CN105077113A - Pickle containing mixed lactic acid bacteria and making method thereof - Google Patents

Abstract

The invention discloses a pickle containing mixed lactic acid bacteria and a preparation method thereof, wherein the mixed lactic acid bacteria include Lactobacillus (Lactobacillus? sp.) DMDL? 9010 and Lactobacillus casei subsp. Rhamnosus (Lactobacillus? casei? subsp. Rhamnosus), said Lactobacillus has been preserved in the General Microorganism Center of China Microbiological Culture Collection Management Committee on August 19, 2011, and the preservation number is CGMCC ? No. 5172. After adding fresh vegetables to mixed lactic acid bacteria and anaerobically fermenting at 20-40°C for 60-72 hours, the number of viable lactic acid bacteria in kimchi obtained at room temperature for ⁶ months can reach 105-106 ^cfu /g, and the salt content is less than 3.0% . The fermented kimchi of the invention hardly contains nitrite, has short production cycle, stable product quality, contains live probiotics and can be eaten immediately.

Description

A kind of kimchi containing mixed lactic acid bacteria and its preparation method

technical field

The invention belongs to the technical field of food, and specifically refers to a pickle containing mixed lactic acid bacteria and a preparation method thereof.

Background technique

Probiotics are live microbial food ingredients that benefit human health. Probiotics are usually lactic acid bacteria or bifidobacteria, mainly of the Lactobacillus genus, but also include some other enterococci, propionibacterium and Clostridium.

On the one hand, because fresh vegetables contain a lot of nitrates, some harmful bacteria in pickles will be converted into nitrites under the action of nitrate reductase during the fermentation process. At the same time, phenolic substances and vitamin C in vegetables Substances such as nitrate will also have a certain reducing effect on nitrate, and after the excessive nitrite in pickles is eaten into the stomach, it will react with the secondary amine of protein decomposition products under the action of hydrochloric acid to form nitrosamines, which has a strong carcinogenic effect. Teratogenic effect. According to reports, 80% of the nitrite ingested by the human body comes from vegetables. On the other hand, since people consume more animal foods in their daily diet, it is easy to cause the cholesterol content in the human body to exceed the standard, resulting in hypercholesterolemia, coronary heart disease, atherosclerosis, stroke and other cardiovascular and cerebrovascular diseases. Therefore, it is particularly important to study the application of Lactobacillus plantarum DMDL9010, which has a strong ability to degrade nitrite and cholesterol, in fermented kimchi. In recent years, many scholars have used artificial inoculation of lactic acid bacteria to shorten the fermentation period of vegetable products, improve the quality of fermented vegetables, and significantly reduce the content of nitrite in fermented vegetables. Reports on the application of kimchi food.

The patent application number is 94111749.9. The Chinese invention patent discloses a process for producing kimchi using purebred lactic acid bacteria. It takes about 15 days to mature at room temperature, and it needs to add up to 13.5-15.0% salt. The process has a long growth cycle and contains high Level of salt, with the continuous improvement of people's living standards, the requirements for food quality are getting higher and higher. Fermented kimchi with low salinity and stronger health care functions is more beneficial to people's health, so low salt and helps to degrade the human body Cholesterol fermented kimchi has become the development direction of fermented vegetables at home and abroad.

Contents of the invention

The purpose of the present invention is to provide a pickle containing mixed lactic acid bacteria and a preparation method in order to solve the above-mentioned deficiencies in the prior art.

The purpose of the present invention is achieved through the following technical solutions.

A method for preparing pickles containing mixed lactic acid bacteria, wherein the mixed lactic acid bacteria include Lactobacillus sp. DMDL9010 and Lactobacillus caseisubsp. Rhamnosus.

