved from 88.25% to 93.09%, the lactic acid content is improved by 36.1%, and the acetic acid content is improved by 35.2%. Compared with a control group, the protease activity is improved by 11.9%, the content of volatile substances is increased, the content of ethyl acetate is improved by 85.1%, the content of amyl acetate is improved by 89.1%, so that the ferment has special fruit fragrance, and the bacteriostatic ability of the ferment is improved.
Description
Technical Field
The invention relates to a method for enhancing the efficacy of ferment by using lactobacillus bulgaricus, and belongs to the technical field of fermented foods.
Background
The plant enzyme is a microbial preparation which is prepared by fermenting plants such as vegetables and fruits serving as raw materials together with multiple microorganisms such as saccharomycetes, acetic acid bacteria and lactobacillus and contains abundant enzymes, vitamins, mineral substances and multiple secondary metabolites. Plant enzymes are also known as ferments. The ferment is different from a common enzyme preparation, and is a health care product with high market value due to the functions of promoting digestion, relaxing bowel, resisting oxidation, inhibiting bacteria, inhibiting tumor and the like. Many existing natural plant enzymes have many limiting factors, such as easy infectious microbes, poor taste or flavor, long fermentation period, difficulty in controlling product quality, and great influence of environment. Therefore, the enzyme preparation method for improving the oxidation resistance of the product, improving the taste and flavor of the enzyme and enhancing the bacteriostatic action is more beneficial to the industrial production of the enzyme.
Disclosure of Invention
The first purpose of the invention is to provide a method for simultaneously improving functional factors and flavor of fruit and vegetable ferment, which is to inoculate saccharomycetes, acetobacter gluconicum and lactic acid bacteria into ferment taking fruits and vegetables as raw materials for fermentation.
In one embodiment of the invention, the Saccharomyces cerevisiae is Saccharomyces cerevisiae N85, Acetobacter gluconicum Q1 and Lactobacillus bulgaricus subsp. bulgaricus strain B1, B2.
The second purpose is to provide a probiotic enzyme which is prepared by taking vegetables and fruits as raw materials and inoculating saccharomycetes, acetobacter gluconicum and lactic acid bacteria for co-fermentation.
In one embodiment of the invention, the probiotic ferment is prepared by taking vegetables and fruits as raw materials and inoculating saccharomyces cerevisiae, acetobacter gluconicum and lactobacillus delbrueckii for co-fermentation.
In one embodiment of the invention, the vegetables and fruits include dragon fruit, apple, tomato, and soybean,
in one embodiment of the invention, the Saccharomyces cerevisiae is Saccharomyces cerevisiae N85, the Acetobacter xylinum is Acetobacter xylinum Q1, and the Lactobacillus is Lactobacillus delbrueckii subsp. bulgaricus B1 or Lactobacillus delbrueckii subsp. bulgaricus B2.
In one embodiment of the invention, Saccharomyces cerevisiae N85 was an industrial strain and was disclosed in 2016, entitled "research on Low-yield Ethyl carbamate by Yeast for brewing yellow wine for Metabolic engineering".
In one embodiment of the invention, the acetobacter gluconicum Q1 is deposited in the chinese type culture collection with the deposit number of CCTCC M2014353 at 23.7.2014.
In one embodiment of the invention, the lactobacillus delbrueckii subsp bulgaricus B1 has been deposited in the chinese type culture collection at 26.6.2017 with the deposition number CCTCC M2017370 and the deposition address wuhan, wuhan university.
In one embodiment of the invention, the Lactobacillus delbrueckii subspecies bulgaricus B2 is derived from the resource library of Lactobacillus species in the Key laboratory of the department of Biotechnology and engineering education, inner Mongolia university of agricultural university, and has the accession number LABCIMAU 20094 DBTE ML 12-2-2.
In one embodiment of the present invention, the ferment is prepared by the following steps: (1) taking dragon fruits, apples, tomatoes and soybeans as raw materials, adding 15-20% of glucose by mass of the raw materials, adding 100% of water by mass of the raw materials, and fully and uniformly stirring; (2) inoculating 1% of yeast and 2% of acetic acid bacteria by volume, culturing at a constant temperature of 35-37 ℃, stirring once every 20-24 hours, fermenting to 3-4 days, inoculating 3-5% of lactic acid bacteria, culturing at a constant temperature of 28-30 ℃ in a sealed manner, and fermenting to 12-15 days to finish fermentation.
