CN110004090B - Leuconostoc mesenteroides and application thereof in pickled vegetable fermentation - Google Patents
Leuconostoc mesenteroides and application thereof in pickled vegetable fermentation Download PDFInfo
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- 229910002651 NO3 Inorganic materials 0.000 abstract description 5
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 abstract description 5
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- 238000012360 testing method Methods 0.000 description 16
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B7/00—Preservation or chemical ripening of fruit or vegetables
- A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
- A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
- A23B7/154—Organic compounds; Microorganisms; Enzymes
- A23B7/155—Microorganisms; Enzymes; Antibiotics
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- 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
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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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
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
- A23L5/28—Removal of unwanted matter, e.g. deodorisation or detoxification using microorganisms
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- 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
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/31—Leuconostoc
- A23V2400/321—Mesenteroides
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
The invention discloses Leuconostoc mesenteroides and application thereof in pickled vegetable fermentation, and relates to the technical field of fermented foods, wherein the Leuconostoc mesenteroides DN-LM-D2 comprises the following steps of: 1) selecting fresh Chinese cabbage, and performing pretreatment; 2) cleaning: washing the Chinese cabbages treated in the step 1) with water, draining, and placing in a fermentation container; 3) fermentation: and (3) injecting purified water into the fermentation container filled with the Chinese cabbages, then adding leuconostoc mesenteroides liquid, adding edible salt at the same time, and performing closed fermentation to obtain the pickled Chinese cabbages. The method provided by the invention has the advantages of low nitrate and nitrite content and high dish yield when used for fermenting the pickled Chinese cabbage.
Description
Technical Field
The invention relates to the technical field of fermented foods, in particular to leuconostoc mesenteroides and application thereof in fermented pickled Chinese cabbage.
Background
The pickled Chinese cabbage is a traditional fermented vegetable product in the north of China, has unique flavor and is tasty and refreshing, and the appetite can be improved. The pickled Chinese cabbage fermentation liquor is rich in lactic acid bacteria, and aromatic substances generated in the growth and metabolism process of the pickled Chinese cabbage fermentation liquor make the pickled Chinese cabbage unique in flavor. The main metabolites of lactic acid bacteria include acid metabolites, exopolysaccharides, lactobacillin, gamma-aminobutyric acid and the like, and the substances play roles in regulating body immunity, resisting tumors, resisting bacteria and preventing corrosion. In addition, the pickled Chinese cabbage contains rich nutrients such as vitamins, minerals, dietary fibers and the like, and is a food with the effects of reducing cholesterol, regulating intestines and stomach and expelling toxin. The fermented pickled Chinese cabbage is not only a processed food, but also can prolong the storage period of the Chinese cabbage, and is a nutritional and effective Chinese cabbage processing means. The method is popular with residents in China, particularly in the northeast, and has great market potential.
The fermentation mode of the pickled Chinese cabbage is divided into two modes of natural fermentation and artificial inoculation fermentation. The pickled Chinese cabbages pickled by the traditional natural fermentation method are mellow in taste and rich in smell, but are long in time consumption, unstable in product quality, easy to rot and smell, and easy to generate substances such as nitrite harmful to human bodies. While the industrial fermentation has short time consumption and uniform product quality, the taste of the industrial fermentation is far inferior to the natural fermentation of the pickled Chinese cabbage, and the flavor is mellow and full.
Disclosure of Invention
In order to solve the problems, the invention provides leuconostoc mesenteroides and application thereof in pickled vegetable fermentation.
The Leuconostoc mesenteroides is named as Leuconostoc mesenteroides DN-LM-D2, classified as Leuconostoc mesenteroides, and has the laboratory number of DN-LM-D2, the preservation number of CGMCC No.16442, the preservation date of 2018, 9 and 10 days, and the preservation unit address of No. 3 Homew No.1 of Beijing Korean district.
The colony of the Leuconostoc mesenteroides DN-LM-D2 is white opaque circle, gram staining is positive, thallus is spherical, and the diameter is less than or equal to 1 mm. The test results of catalase test, starch hydrolysis test, arginine ammonia production test, gas production test and motility test are negative, and the test results of methyl red test, acid production reaction and oxidase test are positive.