A preparation method for pickles containing mixed lactic acid bacteria, comprising the steps of:

(1) Place the freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached above 10 ⁸ CFU/mL;

(2) Place the seed liquids of Lactobacillus DMDL9010 and Lactobacillus casei subspecies rhamnose in the fermentation medium at a volume percentage of 1% to 10%, and culture them statically at 30°C to 40°C for 18h to 36h, Prepare a fermentation broth so that the number of viable bacteria reaches 10 ⁸ CFU/mL or more; the formulation of the fermentation medium is: 80-90 parts of fresh vegetable juice, 0.5-3.0 parts of vitamin C, 0.5-3.0 parts of NaCl Parts, dilute to 100 parts with sterile water, after sterilization and cooling, it is the fermentation medium;

(3) In parts by weight, 20 to 40 parts of vegetables are inserted into 3 to 10 parts of the fermentation broth of Lactobacillus DMDL9010 and Lactobacillus casei subsp. Add 0.1 to 2.0 parts of vitamin C and 0.2 to 5.0 parts of NaCl each, dilute to 100 parts with sterile water, and immerse the vegetables in the liquid;

(4) Carry out anaerobic fermentation under sealed conditions, and ferment for 24-72 hours at a temperature of 20-40°C.

The fresh vegetable juice is one or more of mustard juice, cabbage juice, tomato juice and carrot juice.

The vegetables in step (3) are also pretreated as follows: the vegetables are washed with clean water, and the water on the surface is drained, cut into appropriate sizes and blanched with hot water.

The vegetable is one or more than two kinds of mustard greens, kohlrabi, cabbage, cabbage, potherb mustard, cucumber, beans, carrots, white radish, lettuce and pepper.

The fermented vegetables described in step (4) are put into polyethylene plastic bags or aluminum-plastic bags, and then heat-sealed after vacuumizing.

The fermented kimchi containing live mixed lactic acid bacteria prepared by the above method is vacuum-packed and stored at normal temperature for 6 months. The fermented kimchi contains Lactobacillus plantarum DMDL9010 and Lactobacillus caseisubsp.rhamnosus6013. The total number of live bacteria reaches 10 ⁵ -10 ⁶ cfu/g.

Compared with the prior art, the present invention has the following advantages and beneficial effects:

1. The present invention is a low-salt fermented pickle containing live mixed lactic acid bacteria, which is convenient, hygienic and nutritious.

2. The kimchi of the present invention contains live probiotics, the total number of live bacteria of Lactobacillusplantarum DMDL9010 and Lactobacilluscaseisubsp.rhamnosus6013 reaches 10 ⁵ -10 ⁶ cfu/g; the probiotics utilize the organic acids and antibacterial substances produced by the fermentation of nutrients in vegetables, which is beneficial to low-salt Preservation and preservation of fermented kimchi.

3. The present invention is clear with probiotics, its quality and concentration can be controlled artificially, the flavor of fermented kimchi is good, the production process is simple, industrialized production can be realized, the consistency of product quality in each batch is guaranteed, the production cycle is short, the product quality is stable, and it contains live probiotics, ready-to-eat.

4. The kimchi fermented by the technology of the present invention almost does not contain nitrite, which is far lower than the content of 10mg/Kg.

5. The salt content in fermented vegetable products is as low as 0.2% to 3.0%. This technology does not need to carry out complicated desalination work, simplifies the production process, and can realize mechanized operation.

6. Due to the fermentation of two kinds of lactic acid bacteria, the nitrite in vegetables is completely degraded, and the fermented kimchi will not undergo subsequent fermentation and produce gas swelling, which provides a stable quality of kimchi products during the shelf life. condition.

The Lactobacillus (Lactobacillus sp.) DMDL9010 has been preserved in the General Microbiology Center of the China Committee for Microbial Culture Collection on August 19, 2011, referred to as CGMCC, address: Datun Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences , the deposit number is CGMCCNo.5172. The strain has been disclosed in the patent publication number CN102978134A, and a preservation certificate has been submitted.

Description of drawings

Fig. 1 is the kimchi after adding two kinds of lactic acid bacteria to ferment in embodiment 1.

Fig. 2 is the kimchi after fermentation without adding lactic acid bacteria.

Detailed ways

In order to better understand the present invention, the present invention will be further described in detail below in conjunction with the examples, but the protection scope of the present invention is not limited to the range indicated by the examples.

Example 1

(1) Place the freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached 2.0×10 ⁸ CFU/mL; the Lactobacillus casei subsp. rhamnosus was purchased from China Industrial Microorganism Culture Collection and Management Center, numbered CICC6013, and named Lactobacillus caseisubsp.rhamnosus6013 (abbreviated as LCR6013).