The third purpose of the invention is to provide a preparation method of ferment efficacy, which comprises the following steps: (1) taking dragon fruits, apples, tomatoes and soybeans as raw materials, adding 15-20% of glucose by mass of the raw materials, adding 100% of water by mass of the raw materials, and fully and uniformly stirring; (2) inoculating 1% of yeast and 2% of acetic acid bacteria by volume, culturing at a constant temperature of 35-37 ℃, stirring once every 20-24 hours, fermenting to 3-4 days, inoculating 5% of lactic acid bacteria, culturing at a constant temperature of 30 ℃ in a closed manner, and fermenting to 15 days.
In one embodiment of the present invention, the concentration of the yeast, acetic acid bacteria or lactic acid bacteria is 1 to 9.9 × 107CFU/mL。
In one embodiment of the invention, the yeast is Saccharomyces cerevisiae N85.
In one embodiment of the present invention, the acetic acid bacterium is acetobacter gluconicum (Gluconacetobacter xylinus) Q1.
In one embodiment of the invention, the lactic acid bacterium is Lactobacillus delbrueckii subsp. bulgaricus strain B1 or Lactobacillus delbrueckii subsp. bulgaricus strain B2.
In one embodiment of the present invention, the steps are specifically as follows: (1) cleaning dragon fruits, apples, tomatoes, soybeans and the like serving as raw materials with hot water, air-drying, cutting into small pieces, accurately weighing a plurality of fruits and vegetables with equal mass, uniformly mixing, and putting into a fermentation container; (2) adding 12-15% of glucose in percentage by mass of the raw materials, adding 100% of water in percentage by mass of the fruits, and fully and uniformly stirring; (3) adding 1% of yeast and 2% of acetic acid bacteria, culturing at constant temperature of 37 ℃, keeping unidirectional air exhaust, and stirring once for 24 hours; the concentration of the bacterial liquid is 107CFU/mL; (4) fermenting to 4 days, adding 5% lactobacillus, and culturing at 30 deg.C under sealed condition; the concentration of the bacterial liquid is 107CFU/mL; (5) and finishing fermentation by 15 days.
Has the advantages that: according to the invention, Saccharomyces cerevisiae N85, Gluconacetobacter gluconicum Q1 and Lactobacillus delbrueckii subsp. bulgaricus strain B1 are added into the fruit and vegetable raw materials, so that the probiotic effect of the ferment is enhanced, the reducing power is improved by 43.3% compared with that of a control group, the DPPH clearance is improved from 88.25% to 93.09%, the lactic acid content is improved by 36.1%, and the acetic acid content is improved by 35.2%. Compared with a control group, the protease activity is improved by 11.9%, the content of volatile substances is increased, the ethyl acetate content is improved by 85.1%, the amyl acetate content is improved by 89.1%, so that the ferment has special fruit fragrance, the bacteriostatic ability of the ferment is improved, and bacillus subtilis, escherichia coli and staphylococcus hominis can be effectively inhibited.
Biological material preservation
A strain of Lactobacillus delbrueckii subsp. bulgaricus strain B1 has been deposited in China center for type culture Collection in 26 months 6 in 2017 with the deposition number of CCTCC M2017370 and the deposition address of Wuhan university in China.
Drawings
FIG. 1 is a standard curve of tyrosine concentration.
Detailed Description
In the detailed description, Saccharomyces cerevisiae N85 for yellow wine industry was disclosed in 2016, entitled "research on Low-yield Ethyl carbamate by Yeast for yellow wine brewing by Metabolic engineering". Gluconacetobacter (Gluconacetobacter) Q1, which is preserved in China center for type culture Collection (CCTCCM 2014353) at 7-23.2014; lactobacillus delbrueckii subsp. bulgaricus strain B1 is screened from cheese, and has been deposited in China center for type culture Collection in 26 months 6 and 2017 with the deposit number of CCTCCM 2017370; lactobacillus delbrueckii subsp. bulgaricus strain B2 was deposited in the resource library of lactic acid bacteria strains (LABCC) in the department of dairy biotechnology and engineering education of the university of inner Mongolia agriculture, and the deposit number is LABCIMAU 20094 DBTE ML 12-2-2.