Secondly, the invention also provides application of the leuconostoc mesenteroides fermented pickled Chinese cabbage.
The application comprises the following steps:
1) selecting fresh Chinese cabbage, and performing pretreatment;
2) cleaning: washing the Chinese cabbages treated in the step 1) with water, draining, and placing in a fermentation container;
3) fermentation: and (3) injecting purified water into the fermentation container filled with the Chinese cabbages, then adding leuconostoc mesenteroides liquid, adding edible salt at the same time, and performing closed fermentation to obtain the pickled Chinese cabbages.
The pretreatment of the step 1) is specifically to strip off the rotten or dehydrated part of the outer layer and place the rotten or dehydrated part in a cool, dry and ventilated place.
The standing time is 2-3 days.
And 3) injecting sterile water in an amount that the liquid level of the sterile water is over the Chinese cabbage.
Step 3) the Leuconostoc mesenteroides is Leuconostoc mesenteroides DN-LM-D2.
Step 3) the bacteria concentration of the leuconostoc mesenteroides liquid is 1 multiplied by 107-1.8×107cfu/ml。
And 3) adding the leuconostoc mesenteroides bacterial liquid into the Chinese cabbage in an amount which is 4% of the mass fraction of the Chinese cabbage obtained after the pretreatment in the step 1).
And 3) adding the edible salt in an amount which is 2% of the mass fraction of the Chinese cabbage obtained after the pretreatment in the step 1).
And 3) performing closed fermentation, wherein the fermentation temperature is 20 ℃, and the fermentation time is more than 30 days.
The preparation method of the leuconostoc mesenteroides bacterial liquid in the step 3) comprises the following steps: streaking a Leuconostoc mesenteroides DN-LM-D2 strain liquid on an MRS plate, culturing for 48-72h at 37 ℃, selecting a single colony, inoculating the single colony into an MRS liquid culture medium, performing shake culture for 18-24h at 37 ℃ at 150rps to serve as a seed liquid, inoculating the seed liquid into an amplification culture medium according to the volume ratio of 5-20%, and performing amplification culture to obtain the Leuconostoc mesenteroides liquid.
The formula of the amplification culture medium comprises 10g/L of peptone, 5g/L of yeast powder, 5g/L of beef extract, 100ml/L of tomato juice and the balance of water, and the pH value of the culture medium is adjusted to 6.2-6.4 by using 1M NaOH. The expanding culture is carried out under the conditions of 37 ℃ and 150rps shaking culture, and the expanding culture time is 18-24 h.
Advantageous effects
Leuconostoc mesenteroides DN-LM-D2 strain has strong capability of degrading nitrite, reduces nitrite content in fermentation liquor, reduces hidden danger of eating and improves edible safety of vegetable fermentation products, and the content of nitrate and nitrite in pickled vegetables fermented by the strain provided by the invention is respectively reduced by 31.9% and 38.5% compared with that of pickled vegetables naturally fermented under the same condition. Meanwhile, Leuconostoc mesenteroides (Leuconostoc mesenteroides) DN-LM-D2 has stronger acid resistance, which is beneficial to maintaining acid production of fermentation liquor in the later period of fermentation and improving the accumulation of flavor substances, the taste is closer to that of the traditional fermented pickled vegetables, and the pH is basically consistent with that of the naturally fermented pickled vegetables. In addition, due to the artificial inoculation of Leuconostoc mesenteroides DN-LM-D2 bacteria, the Leuconostoc mesenteroides becomes a dominant flora in the fermentation process, the growth of mixed bacteria and harmful bacteria is inhibited, the dish yield (the quality of pickled cabbage which has enough acidity and is not rotten after the fermentation is finished/the quality of initial Chinese cabbage) is increased, and the quality of the fermented vegetable products is improved, and the dish yield is 22% higher than that of the pickled cabbage which is naturally fermented under the same condition by utilizing the strain provided by the invention to ferment the pickled cabbage.