(2) With a volume percentage of 5%, the seed liquids of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of viable bacteria reached 8.2×10 ⁸ CFU/mL and 7.9×10 ⁸ CFU/mL respectively; the formula of the working fermentation medium was: 85 parts of fresh mustard juice, 5 parts of tomato juice, 0.5 parts of vitamin C, NaCl1 .5 parts, distilled water to make up to 100 parts, sterilized and cooled for later use, which is the working fermentation medium.

(3) Select kohlrabi, cabbage, carrots and mustard greens that are fresh, moderately mature, free from borers, free from rot, and thick and firm.

(4) Remove the old, rotten and inedible parts, wash the sediment on the surface with clean water, drain the water on the surface, cut into appropriate sizes and blanch with hot water.

(5) According to the weight of 20 parts of vegetables, add a total of 10 parts of two kinds of lactic acid bacteria working fermentation liquid according to the ratio of 1:1, add 0.1 parts of vitamin C and 0.5 parts of NaCl, and dilute to 100 parts with sterile water , so that the vegetables are submerged in the liquid.

(6) Inject sterilized water around the sealing cover of the fermentation altar, seal the altar cover with water, isolate the oxygen in the air from entering, and ferment the temperature of the fermentation room at 37° C. for anaerobic fermentation for 48 hours; measure the nitrite content in the fermented kimchi, Refer to the method in the literature {Liu Dongmei, et al. Lactobacillus Caseisubsp.rhamnosus719 on the influence of nitrite in kimchi, Journal of South China University of Technology (Natural Science Edition), 2008, 36(7): 140-144}, no nitrite was detected Salt, the prepared kimchi is suitable for sourness and has the proper flavor and color of kimchi, as shown in Figure 1 and 2. Compared with the kimchi fermented without adding lactic acid bacteria, Lactobacillus DMDL9010 and Lactobacillus casei were added at a ratio of 1:1. The kimchi fermented by subspecies sugar has good color and luster, good flavor, no nitrite, and is a kind of safe kimchi.

(7) Put the cut vegetables into polyethylene plastic bags or aluminum-plastic bags, and heat seal them after vacuuming. Store the packaged fermented kimchi at room temperature for 6 months, take samples to constant volume, and use MRS medium to sample and analyze the number of viable lactic acid bacteria in it, which is 8.6×10 ⁶ cfu/g.

The effect of Lactobacillus DMDL9010 on degrading cholesterol in vivo:

(1) DMDL9010 was inoculated at 5% by volume in 200mL liquid MRS medium for activation, placed in a 37°C incubator for 24h, and again inoculated at 5% by volume in a fermenter for high-density anaerobic culture, After culturing for 18 hours at 37°C and pH 6.8, centrifuge at 8000 r/min and 4°C for 15 minutes, discard the supernatant, and collect the sludge. The volume ratio of trehalose (protectant) to the sludge is 1.5 :1 ratio, pre-freeze at -40°C for 5 hours to freeze evenly on the inner wall of the container, then carry out vacuum freeze-drying, after drying for 18-20 hours, rehydrate and measure the number of viable bacteria, DMDL9010 bacteria powder The number of viable bacteria was 9.30×10 ⁹ CFU/g.

(2) 50 8-week-old SPF grade SD (Sprague Dawley) rats, weighing 160-200 g, were randomly divided into 5 groups according to their body weight and blood lipid. All SD rats were raised by professionals, 5/cage. Under the conditions of a temperature of 22-24°C and a humidity of 50%-60%, the light is 12 hours and the darkness is 12 hours a day. During the experiment, the rats had free access to water and food, the normal group was fed with common feed, and the other groups were fed with high-fat feed (10% lard, 1% cholesterol, and 0.2% bile salt were added to the normal feed). Rats in each experimental group were intragastrically administered every morning. Both the normal group and the high-fat model group were intragastrically administered sterile water, and the positive group was intragastrically administered 1 mg/mL atorvastatin calcium tablet (also called Lipitor) aqueous solution. The dose group used 10 ⁹ CFU/ml DMDL9010 bacterial suspension for gavage, and the 9010 low-dose group used 10 ⁷ CFU/ml DMDL9010 bacterial suspension for gavage, and the gavage volume was 0.1mL/10gbw.d.