TABLE 1 culture conditions of cells according to the embodiments
And (3) organic acid determination: taking enzyme solution to be detected, diluting the enzyme solution by using ultrapure water for ten times, sucking the liquid by using a syringe, passing through an organic acid filter membrane, and injecting the liquid into a liquid phase bottle. And calibrated using mixed acid standards. The types and amounts of organic acids were measured using an Agilent 1200 hplc.
Determination of DPPH radical scavenging Capacity: diluting the enzyme sample by 5 times, adding 2mL of the enzyme sample into 2mL of 0.1mg/mL DPPH-ethanol solution, fully and uniformly mixing, carrying out dark reaction at 25 ℃ for 30min, determining the absorbance value under the condition of 517nm by taking deionized water as a reference solution, and replacing the blank group with 2mL of absolute ethanol.
DPPH clearance (/%) - [1- (a)% ]1-A2)/A0]×100
In the formula: a. the0Absorbance of blank group, A1Absorbance of the sample solution, A2Absorbance of the bottom tube is the sample.
Determination of reducing power: the measurement was carried out by potassium ferricyanide reduction. Diluting an enzyme sample to be detected by 5 times, taking 1mL of diluted sample, adding the diluted sample into a test tube filled with 1mL of phosphate buffer (0.2mol/L, pH6.6), adding 2mL of potassium ferricyanide with the mass fraction of 1%, placing the test tube in a water bath kettle at 50 ℃ for accurate reaction for 30min, adding 2mL of trichloroacetic acid with the mass fraction of 10%, centrifuging the test tube for 10min at 3000r/min, immediately taking 2mL of supernatant, adding 1mL of ferric trichloride with the mass fraction of 0.5% and 1mL of deionized water, standing for 5min, determining the absorbance value under the condition of 700nm, wherein the larger the absorbance value is, the stronger the reducing force is indicated, and the same determination is repeated twice.
Determination of protease activity: determination of tyrosine standard curves: 1mL of tyrosine solution of 0, 20, 40, 60, 80 and 100ug/mL is sucked in 6 test tubes respectively, 5mL of sodium carbonate solution of 0.4mol/L is added respectively, and 1mL of 3 times diluted forlin reagent is added. Shaking, placing in 40 deg.C water bath kettle, maintaining the temperature, developing color for 20min, and measuring absorbance at wavelength of 660 nm. The measurement was carried out three times, and the average value was taken. And drawing a standard curve by taking the absorbance as a vertical coordinate and the tyrosine concentration as a horizontal coordinate. Adding 1mL of enzyme solution into a test tube, placing the test tube in a water bath kettle at 40 ℃ for preheating for 5 minutes, then adding 1mL of 2% casein solution, precisely timing the reaction for 10 minutes, immediately adding 2mL of 0.4mol/L trichloroacetic acid solution, filtering the solution by using filter paper after 15 minutes, taking 1mL of filtrate, adding 5mL of 0.4mol/L sodium carbonate solution and 1mL of Folin reagent, preserving the temperature at 40 ℃ for 20 minutes, and measuring the optical density value at the wavelength of 660 nm. Conversion to tyrosine concentration was done according to a standard curve. Protease activity unit definition: the amount of enzyme that hydrolyzes casein at 40 ℃ and pH3.6 to release 1 microgram tyrosine per minute is defined as 1 protease unit.
In the formula:
x is the tyrosine concentration in the reaction solution;
v is the total volume (mL) of the reaction solution;
t is reaction time (min);
n is the dilution multiple of enzyme solution;
w is the volume of enzyme solution (mL).
Determination of bacteriostatic ability: the measurement was carried out by the Oxford cup method. And heating the sterilized agar culture medium to be completely melted, pouring the agar culture medium into a culture dish, wherein the volume of each dish is 20mL, and placing 4-6 oxford cups on the surface of each dish of the culture medium after solidification. At the same time, the sterilized semi-solid medium (seed medium) was heated to completely melt and dispensed into sterilized 50mL centrifuge tubes, approximately 20mL per tube. Cooling the semi-solid culture medium to about 45 deg.C, and inoculating with 10 dilution times3、104、105、106After being uniformly mixed, 100uL of the bacterial liquid is carefully poured into a culture dish with an Oxford cup, and the situation that the culture medium is poured into the Oxford cup is avoided. After the culture medium is solidified, the oxford cup is carefully taken out, round empty rings are formed on a culture dish, 100uL of a sample to be detected or a control sample is added into the empty rings, and standing culture is carried out for 16-18 h at 37 ℃.