Detailed Description
EXAMPLE 1 screening and identification of bacterial species
Separating and purifying strains
Selecting a little of traditional fermented pickled Chinese cabbage self-made by a plurality of citizens in the Harbin region, alternately washing the surfaces of the pickled Chinese cabbage by sterile water and 70% ethanol for 3 times, washing the pickled Chinese cabbage by the sterile water, and draining the redundant water. Grinding pickled Chinese cabbage leaf to obtain juice, and diluting with sterile water in gradient manner (10)-1、10-2、10-3、10-4、10-5) Taking 50 microliters of each diluted concentration culture solution, coating the diluted concentration culture solution on an MRS solid culture medium, culturing for 48 hours at a constant temperature of 37 ℃, and selecting a single colony with a larger calcium-dissolving ring to streak and purify on the MRS solid culture medium. The 11 colonies which generate the calcium-dissolving rings are screened out and named as DN-LP-X1, DN-LP-X2, DN-LP-M1, DN-LP-M2, DN-LP-W1, DN-LP-H2, DN-LA-H7, DN-LM-Y2, DN-LM-Y3, DN-LM-Y4 and DN-LM-D2 respectively. The 11 strains were cultured in MRS liquid medium at 37 ℃ for 12h at 150rpm/min with 60% glycerol in a ratio of 1: 1, and placing the mixture into a sterile tube to be stored at the temperature of minus 80 ℃.
Physiological and biochemical and strain molecular identification
Morphological characterization observation shows that all the 11 colonies are white opaque circles and gram staining is positive. Wherein 9 strains of bacteria such as DN-LP-X1, DN-LP-X2, DN-LP-M1, DN-LP-M2, DN-LP-W1, DN-LP-H2, DN-LA-H7 and the like are rod-shaped, the diameter is 1-2mm, four strains of bacteria such as DN-LM-Y2, DN-LM-Y3, DN-LM-Y4, DN-LM-D2 and the like are spherical, and the diameter is less than or equal to 1 mm. Then, physiological and biochemical characteristics are identified to show that the catalase test, the starch hydrolysis test, the arginine ammonia production test, the gas production test and the motility test of the 11 strains are negative, the methyl red test, the acid production reaction and the oxidase test are positive, and the 11 strains are initially identified as the lactobacillus.
And (3) identifying the strain molecules: 16S rDNA universal primer is utilized, the bacterial liquid genome is taken as a template for PCR amplification, and the amplified product is subjected to agarose electrophoresis and purification and then sent to Beijing Huada Biometrics, Inc. for sequencing. The results of alignment and sequencing by using NCBI database find that the sequence similarity of DN-LP-X1, DN-LP-X2, DN-LP-M1, DN-LP-M2, DN-LP-W1, DN-LP-H2 and DN-LA-H7 with Lactobacillus plantarum is up to 100 percent, and the strains are lactobacillus plantarum. And the similarity of DN-LM-Y3 and DN-LM-Y4 and Leuconostoc mesenteroides is up to 100 percent, and the similarity of DN-LM-Y2 and DN-LM-D2 and Leuconostoc mesenteroides is up to 99 percent, which indicates that DN-LM-Y2, DN-LM-Y3, DN-LM-Y4 and DN-LM-D2 are Leuconostoc mesenteroides.
And (3) identifying the nitrite degrading and acid resisting capability of the strain.
Nitrite degradation capacity identification
All strains were activated to 11.8X 107-1.8×107cfu/ml (11 bacteria adjusted to the same bacteria concentration) and inoculated in a solution containing 10mg/L NaNO at 1% by volume2In MRS liquid culture medium, culturing at 37 ℃ for 24h, and then determining NaNO2The degradation rate of (c). The strains with the strongest capacity of degrading nitrite are found to be DN-LM-Y4 and DN-LM-D2, which respectively reach 95.32 percent and 98.31 percent.
TABLE 1 nitrite degradation rate of the strains
Acid resistance assay
The most suitable pH value for the 11 lactobacillus to grow in the MRS liquid culture medium is 6; when pH 3.5, the growth of all strains was inhibited to different degrees, and the acid resistance was stronger for the strain with the least decrease in absorbance, i.e., OD600(pH 6.5) -OD600(pH 3.5) -OD 521, as compared to OD600 at pH 6, with the least decrease in absorbance of DN-LM-D2, i.e., OD600(pH 6.5) -OD600(pH 3.5) -0.521, and for the other strains DN-LP-X1, DN-LP-X2, DN-LP-M1, DN-LP-M2, LM-LP-W1, DN-LP-H2, DN-LA-H7, DN-LM-Y2, DN-LM-Y3, and DN-LM-Y4, 1.452, 1.459, 1.407, 1.369, 1.433, 1.322, 1.389, 1.183, 1.231, 0. 0.874, 0. 2, and thus better acid resistance was indicated.