In the 8th week, all rats were fasted overnight, blood was collected by tail docking, centrifuged (3000r/min, 10min), and serum was separated. At week 10, all rats were fasted overnight, blood was collected from the abdominal cavity, and serum was obtained by centrifugation (3000 r/min, 10 min). Total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) contents were measured in serum with an automatic biochemical analyzer, and the results are shown in Table 1, Table 2.

As shown in Table 1, at the 8th week, it was found that although there was no significant difference in TC and TG among the groups, the TC and TG levels of the 9010 group were lower than those of the lipid model group, indicating that the 9010 group had no significant effect on hyperlipidemia. Symptoms have a certain palliative effect. Comparing LDL-C, it was found that although there was no significant difference among the groups, the LDL-C of all high-fat model groups was higher than that of the normal group, indicating that long-term feeding of high-fat diets led to low-density lipoprotein cholesterol levels in rats. increased the risk of cardiovascular disease in rats. However, the LDL-C of the experimental group fed with DMDL9010 bacterial solution was lower than that of the lipid model group, which indicated that DMDL9010 strain may have a certain preventive effect on cardiovascular diseases.

Table 1 Comparison of blood lipid test results of each group in the 8th week (mmol/L) (M±SD, n=10)

^※ There is a difference compared with the normal group (p<0.05) ^▲ There is a difference compared with the high-fat model group (p<0.05).

As shown in Table 2, at the 10th week, there was no significant difference in TC and LDL-C between the groups, but the TG level of the 9010 high-dose group was significantly lower than that of the high-fat model group, and its TC, LDL-C The level of LDL-C was between the normal group and the high-fat model group, indicating that the 9010 group could relieve hyperlipidemia and lower serum cholesterol levels in rats. Compared with HDL-C, it was found that the HDL-C of all high-fat model groups was lower than that of the normal group, and there was a significant difference (p<0.05) between the normal group. The reduction of high-density lipoprotein cholesterol increased the risk of arteriosclerosis in rats. Among them, the increase of HDL-C in the experimental group administered with DMDL9010 bacterial solution was more obvious, which indicated that DMDL9010 strain may have a certain effect on regulating arteriosclerosis.

Table 2 Comparison of blood lipid test results of each group in the 10th week (mmol/L) (M±SD, n=10)

^※ There is a difference compared with the normal group (p<0.05) ^▲ There is a difference compared with the high-fat model group (p<0.05).

(3) In the 10th week of the experiment, all rats were fasted overnight, and after the rats were killed, the liver was separated, and a certain amount of liver tissue was accurately weighed, and 10 mL of chloroform:methanol (2:1, V /V) Mixed solution, shake and mix, keep warm at 37°C for 30min, centrifuge (80000r/min, 10min, 4°C), carefully collect the chloroform layer liquid into a new EP tube, add 6mL normal saline, centrifuge (80000r/min, 10min, 4°C). After repeating the above steps again, collect the liquid in the bottom chloroform layer, dry it with a nitrogen blower, add 0.8 mL of isopropanol: Triton-100 (9:1, V/V) mixture to redissolve, vortex for 2 minutes, add 1.2mL of distilled water, and then vortexed for 2min to fully dissolve it, the resulting solution is the extract of total lipid in liver tissue, keep two aliquots for future use.

Accurately draw cholesterol standard solution (192.000mg/dL) and dilute to 1.920mg/mL, 1.536mg/mL, 0.768mg/mL, 0.384mg/mL, 0.192mg/mL, 0.000mg/mL respectively. Take 0.01 mL each and add it to 1 mL of working solution, mix well and incubate at 37°C for 6 min, measure the absorbance at 490 nm with a microplate reader, set up two replicate wells for each well, and draw a standard curve of cholesterol content. Sample determination was carried out according to the total cholesterol kit. Take 0.01mL of liver tissue total lipid extract and add it to 1mL working solution. After mixing, keep it warm at 37°C for 6min. Repeat hole.

Accurately draw triglyceride standard solution (200.0mg/dL) and dilute to 2.0mg/mL, 1.6mg/mL, 0.8mg/mL, 0.4mg/mL, 0.2mg/mL, 0.0mg/mL respectively. Take 0.01 mL each and add it to 1 mL of working solution, mix well and incubate at 37°C for 10 min, measure the absorbance at 490 nm with a microplate reader, set up two replicate wells for each well, and draw a standard curve for triglyceride content. The sample was measured according to the triglyceride kit. Take 0.01mL of the total lipid extract of liver tissue and add it to 1mL of the working solution. multiple holes.

