Lactobacillus plantarum and application thereof in fermented sour cucumber

Abstract

The application provides a lactobacillus plantarum and application thereof in fermented sour cucumbers, and belongs to the technical field of microbial fermentation. The lactobacillus plantarum is preserved in the Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC No: 66118. The application also provides a method for preparing the sour cucumbers by fermentation of the lactobacillus plantarum, which comprises the following steps: 1) pickling and dehydrating cucumbers to obtain dehydrated cucumbers; and 2) mixing the dehydrated cucumbers with salt brine and fermenting to obtain the sour cucumbers. The lactobacillus plantarum has excellent fermentation characteristics, and when the lactobacillus plantarum is applied to cucumber fermentation, the fermentation rate can be improved, the fermentation period can be shortened, the crispness of the cucumbers can be enhanced, and the product quality can be stabilized.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of microbial fermentation, and particularly relates to a plant lactobacillus and application thereof in fermented sour cucumbers. BACKGROUND

[0002] Cucumbers are widely planted in many places due to their high yield, low price and easy storage. Fermented cucumbers have a long history as a traditional fermented vegetable product. Fermented sour cucumbers are crisp and tender in texture, juicy and sweet in taste, aromatic and delicious, and contain rich ingredients such as protein, fat, sugar, various vitamins, cellulose, and calcium, phosphorus, iron, potassium, sodium, magnesium, etc. In particular, the fine cellulose contained in cucumbers can reduce the content of cholesterol and triglycerides in the blood, promote intestinal peristalsis, accelerate waste excretion, and improve human metabolism, thereby being favored by more and more consumers.

[0003] At present, the production process of sour cucumbers mostly adopts the traditional processing method, i.e. natural fermentation. This method not only has a complicated process and a long production cycle, but also faces problems such as short shelf life, softening of texture and unstable product quality due to the influence of temperature and environmental factors during fermentation. Therefore, there is an urgent need for sour cucumber deep processing technologies and products with high technical content, good protection of nutritional functional ingredients and high added value. After lactic acid bacteria fermentation, cucumbers not only have improved flavor, but also can regulate intestinal flora and promote the absorption of nutrients by the intestinal tract, and have the effects of enhancing immunity, reducing blood sugar, reducing blood pressure, resisting tumors and preventing cancer. Sour cucumbers with lactic acid bacteria as a ferment integrate the nutritional value of cucumbers and the health care function of lactic acid bacteria. Lactic acid bacteria fermentation has been widely used in fermented vegetables such as radish, cowpea and sauerkraut, but there are few studies on inoculation of lactic acid bacteria into cucumber fermentation system at home and abroad. Therefore, strain selection is one of the key factors for improving the quality of lactic acid bacteria fermented cucumbers. At present, there are few special strains of lactic acid bacteria used for fermentation of sour cucumbers in research, and the strains used have poor adaptability to cucumber fermentation, resulting in problems such as low number of live bacteria, soft texture and the like in existing lactic acid bacteria fermented cucumber products. Therefore, screening of special strains of lactic acid bacteria with excellent fermentation performance for sour cucumbers is the primary task of lactic acid bacteria fermentation technology for sour cucumbers. SUMMARY

[0004] In view of this, the purpose of the present application is to solve the problems of long fermentation cycle, short shelf life, softening of texture and unstable product quality in natural fermentation of sour cucumbers. A plant lactobacillus with excellent fermentation characteristics is screened, which is applied to the preparation of sour cucumbers, can improve the fermentation rate, shorten the fermentation cycle, enhance the crispness of cucumbers and stabilize the product quality of sour cucumbers.

[0005] The application provides a Lactiplantibacillus plantarum SAAS-B-MRS-20201016-2 which is preserved in the Guangdong Microbial Culture Collection Center and has a preservation number of GDMCC No: 66118.

[0006] The application also provides application of the Lactiplantibacillus plantarum SAAS-B-MRS-20201016-2 in fermenting preparation of sour cucumbers.

[0007] Preferably, the raw cucumbers of the sour cucumbers are Sichuan green small jade.