Example 2 application of Leuconostoc mesenteroides DN-LM-D2 in pickling sauerkraut
Selecting a bacterial liquid of Leuconostoc mesenteroides DN-LM-D2 preserved in a glycerol tube, streaking the bacterial liquid on an MRS plate, culturing for 48-72h at 37 ℃, selecting a single colony, inoculating the single colony into 20-30ml of MRS liquid culture medium, culturing by shaking at 37 ℃ and 150r to obtain 18-serving as a seed liquid to be subjected to amplification culture. The formula of the expanded fermentation medium comprises 10g/L of peptone, 5g/L of yeast powder, 5g/L of beef extract and 100ml/L of tomato juice, the pH value of the medium is adjusted to 6.2-6.4 by 1M NaOH, and the leuconostoc mesenteroides liquid is obtained after culturing for 18 h. The bacterial liquid is used for fermenting pickled Chinese cabbage:
1) selecting fresh Chinese cabbage, removing outer leaf (rotten or dehydrated leaf), and standing in shady, dry and ventilated place for 2-3 days.
2) Cleaning: washing Chinese cabbage with clear water, draining, and placing in a fermentation container.
3) Fermentation: injecting purified water into the fermentation container until the liquid surface does not cover the Chinese cabbage, adding Leuconostoc mesenteroides liquid according to 4% of the weight of the Chinese cabbage, and simultaneously adding edible salt according to 2% of the weight of the Chinese cabbage. Fermenting at 20 deg.C under sealed condition (30 days) to obtain sauerkraut product.
Comparative example 1 Natural fermented pickled vegetable
1) Selecting fresh Chinese cabbage, removing outer leaf (rotten or dehydrated leaf), and standing in shady, dry and ventilated place for 2-3 days.
2) Cleaning: washing Chinese cabbage with clear water, draining, and placing in a fermentation container.
3) Fermentation: injecting purified water into the fermentation container until the liquid surface does not cover the Chinese cabbage, and adding edible salt according to 2% of the weight of the Chinese cabbage. Fermenting at 20 deg.C under sealed condition (30 days) to obtain sauerkraut product.
Comparative example 2 fermentation of pickled vegetable with other lactic acid bacteria
The simultaneously screened strain Leuconostoc mesenteroides DN-LM-Y4 is used to replace the Leuconostoc mesenteroides DN-LM-D2 strain in the example 2, and the other conditions are the same as the example 2:
selecting a bacterial liquid of Leuconostoc mesenteroides DN-LM-Y4 preserved in a glycerol tube, streaking the bacterial liquid on an MRS plate, culturing for 48-72h at 37 ℃, selecting a single colony, inoculating the single colony into 20-30ml of MRS liquid culture medium, culturing for 18-24h at 37 ℃ and 150r by a shaking table to serve as a seed liquid for amplification culture. The formula of the expanded fermentation medium comprises 10g/L of peptone, 5g/L of yeast powder, 5g/L of beef extract and 100ml/L of tomato juice, the pH value of the medium is adjusted to 6.2-6.4 by 1M NaOH, and the Leuconostoc mesenteroides DN-LM-Y4 bacterial liquid is obtained after culturing for 18 h. The bacterial liquid is used for fermenting pickled Chinese cabbage:
1) selecting fresh Chinese cabbage, removing outer leaves (rotten or shriveled leaves), and standing in a cool, dry and ventilated place for 2-3 days.
2) Cleaning: washing Chinese cabbage with clear water, draining, and placing in a fermentation container.
3) Fermentation: purified water is injected into the fermentation container until the liquid level does not exceed the Chinese cabbage, and then Leuconostoc mesenteroides DN-LM-Y4 bacterial liquid accounting for 4% of the weight of the Chinese cabbage is added, and edible salt accounting for 2% of the weight of the Chinese cabbage is added at the same time. Fermenting at 20 deg.C under sealed condition (30 days) to obtain sauerkraut product.