The liver lipid detection results of the rats in each experimental group are shown in Table 3. The levels of TC and TG in the liver of the rats in each high-fat model group were higher than those in the normal group, with significant differences (p<0.05), indicating that excessive dietary sources Too much cholesterol will accumulate in the liver. TC and TG of the 9010 high-dose group decreased by 33.2% and 40.8% respectively in the high-fat model group in the 9010 group, and there was a significant difference (p<0.05), indicating that the DMDL9010 bacterial solution was High doses can effectively inhibit the accumulation of cholesterol in the liver and reduce the content of triglycerides in the liver.

Table 3 Detection results of liver lipids in rats in each experimental group (M±SD, n=10)

Example 2

(1) The freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached 5.0×10 ⁸ CFU/mL;

(2) With a volume percentage of 10%, the seed liquids of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of viable bacteria reached 2.5×10 ⁸ CFU/mL and 5.5×10 ⁸ CFU/mL respectively; the formulation of the working fermentation medium was: 85 parts of fresh cabbage juice, 5 parts of tomato juice, 3.0 parts of vitamin C, NaCl1 .5 parts, distilled water to make up to 100 parts, sterilized and cooled for later use, which is the working fermentation medium.

(3) Choose cabbage, cucumber, lettuce that are fresh, moderately ripe, free from borers, free from rot, and thick and firm;

(4) Remove the old, rotten and inedible parts, and wash the silt on the surface with clean water, drain the water on the surface, cut into appropriate sizes and blanch with hot water;

(5) According to the weight of 30 parts of vegetables, add 5 parts of two kinds of lactic acid bacteria working fermentation broth according to the ratio of 3:1, add 1.5 parts of vitamin C and 3.0 parts of NaCl, and dilute to 100 parts with sterile water , try to keep the vegetables submerged in the liquid;

(6) Inject sterilized water around the sealing cover of the fermentation altar, seal the altar cover with water, isolate the oxygen in the air from entering, and control the temperature of the fermentation room at 30°C for anaerobic fermentation for 24 hours; measure the nitrite content in pickles after fermentation , no nitrite was detected, the prepared kimchi was suitable for sourness, and had the proper flavor and color of kimchi.

(7) Put the cut vegetables into polyethylene plastic bags or aluminum-plastic bags, and heat seal them after vacuuming. Store the packaged fermented kimchi at room temperature for 6 months, take samples to constant volume, and use MRS medium to sample and analyze the number of viable lactic acid bacteria in it, which is 6.7×10 ⁶ cfu/g.

Example 3

(1) The freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached 4.5×10 ⁸ CFU/mL;

(2) With a volume percentage of 1%, the seed liquids of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of viable bacteria reached 5.0×10 ⁸ CFU/mL and 4.2×10 ⁸ CFU/mL respectively; the formula of the working fermentation medium was: 70 parts of fresh mustard juice, 5 parts of tomato juice, 5 parts of carrot juice, vitamin C1 .5 parts, 3.0 parts of NaCl, distilled water to 100 parts, cooled after sterilization, standby, is the working fermentation medium.

(3) Select potherb mustard, peppers and mustard greens that are fresh, moderately mature, free from borers, free from rot, and have thick and firm flesh;

(4) Remove the old, rotten and inedible parts, and wash the silt on the surface with clean water, drain the water on the surface, cut into appropriate sizes and blanch with hot water;

(5) According to the weight of 40 parts of vegetables, add 5 parts of two kinds of lactic acid bacteria working fermentation broth according to the ratio of 1:1, add 1.0 parts of vitamin C and 3.5 parts of NaCl, and dilute to 100 parts with sterile water , try to keep the vegetables submerged in the liquid;

(6) Inject sterilized water around the sealing cover of the fermentation altar, seal the altar cover with water, isolate the oxygen in the air from entering, and control the temperature of the fermentation room at 30° C. for anaerobic fermentation for 72 hours; measure the nitrite content in the fermented pickles, No nitrite was detected, and the prepared kimchi was suitable for sourness and had the proper flavor and color of kimchi.