Volatile matter determination: the measurement was carried out by GC-MS.
Example 1
1. Preparing a ferment raw material: cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding glucose accounting for 15% of the mass of the vegetable and fruit raw materials, adding water accounting for 100% of the mass of the vegetable and fruit raw materials, and fully and uniformly stirring.
2. Inoculating a strain:
activating the strain: taking out Saccharomyces cerevisiae N85 and Acetobacter gluconicum Q1 from a refrigerator at-80 ℃, and streaking and inoculating the strains in corresponding culture media respectively.
Obtaining thalli: picking single colonies on solid mediaInoculating into liquid culture medium, culturing to logarithmic phase, centrifuging at 10000rpm for 10min, discarding supernatant, and mixing with sterile physiological saline: resuspending the bacterial liquid at the ratio of 1:1 to make the bacterial concentration 1-5 multiplied by 107CFU/mL。
Inoculation: adding 1mL of bacterial liquid/100 g of fruit and vegetable-quality saccharomyces cerevisiae N85 and 2mL of bacterial liquid/100 g of fruit and vegetable-quality gluconacetobacter Q1 according to the volume, placing the mixture in a constant temperature of 37 ℃ for closed fermentation, and stirring once every 24 hours.
3. Obtaining enzyme liquid: and finishing fermentation by 15 days, centrifuging at 10000r/min, and taking supernatant as enzyme solution.
Example 2
1. Preparing a ferment raw material: cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and fully and uniformly stirring.
2. Inoculating a strain:
activating the strain: saccharomyces cerevisiae N85 and Lactobacillus delbrueckii subsp bulgaricus B1 were removed from a freezer at-80 ℃ and streaked onto the appropriate medium.
Obtaining thalli: and (3) selecting a single colony on the solid culture medium, inoculating the single colony on the solid culture medium into a liquid culture medium, respectively culturing to a logarithmic phase, centrifuging at 10000rpm for 10min, discarding supernatant, and treating the supernatant with sterile physiological saline: resuspending the bacterial liquid at the ratio of 1:1 to make the bacterial concentration 1-5 multiplied by 107CFU/mL。
Inoculation: adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 with the mass of fruits and vegetables, culturing at the constant temperature of 37 ℃, keeping unidirectional air exhaust, and stirring once for 24 hours. And (3) fermenting to the 4 th day, adding 5mL of bacterial liquid per 100g of lactobacillus delbrueckii subspecies bulgaricus B1 of fruit and vegetable in mass, and sealing and fermenting at the constant temperature of 30 ℃.
3. Obtaining enzyme liquid: and finishing fermentation by 15 days, centrifuging at 10000r/min, and taking supernatant as enzyme solution.
Example 3.
1. Preparing a ferment raw material: cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and fully and uniformly stirring.
2. Inoculating a strain:
activating the strain: saccharomyces cerevisiae N85 and Lactobacillus delbrueckii subsp bulgaricus B2 were removed from a freezer at-80 ℃ and streaked onto the appropriate medium.
Obtaining thalli: and (3) selecting a single colony on the solid culture medium, inoculating the single colony on the solid culture medium into a liquid culture medium, respectively culturing to a logarithmic phase, centrifuging at 10000rpm for 10min, discarding supernatant, and treating the supernatant with sterile physiological saline: resuspending the bacterial liquid at the ratio of 1:1 to make the bacterial concentration 1-5 multiplied by 107CFU/mL。
Inoculation: adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 with the mass of fruits and vegetables, culturing at the constant temperature of 37 ℃, keeping unidirectional air exhaust, and stirring once for 24 hours. And (3) fermenting to the 4 th day, adding 5mL of bacterial liquid per 100g of lactobacillus delbrueckii subspecies bulgaricus B2 of fruit and vegetable in mass, and sealing and fermenting at the constant temperature of 30 ℃.