[0008] The application provides a method for fermenting preparation of sour cucumbers by using the Lactiplantibacillus plantarum SAAS-B-MRS-20201016-2, and the method comprises the following steps:

[0009] 1) pickling and dehydrating cucumbers to obtain dehydrated cucumbers;

[0010] 2) mixing the dehydrated cucumbers with salt brine and fermenting to obtain sour cucumbers;

[0011] The salt brine comprises 3%-10% (v / v) of Lactiplantibacillus plantarum SAAS-B-MRS-20201016-2 bacterial liquid; the viable bacterial concentration of the Lactiplantibacillus plantarum SAAS-B-MRS-20201016-2 bacterial liquid is 10 6 ~ 10 7 CFU / mL.

[0012] Preferably, the cucumbers are mixed with salt and then pickled in step 1), the pickling temperature is 10-35 DEG C, and the pickling time is 2-4 h.

[0013] Preferably, the dehydration amount of the dehydration in step 1) is controlled to be 20%-25%.

[0014] Preferably, the ratio of the dehydrated cucumbers to the salt brine in step 2) is 1 kg:(0.5-1.5) L.

[0015] Preferably, the fermentation temperature in step 2) is 18-22 DEG C, and the fermentation time is 5-10 days.

[0016] Preferably, the fermentation is ended when the total acid content reaches 0.6%-1.5% in step 2).

[0017] The application provides the sour cucumbers obtained by the method.

[0018] Compared with the prior art, the application has the following beneficial effects: a plant lactobacillus SAAS-B-MRS-20201016-2 with excellent fermentation characteristics is screened from pickled water, and the application of the plant lactobacillus to cucumber fermentation can improve the fermentation rate, shorten the fermentation period, enhance the crispness of the cucumber and stabilize the product quality.

[0019] The plant lactobacillus SAAS-B-MRS-20201016-2 provided by the application has strong acid tolerance, and the number of lactic acid bacteria, total acid, organic acid, hardness and moisture content of fermented cucumber are studied by taking the pickled vegetable fermentation bacteria as a control. 9 The results show that the plant lactobacillus can grow well in the fermented cucumber, and the number of viable bacteria reaches 6.4x10 8 CFU / g, which is better than 1.2x10 8 CFU / g of the control pickled vegetable fermentation bacteria, and the difference is significant (P<0.05); the total acid of the acid cucumber containing the plant lactobacillus is 1.24%, while the total acid of the acid cucumber containing the control pickled vegetable fermentation bacteria is 1.15%; the lactic acid of the acid cucumber containing the plant lactobacillus is 7.92g / kg, and the acetic acid is 2.70g / kg, which are higher than 6.99g / kg of the lactic acid and 1.77g / kg of the acetic acid of the control acid cucumber; the hardness of the acid cucumber containing the plant lactobacillus is 452.183g, which is higher than 421.287g of the control acid cucumber; and the moisture content of the acid cucumber containing the plant lactobacillus is 95.05%, which is lower than 95.53% of the control acid cucumber. BRIEF DESCRIPTION OF DRAWINGS

[0020] Fig. 1 The number of lactic acid bacteria in the cucumber fermentation process of example 1 is shown in the table.

[0021] Fig. 2 The pH value in the cucumber fermentation process of example 1 is shown in the table.

[0022] Fig. 3 The hardness in the cucumber fermentation process of example 1 is shown in the table.

[0023] Fig. 4 The colony morphology of the plant lactobacillus obtained by separation in example 1 is shown in the figure.

[0024] Fig. 5 The microscopic examination of the plant lactobacillus obtained by separation in example 1 is shown in the figure.

[0025] Fig. 6 The acid tolerance of the plant lactobacillus obtained by separation in example 1 is shown in the table.

[0026] BIOLOGICAL PRESERVATION

[0027] The plant lactiplantibacillus plantarum provided by the present application is preserved in the Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC No: 66118, the preservation date is April 10, 2025, and the preservation address is No. 59, Building 5, Guangzhou Xianlie Middle Road, Guangzhou, China. DETAILED DESCRIPTION

[0028] The present application provides a plant lactiplantibacillus plantarum, which is preserved in the Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC No: 66118. In the present application, the plant lactiplantibacillus plantarum colony is milky white, the colony surface is smooth, and the edge is neat; the gram staining result shows that it is a gram-positive bacterium, the cell is rod-shaped, and the bacterial body is single, paired or chain-shaped. The 16S rDNA sequence of the plant lactiplantibacillus plantarum provided by the present application is shown in SEQ ID No. 1; the plant lactiplantibacillus plantarum provided by the present application has certain antioxidant capacity and strong acid tolerance.