(7) Put the cut vegetables into polyethylene plastic bags or aluminum-plastic bags, and heat seal them after vacuuming. Store the packaged fermented kimchi at room temperature for 6 months, take samples to constant volume, and analyze the number of live lactic acid bacteria in MRS medium, which is 8.6×10 ⁶ cfu/g.

Example 4

(1) The freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached 2.7×10 ⁸ CFU/mL;

(2) With a volume percentage of 5%, the seed liquids of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of viable bacteria reaches 3.3×10 ⁸ CFU/mL and 2.9×10 ⁸ CFU/mL respectively; the formula of the working fermentation medium is: 90 parts of fresh mustard juice, 1.5 parts of vitamin C, 1.5 parts of NaCl, distilled water Dilute to 100 parts, sterilize and cool down for later use, which is the working fermentation medium.

(3) Select mustard greens that are fresh, moderately mature, free from moths, rotten, and thick and firm;

(4) Remove the old, rotten and inedible parts, and wash the silt on the surface with clean water, drain the water on the surface, cut into appropriate sizes and blanch with hot water;

(5) According to the weight of 50 parts of vegetables, add 1 part of two kinds of lactic acid bacteria working fermentation liquid according to the ratio of 2:1, add 0.5 parts of vitamin C and 1.5 parts of NaCl, and dilute to 100 parts with sterile water , try to keep the vegetables submerged in the liquid;

(6) Inject sterilized water around the sealing cover of the fermenting altar, seal the altar cover with water, isolate the oxygen in the air from entering, and control the temperature of the fermenting chamber at 30° C. for anaerobic fermentation for 60 h; measure the nitrite content in the fermented kimchi, No nitrite was detected, and the prepared kimchi was suitable for sourness and had the proper flavor and color of kimchi.

(7) Put the cut vegetables into polyethylene plastic bags or aluminum-plastic bags, and heat seal them after vacuuming. Store the packaged fermented kimchi at room temperature for 6 months, take samples to constant volume, and use MRS medium to sample and analyze the number of viable lactic acid bacteria in it, which is 9.0×10 ⁵ cfu/g.

Example 5

(1) Place the freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached 2.0×10 ⁸ CFU/mL;

(2) With the volume percentage of 7%, the seed liquid of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of viable bacteria reached 4.7×10 ⁸ CFU/mL and 7.9×10 ⁸ CFU/mL respectively; the formula of the working fermentation medium was: 50 parts of fresh mustard juice, 25 parts of cabbage juice, 10 parts of tomato juice, carrot 5 parts of juice, 2.5 parts of vitamin C, 2.0 parts of NaCl, distilled water to 100 parts, sterilized and cooled for later use, which is the working fermentation medium.

(3) Select kohlrabi and mustard greens that are fresh, moderately mature, free from moths, rotten, and thick and firm;

(4) Remove the old, rotten and inedible parts, and wash the silt on the surface with clean water, drain the water on the surface, cut into appropriate sizes and blanch with hot water;

(5) According to the weight of 45 parts of vegetables, add 5 parts of two kinds of lactic acid bacteria working fermentation broth according to the ratio of 2:1, add 2.0 parts of vitamin C and 0.2 parts of NaCl, and dilute to 100 parts with sterile water , try to keep the vegetables submerged in the liquid;

(6) inject sterilized water around the sealing cover of the fermenting altar, seal the altar cover with water, isolate the oxygen in the air from entering, and ferment the temperature of the fermenting chamber at 25° C. for anaerobic fermentation for 72 hours; measure the nitrite content in the fermented kimchi, No nitrite was detected, and the prepared kimchi was suitable for sourness and had the proper flavor and color of kimchi.

(7) Put the cut vegetables into polyethylene plastic bags or aluminum-plastic bags, and heat seal them after vacuuming. Store the packaged fermented kimchi at room temperature for 6 months, take a sample to make up the volume, and use the sample to analyze the number of viable lactic acid bacteria in the MRS medium, which is 2.5×10 ⁶ cfu/g.