3. Obtaining enzyme liquid: and finishing fermentation by 15 days, centrifuging at 10000r/min, and taking supernatant as enzyme solution.
Example 4.
1. Preparing a ferment raw material: cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and fully and uniformly stirring.
2. Inoculating a strain:
activating the strain: saccharomyces cerevisiae N85, Acetobacter gluconicum Q1 and Lactobacillus delbrueckii subsp.bulgaricus B1 were taken out of a refrigerator at-80 ℃ and streaked on the respective media.
Obtaining thalli: selecting single colony on solid culture medium, inoculating to liquid culture medium, culturing to logarithmic phase, centrifuging at 10000rpm for 10min, discarding supernatant, and sterilizingSaline as saline: resuspending the bacterial liquid at the ratio of 1:1 to make the bacterial concentration 1-5 multiplied by 107CFU/mL。
Inoculation: adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 and 2mL of bacterial liquid/100 g of acetobacter gluconicum Q1, culturing at a constant temperature of 37 ℃, keeping unidirectional air exhaust, and stirring once for 24 hours. And (3) fermenting to the 4 th day, adding 5mL of bacterial liquid per 100g of lactobacillus delbrueckii subspecies bulgaricus B1 of fruit and vegetable in mass, and sealing and fermenting at the constant temperature of 30 ℃.
3. Obtaining enzyme liquid: and finishing fermentation by 15 days, centrifuging at 10000r/min, and taking supernatant as enzyme solution.
Example 5.
1. Preparing a ferment raw material: cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and fully and uniformly stirring.
2. Inoculating a strain:
activating the strain: saccharomyces cerevisiae N85, Acetobacter gluconicum Q1 and Lactobacillus delbrueckii subsp.bulgaricus B2 were taken out of a refrigerator at-80 ℃ and streaked on the respective media.
Obtaining thalli: and (3) selecting a single colony on the solid culture medium, inoculating the single colony on the solid culture medium into a liquid culture medium, respectively culturing to a logarithmic phase, centrifuging at 10000rpm for 10min, discarding supernatant, and treating the supernatant with sterile physiological saline: resuspending the bacterial liquid at the ratio of 1:1 to make the bacterial concentration 1-5 multiplied by 107CFU/mL。
Inoculation: adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 and 2mL of bacterial liquid/100 g of acetobacter gluconicum Q1, culturing at a constant temperature of 37 ℃, keeping unidirectional air exhaust, and stirring once for 24 hours. And (3) fermenting to the 4 th day, adding 5mL of bacterial liquid per 100g of lactobacillus delbrueckii subspecies bulgaricus B2 of fruit and vegetable in mass, and sealing and fermenting at the constant temperature of 30 ℃.
3. Obtaining enzyme liquid: and finishing fermentation by 15 days, centrifuging at 10000r/min, and taking supernatant as enzyme solution.
Comparative example
1. Preparing a ferment raw material: cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and fully and uniformly stirring.
2. Inoculating a strain:
activating the strain: the Saccharomyces cerevisiae N85 was taken out from the-80 ℃ freezer and streaked on the corresponding medium.
Obtaining thalli: and (3) selecting a single colony on the solid culture medium, inoculating the single colony on the solid culture medium into a liquid culture medium, respectively culturing to a logarithmic phase, centrifuging at 10000rpm for 10min, discarding supernatant, and treating the supernatant with sterile physiological saline: resuspending the bacterial liquid at the ratio of 1:1 to make the bacterial concentration 1-5 multiplied by 107CFU/mL。
Inoculation: adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 with the mass of fruits and vegetables, placing at the constant temperature of 37 ℃ for closed fermentation, keeping unidirectional air exhaust, and stirring once for 24 hours.
3. Obtaining enzyme liquid: and finishing fermentation by 15 days, centrifuging at 10000r/min, and taking supernatant as enzyme solution.
The enzymes prepared in examples 1 to 5 and the control example were subjected to measurement of oxidation resistance, organic acid content, protease activity, bacteriostatic ability, and volatile substance.
The results of the antioxidant capacity measurement are shown in Table 2, the enzyme enhanced by Lactobacillus delbrueckii subsp-bulgaricus B1 has improved antioxidant capacity, wherein the reducing power is improved by 43.3%, the DPPH clearance is improved from 88.25% to 93.09%, and the enzyme reducing power and DPPH clearance are only 21.2% and 91.07% when the Lactobacillus delbrueckii subsp-bulgaricus B2 is added.