[0029] The present application also provides the application of the plant lactiplantibacillus plantarum in the fermentation preparation of sour cucumbers. In the present application, the raw material cucumber of the sour cucumber is preferably a Sichuan green small jade.

[0030] The present application provides a method for preparing a sour cucumber by fermentation of the plant lactiplantibacillus plantarum, comprising the following steps: 1) pickling and dehydrating cucumbers to obtain dehydrated cucumbers; 2) mixing and fermenting the dehydrated cucumbers with salt brine to obtain sour cucumbers.

[0031] In the present application, the cucumbers are pickled to obtain dehydrated cucumbers. In the present application, the cucumbers are preferably pretreated, and the pretreatment includes removing inedible parts, washing, draining and cutting. The present application does not have special limitations on the specific operation of the pretreatment, and the conventional operation in the art can be used. After the pretreatment of the cucumbers, the present application is pickled; the cucumbers are mixed with salt for pickling, and the amount of salt is 3% to 5% of the mass of the cucumbers, preferably 4%; the temperature of the pickling is preferably 10 to 35℃, and the time of the pickling is preferably 2 to 4h. The present application realizes dehydration through the pickling, and the dehydration amount of the dehydration is preferably controlled at 20% to 25%, and the present application obtains dehydrated cucumbers after the dehydration.

[0032] The acid cucumber is obtained by mixing the dehydrated cucumber with salt brine and fermentation. In the present application, the mixing ratio of the dehydrated cucumber and the salt brine is 1 kg: (0.5-1.5) L, preferably 1 kg: (0.8-1.2) L, and more preferably 1 kg: 1 L. In the present application, the salt brine contains 3%-10% (v / v) of Lactobacillus plantarum bacterial solution, preferably 4%-7%, and more preferably 5%-6%; the viable bacterial concentration of the Lactobacillus plantarum bacterial solution is 10 6 ~10 7 CFU / mL. In the present application, the salt brine preferably further contains the following components in the following mass percentages: 1.5%-2.5% of salt, 0.3%-0.7% of pepper, 0.08%-0.12% of garlic, and 0.08%-0.12% of Sichuan pepper. In the present application, the above raw materials are preferably added to cold water to be fully dissolved to obtain the salt brine.

[0033] In the present application, the fermentation temperature is preferably 18-22℃, further preferably 19-31℃, and more preferably 20℃; the fermentation time is preferably 5-10 days, further preferably 6-9 days, and more preferably 7 days. In the present application, the fermentation is preferably ended when the total acid content is 0.6%-1.5%.

[0034] The present application also provides the acid cucumber prepared by the method.

[0035] The technical solutions provided by the present application will be described in detail below in conjunction with the examples, but they should not be understood as limiting the scope of protection of the present application.

[0036] Example 1

[0037] Materials

[0038] Secondary fermentation water of pickled Chinese cabbage: collected by a food company in Sichuan;

[0039] Conventional fermentation agent: Shangchuan pickled vegetable fermentation bacteria, purchased from Shangchuan Biotechnology Co., Ltd. in Shunde, Guangdong.

[0040] MRS agar medium: 10.0 g / L of proteose peptone, 5.0 g / L of beef extract powder, 4.0 g / L of yeast extract powder, 20.0 g / L of glucose, 2.0 g / L of potassium phosphate dibasic, 2.0 g / L of trisodium citrate, 5.0 g / L of sodium acetate, 0.2 g / L of magnesium sulfate, 0.05 g / L of manganese sulfate, 15.0 g / L of agar, and 1.0 g / L of Tween 80; the pH value is 7.0±0.2.

[0041] MRS meat culture medium: peptone 10.0 g / L, beef extract powder 5.0 g / L, yeast extract powder 4.0 g / L, glucose 20.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, ammonium citrate tribasic 2.0 g / L, sodium acetate 5.0 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, Tween 80 1.0 g / L; pH 7.0 ± 0.2.