Example 6

(1) The freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached 2.0×10 ⁸ CFU/mL;

(2) With the volume percentage of 8%, the seed liquids of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of viable bacteria reached 2.3×10 ⁸ CFU/mL and 6.2×10 ⁸ CFU/mL respectively; the formula of the working fermentation medium was: 75 parts of fresh cabbage juice, 5 parts of tomato juice, 5.0 parts of carrot juice, vitamin C0 .5 parts, 1.5 parts of NaCl, distilled water to 100 parts, cooled after sterilization, standby, that is, the working fermentation medium.

(3) Select mustard greens, white radishes, and beans that are fresh, moderately mature, free from borers, free from rot, and have thick and firm flesh;

(4) Remove the old, rotten and inedible parts, and wash the silt on the surface with clean water, drain the water on the surface, cut into appropriate sizes and blanch with hot water;

(5) According to the weight of 30 parts of vegetables, add a total of 8 parts of two kinds of lactic acid bacteria working fermentation broth according to the ratio of 5:1, add 2.0 parts of vitamin C and 5.0 parts of NaCl, and dilute to 100 parts with sterile water , try to keep the vegetables submerged in the liquid;

(6) inject sterilized water around the sealing cover of the fermenting altar, seal the altar cover with water, isolate the oxygen in the air from entering, and ferment the temperature of the fermenting chamber at 20° C. for anaerobic fermentation for 68 hours; measure the nitrite content in the fermented kimchi, No nitrite was detected, and the prepared kimchi was suitable for sourness and had the proper flavor and color of kimchi.

(7) Put the cut vegetables into polyethylene plastic bags or aluminum-plastic bags, and heat seal them after vacuuming. Store the packaged fermented kimchi at room temperature for 6 months, take samples to constant volume, and analyze the number of viable lactic acid bacteria in the MRS medium, which is 3.7×10 ⁵ cfu/g.

Claims (7)

1. a kind of preparation method that contains the kimchi of mixing lactic acid bacteria, it is characterized in that, wherein mixing lactic acid bacteria comprises lactobacillus (Lactobacillus sp.) DMDL9010 and Lactobacillus caseis subsp. ) DMDL9010 was deposited on August 19, 2011 in the General Microorganism Center of China Committee for Culture Collection of Microorganisms, referred to as CGMCC, and the preservation number is CGMCC No.5172. 2. The method according to claim 1, characterized in that, comprising the steps of: (1) Place the freeze-dried powders of Lactobacillus DMDL9010 and Lactobacillus casei subsp. The number of bacteria reached above 10 ⁸ CFU/mL; (2) Place the seed liquids of Lactobacillus DMDL9010 and Lactobacillus casei subspecies rhamnose in the fermentation medium at a volume percentage of 1% to 10%, and culture them statically at 30°C to 40°C for 18h to 36h, Prepare a fermentation broth so that the number of viable bacteria reaches 10 ⁸ CFU/mL or more; the formulation of the fermentation medium is: 80-90 parts of fresh vegetable juice, 0.5-3.0 parts of vitamin C, 0.5-3.0 parts of NaCl Parts, dilute to 100 parts with sterile water, after sterilization and cooling, it is the fermentation medium; (3) In parts by weight, 20 to 40 parts of vegetables are inserted into 3 to 10 parts of the fermentation broth of Lactobacillus DMDL9010 and Lactobacillus casei subsp. Add 0.1 to 2.0 parts of vitamin C and 0.2 to 5.0 parts of NaCl each, dilute to 100 parts with sterile water, and immerse the vegetables in the liquid; (4) Carry out anaerobic fermentation under sealed conditions, and ferment for 24-72 hours at a temperature of 20-40°C. 3. The method according to claim 2, characterized in that, said fresh vegetable juice is one or more of mustard juice, cabbage juice, tomato juice and carrot juice. 4. The method according to claim 2 or 3, characterized in that the vegetables in step (3) are also pretreated as follows: the vegetables are washed with clear water, and the moisture on the surface is drained, cut into appropriate sizes and used Blanch in hot water. 5. The method according to claim 4, wherein the vegetables are one or more of mustard greens, kohlrabi, cabbage, cabbage, potherb mustard, cucumber, beans, carrots, white radish, lettuce and pepper . 6. The method according to claim 5, characterized in that the fermented vegetables in step (4) are packed into polyethylene plastic bags or aluminum-plastic bags, and then heat-sealed after vacuumizing. 7. The kimchi containing mixed lactic acid bacteria prepared by the method according to any one of claims 1 to 6.