TABLE 2 antioxidative Activity of enzymes in various embodiments
The measurement results of the organic acid content are shown in table 3, the enzyme reinforced by lactobacillus delbrueckii subsp bulgaricus B1 has the organic acid content improved compared with that before the enzyme is not added, wherein the lactic acid content is improved by 36.1%, and the acetic acid content is improved by 35.2%; the acetic acid and lactic acid contents of the ferment added to lactobacillus delbrueckii subsp bulgaricus B2 appeared to be reduced, which was detrimental to the storage and quality of the product.
TABLE 3 organic acid content (g/L) of ferment in different embodiments
The protease activity determination results are shown in table 4, the tyrosine standard curve is shown in fig. 1, and the protease activity of the enzyme enhanced by lactobacillus delbrueckii subspecies bulgaricus B1 is improved by 11.9%.
TABLE 4 protease Activity of enzymes according to various embodiments
The results of the bacteriostatic ability measurement are shown in table 5, the enzyme reinforced by lactobacillus delbrueckii subsp bulgaricus B1 shows the inhibitory action on bacillus subtilis, escherichia coli and human staphylococcus, and the diameters of the bacteriostatic circle are 15.3mm, 16.6mm and 14.3mm respectively; the enzyme added with Lactobacillus delbrueckii subspecies B2 has no bacteriostatic ability.
TABLE 5 bacteriostatic ability of the enzyme in different embodiments (d/mm)
The measurement results of volatile substances are shown in table 6, the ethanol content of the ferment enhanced by the lactobacillus delbrueckii subsp bulgaricus B1 in the volatile substances is reduced by 20.1%, the ferment is suitable for more people to eat, the ethyl acetate content is increased by 85.1%, the amyl acetate content is increased by 89.1%, and the fruit fragrance is increased; compared with Lactobacillus delbrueckii subspecies bulgaricus B1, the ethanol content of the ferment added with the Lactobacillus delbrueckii subspecies bulgaricus is increased by 6.0%, the ethyl acetate content is reduced by 70.9%, and the flavor is not good.
TABLE 6 volatile substance content (g/L) of ferments in different embodiments
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (5)
1. A method for improving the functionality and flavor of fruit and vegetable ferment simultaneously is characterized in that pitaya, apple, tomato and soybean are cleaned by hot water, dried in air on a clean workbench and cut into small blocks; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and uniformly stirring; taking out Saccharomyces cerevisiae N85, gluconacetobacter Q1 and Lactobacillus delbrueckii subsp bulgaricus B1 from a refrigerator at the temperature of-80 ℃, streaking and inoculating the materials to corresponding culture media, selecting a single colony on a solid culture medium to be inoculated to a liquid culture medium, respectively culturing the single colony to a logarithmic growth phase, centrifuging the single colony at 10000rpm for 10min, discarding supernatant, and treating the supernatant with sterile physiological saline: resuspending the bacterial liquid at a ratio of 1:1 to make the concentration of the bacterial liquid 1-5 × 107 CFU/mL; adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 and 2mL of bacterial liquid/100 g of acetobacter gluconicum Q1, culturing at a constant temperature of 37 ℃, keeping unidirectional exhaust, stirring once for 24 hours, fermenting to 4 days, adding 5mL of bacterial liquid/100 g of lactobacillus delbrueckii subspecies bulgaricus B1, and performing closed fermentation at a constant temperature of 30 ℃; after the fermentation is finished by 15 days, centrifuging at 10000r/min to obtain a supernatant as enzyme solution; the Lactobacillus delbrueckii subspecies bulgaricus (B.delbrueckii)Lactobacillus delbrueckiisubsp.bulgaricus strain) B1, which has been 26 in 2017 month 6The product is preserved in China center for type culture Collection with preservation number of CCTCC NO: M2017370, and the preservation address of Wuhan, Wuhan university.