The pH (fermentation broth), nitrate content and nitrite content of the fermented pickled Chinese cabbage are respectively measured, and the specific measurement results are shown in Table 2.
TABLE 2 DN-LM-D2 comparison of inoculated fermented pickled vegetable and naturally fermented pickled vegetable
As can be seen from the data in Table 2, compared with natural fermentation, the content of nitrate in the pickled Chinese cabbage fermented by using the strain provided by the invention is reduced by 31.9%, the content of nitrite in the pickled Chinese cabbage is reduced by 38.5%, and the pH value of the pickled Chinese cabbage fermented by using the method is slightly lower than that of the naturally fermented pickled Chinese cabbage, so that the acidity of the pickled Chinese cabbage fermented by using the method provided by the invention is higher than that of the naturally fermented pickled Chinese cabbage; in addition, as can be seen from the table above, compared with the pickled Chinese cabbage fermented by other lactic acid bacteria, the content of nitrate is reduced by 12.4%, the content of nitrite is reduced by 15.8%, the pH value of the pickled Chinese cabbage fermented by other lactic acid bacteria is higher than that of the pickled Chinese cabbage fermented by natural fermentation, the pH value is far higher than that of the pickled Chinese cabbage fermented by the method, the acidity is too low, and the taste of the pickled Chinese cabbage is seriously influenced.
The content of mixed bacteria (the number of bacteria/volume of fermentation liquor) except the lactic acid bacteria in the fermentation liquor is respectively detected, the comparative example 2 is 37 times of that in the example 2, and the comparative example 1 has the highest content of the mixed bacteria, which is 118 times of that in the example 2. The vegetable yield (the quality of pickled vegetables which have sufficient acidity and are not rotten after the fermentation is finished/the quality of the initial Chinese cabbage) is calculated respectively, and the vegetable yield of example 2 is 22% higher than that of the pickled vegetables which are naturally fermented under the same condition (comparative example 1) and 9% higher than that of the comparative example 2.
Claims (7)
1. A leuconostoc mesenteroides is characterized in that: the leuconostoc mesenteroides is named as leuconostoc mesenteroidesLeuconostoc mesenteroides DN-LM-D2, class nameLeuconostoc mesenteroidesThe biological preservative is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.16442 and the preservation date of 2018, 9 months and 10 days.
2. The application of leuconostoc mesenteroides of claim 1 in fermenting pickled Chinese cabbage, characterized in that: the method comprises the following steps:
1) selecting fresh Chinese cabbage, and performing pretreatment;
2) cleaning: washing the Chinese cabbages treated in the step 1) with water, draining, and placing in a fermentation container;
3) fermentation: injecting purified water into a fermentation container filled with Chinese cabbage, adding Leuconostoc mesenteroides liquid, adding edible salt, and fermenting under sealed condition at 20 deg.C for more than 30 days to obtain sauerkraut.
3. The use of leuconostoc mesenteroides of claim 2 in fermenting pickled Chinese cabbage, characterized in that: the pretreatment of the step 1) is specifically that the outer layer rotten or shrivelled part is stripped off, and the part is placed in a cool, dry and ventilated place for 2 to 3 days.
4. The use of leuconostoc mesenteroides of claim 2 in fermenting pickled Chinese cabbage, characterized in that: and 3) injecting the purified water in an amount that the liquid level of the purified water is over the Chinese cabbage.
5. The use of leuconostoc mesenteroides of claim 2 in fermenting pickled Chinese cabbage, characterized in that: step 3) the bacteria concentration of the leuconostoc mesenteroides liquid is 1 multiplied by 107-1.8×107 cfu/ml。
6. The use of leuconostoc mesenteroides of claim 2 in fermenting pickled Chinese cabbage, characterized in that: and 3) adding the leuconostoc mesenteroides bacterial liquid into the Chinese cabbage in an amount which is 4% of the mass fraction of the Chinese cabbage obtained after the pretreatment in the step 1).
7. The use of leuconostoc mesenteroides of claim 2 in fermenting pickled Chinese cabbage, characterized in that: and 3) adding the edible salt in an amount which is 2% of the mass fraction of the Chinese cabbage obtained after the pretreatment in the step 1).
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