2. A ferment is characterized in that the ferment is prepared by fermenting dragon fruit, apple, tomato and soybean with microorganism, and the thallus number is more than or equal to 1 multiplied by 108 Yeast, gluconacetobacter and lactobacillus; the yeast is Saccharomyces cerevisiae N85, the gluconacetobacter is gluconacetobacter Q1, and the lactobacillus is Lactobacillus delbrueckii subspecies B1; the ferment is prepared by the following steps: cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and uniformly stirring; taking out Saccharomyces cerevisiae N85, gluconacetobacter Q1 and Lactobacillus delbrueckii subsp bulgaricus B1 from a refrigerator at the temperature of-80 ℃, streaking and inoculating the materials to corresponding culture media, selecting a single colony on a solid culture medium to be inoculated to a liquid culture medium, respectively culturing the single colony to a logarithmic growth phase, centrifuging the single colony at 10000rpm for 10min, discarding supernatant, and treating the supernatant with sterile physiological saline: resuspending the bacterial liquid at a ratio of 1:1 to make the concentration of the bacterial liquid 1-5 × 107 CFU/mL; adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 and 2mL of bacterial liquid/100 g of acetobacter gluconicum Q1, culturing at a constant temperature of 37 ℃, keeping unidirectional exhaust, stirring once for 24 hours, fermenting to 4 days, adding 5mL of bacterial liquid/100 g of lactobacillus delbrueckii subspecies bulgaricus B1, and performing closed fermentation at a constant temperature of 30 ℃; after the fermentation is finished by 15 days, centrifuging at 10000r/min to obtain a supernatant as enzyme solution; the lactobacillus is Lactobacillus delbrueckii subspecies bulgaricus (B.delbrueckii)Lactobacillus delbrueckiisubsp.bulgaricus strain) B1, which has been preserved in China Center for Type Culture Collection (CCTCC) No. M2017370 in 2017, 6.26.7, and the preservation address is Wuhan, Wuhan university.
3. A preparation method of ferment is characterized by comprising the following steps:cleaning dragon fruit, apple, tomato and soybean with hot water, air drying on a clean workbench, and cutting into small pieces; weighing 100g of each fruit and vegetable, uniformly mixing, putting into a fermentation container, adding 15% by mass of glucose as a raw material of the fruit and vegetable, adding 100% by mass of water as the raw material of the fruit and vegetable, and uniformly stirring; taking out Saccharomyces cerevisiae N85, gluconacetobacter Q1 and Lactobacillus delbrueckii subsp bulgaricus B1 from a refrigerator at the temperature of-80 ℃, streaking and inoculating the materials to corresponding culture media, selecting a single colony on a solid culture medium to be inoculated to a liquid culture medium, respectively culturing the single colony to a logarithmic growth phase, centrifuging the single colony at 10000rpm for 10min, discarding supernatant, and treating the supernatant with sterile physiological saline: resuspending the bacterial liquid at a ratio of 1:1 to make the concentration of the bacterial liquid 1-5 × 107 CFU/mL; adding 1mL of bacterial liquid/100 g of saccharomyces cerevisiae N85 and 2mL of bacterial liquid/100 g of acetobacter gluconicum Q1, culturing at a constant temperature of 37 ℃, keeping unidirectional exhaust, stirring once for 24 hours, fermenting to 4 days, adding 5mL of bacterial liquid/100 g of lactobacillus delbrueckii subspecies bulgaricus B1, and performing closed fermentation at a constant temperature of 30 ℃; after the fermentation is finished by 15 days, centrifuging at 10000r/min to obtain a supernatant as enzyme solution; the Lactobacillus delbrueckii subspecies bulgaricus (B.delbrueckii)Lactobacillus delbrueckiisubsp.bulgaricus strain) B1, which has been preserved in China Center for Type Culture Collection (CCTCC) No. M2017370 in 2017, 6.26.7, and the preservation address is Wuhan, Wuhan university.
4. Food product comprising an extract or concentrate of the ferment of claim 2.
5. Lactobacillus delbrueckii subspecies bulgaricus (L.) (Lactobacillus delbrueckiisubsp.bulgaricus strain) B1, which is preserved in China Center for Type Culture Collection (CCTCC) No. M2017370 in 26.6.2017, with the preservation address of Wuhan, Wuhan university.
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| CN108740014B (en) * | 2018-06-15 | 2021-09-07 | 光明乳业股份有限公司 | High-fruity yoghourt and preparation method thereof |
| CN109288049A (en) * | 2018-11-23 | 2019-02-01 | 湖北尧生物科技有限公司 | A kind of preparation method of Stropharia rugoso-annulata biologic ferment |
| CN109757718A (en) * | 2019-02-19 | 2019-05-17 | 新疆农业科学院微生物应用研究所（中国新疆-亚美尼亚生物工程研究开发中心） | A kind of delicate fragrance type Xinjiang little white apricot ferment and preparation method thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102940288A (en) * | 2012-11-19 | 2013-02-27 | 商丘市饮之健食品有限公司 | Composite enzyme beverage and preparation process method thereof |
| KR101485896B1 (en) * | 2014-10-27 | 2015-01-26 | 주식회사 더트루메틱 | Cosmetic composition for improving anti-oxidation, anti-inflammatory and atopic skin and method of preparing the same |
| CN104328073A (en) * | 2014-10-30 | 2015-02-04 | 江南大学 | Gluconacetobacter xylinus strain capable of producing free glucuronic acid |
| CN104886572A (en) * | 2015-06-02 | 2015-09-09 | 广州加海生物科技有限公司 | A fruit and vegetable ferment beverage and a preparation method thereof |
| CN105533544A (en) * | 2015-12-31 | 2016-05-04 | 天津天绿健科技有限公司 | Natural fruit and vegetable enzyme product |
| CN105614858A (en) * | 2015-12-31 | 2016-06-01 | 天津天绿健科技有限公司 | Plant ferment product and preparation thereof |
| CN105639525A (en) * | 2016-02-03 | 2016-06-08 | 大管家（上海）健康产业有限公司 | Fruit and vegetable enzyme capable of protecting liver and expelling toxins and preparing method of fruit and vegetable enzyme |
| CN106213095A (en) * | 2016-06-30 | 2016-12-14 | 恒枫食品科技有限公司 | The preparation method of a kind of fermented fruits and vegetables juice and obtain the application of product |
| CN106343547A (en) * | 2016-08-22 | 2017-01-25 | 宁波希诺亚海洋生物科技有限公司 | Preparation method for plant/fruit/vegetable enzymes |
| CN106578805A (en) * | 2016-06-07 | 2017-04-26 | 上海融扬生物技术有限公司 | Functional fruit and vegetable enzyme beverage |
-
2017
- 2017-10-18 CN CN201710972121.6A patent/CN107788517B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102940288A (en) * | 2012-11-19 | 2013-02-27 | 商丘市饮之健食品有限公司 | Composite enzyme beverage and preparation process method thereof |
| KR101485896B1 (en) * | 2014-10-27 | 2015-01-26 | 주식회사 더트루메틱 | Cosmetic composition for improving anti-oxidation, anti-inflammatory and atopic skin and method of preparing the same |
| CN104328073A (en) * | 2014-10-30 | 2015-02-04 | 江南大学 | Gluconacetobacter xylinus strain capable of producing free glucuronic acid |
| CN104886572A (en) * | 2015-06-02 | 2015-09-09 | 广州加海生物科技有限公司 | A fruit and vegetable ferment beverage and a preparation method thereof |
| CN105533544A (en) * | 2015-12-31 | 2016-05-04 | 天津天绿健科技有限公司 | Natural fruit and vegetable enzyme product |
| CN105614858A (en) * | 2015-12-31 | 2016-06-01 | 天津天绿健科技有限公司 | Plant ferment product and preparation thereof |
| CN105639525A (en) * | 2016-02-03 | 2016-06-08 | 大管家（上海）健康产业有限公司 | Fruit and vegetable enzyme capable of protecting liver and expelling toxins and preparing method of fruit and vegetable enzyme |
| CN106578805A (en) * | 2016-06-07 | 2017-04-26 | 上海融扬生物技术有限公司 | Functional fruit and vegetable enzyme beverage |
| CN106213095A (en) * | 2016-06-30 | 2016-12-14 | 恒枫食品科技有限公司 | The preparation method of a kind of fermented fruits and vegetables juice and obtain the application of product |
| CN106343547A (en) * | 2016-08-22 | 2017-01-25 | 宁波希诺亚海洋生物科技有限公司 | Preparation method for plant/fruit/vegetable enzymes |
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