[0042] Screening of lactic acid bacteria with excellent fermentation characteristics

[0043] (1) Initial screening of lactic acid bacteria with excellent fermentation characteristics

[0044] Weigh 25g of pickled mustard greens, add 225mL of sterile saline, and mix well using a homogenizer. -1 Dilute the liquid by adding 1 mL of the above dilution to 9 mL of sterile saline and mix well to obtain 10 -2 The dilution solution was diluted in order to obtain 10 -3 , 10 -4 , 10 -5 Dilution: Take 100 μL of the above dilution and spread it on an MRS agar plate containing 2% CaCO3. Incubate at 36°C for 72 hours. Pick a single colony with a large calcium-soluble circle and streak it on an MRS agar plate for purification 2 to 3 times.

[0045] (2) Rescreening of lactic acid bacteria with excellent fermentation characteristics

[0046] Wash cucumbers, drain surface moisture, and cut into 1-2 cm round pieces for later use. Separate and purified single bacteria are inoculated into cucumber material at a 1% inoculum rate; the cucumber material comprises cucumbers, salt, and water; the salt content is 6% of the cucumber mass, and the mass of water is equal to the cucumber mass. The cucumber material and bacteria are placed in a 3-jin earthenware jar and fermented at 20°C for 20 days. The pH of the fermentation broth, the number of lactic acid bacteria, and the firmness of the cucumbers are measured on days 3, 6, 10, and 20 of fermentation.

[0047] The pH was measured using a pH meter; the lactic acid bacteria count was measured using an MRS plate count; and the hardness was measured using a texture analyzer with a p2 probe, a measuring speed of 1.5 mm / sec, and a test mode compression of 50%.

[0048] The results are as follows Figs. 1-3 As shown, the lactic acid bacteria content gradually increased during the fermentation process and reached a stable state after 3 days. The acidity trend during fermentation was consistent with that of the lactic acid bacteria, which may be due to the acid production of the lactic acid bacteria, which decreased the pH. The firmness of the cucumber increased slightly during the fermentation process, but the difference was not significant. These results indicate that the lactic acid bacteria-fermented cucumbers have a high lactic acid bacteria content and rapid acid production, making them suitable for fermented cucumber production and enhancing the firmness of pickled cucumbers.

[0049] Identification of lactic acid bacteria

[0050] (1) Morphological observation. The colonies on the MRS agar medium were picked. Smear, gram staining, and observation of bacterial shape, size, and gram staining reaction under a microscope; observation of spores, morphology, and attachment position.

[0051] After the lactic acid bacteria were cultured on the MRS plate for 48 h, the colony morphology and cell morphology were observed, and the results are shown in Fig. 2 The results show that the colony is milky white, the colony surface is smooth, and the edge is neat; the gram staining reaction observed under a microscope is shown in Fig. 3 , gram-positive bacteria, and the cells are rod-shaped, and the bacterial bodies are arranged in single, paired, and chain forms.

[0052] (2) 16S r DNA identification. The primers are 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R: 5'-GGTTACCTTGTTACGACTT-3'; the PCR reaction system is 25 μL, including 12.5 μL of Mix, 0.5 μL of 27F, 0.5 μL of 1492R, and 11.5 μL of ddH2O; the PCR conditions are pre-denaturation of DNA double strands at 94 ℃ for 10 min; denaturation at 94 ℃ for 30 s; annealing at 50 ℃ for 30 s; extension at 72 ℃ for 80 s, and cycling for 29 times, and finally maintaining at 72 ℃ for 7 min.

[0053] Agar gel electrophoresis of the amplification product:

[0054] 0.5 g of agarose was dissolved in 50 mL of TBE buffer, and 5 μL of EB dye was added after heating and dissolving; after the gel was injected and solidified, 5 μL was added to each sample well; it was run at a voltage of 120 V for 20 min; the gel plate was observed under UV to observe the image, and the PCR product with clear bands was selected for sequencing by Beijing Qianke Biological Technology Co., Ltd.; 16S r DNA sequence analysis and identification were performed, the sequencing results were submitted to Genbank for blast comparison analysis, and the results showed that the homology with Lactiplantibacillus plantarum reached 100%. Combined with the morphological characteristics of the bacteria, the strain was identified as Lactiplantibacillus plantarum, and the 16S r RNA of the bacteria is as follows:

[0055]

[0056] Characteristics of Lactobacillus plantarum

[0057] (1) Antioxidant capacity of Lactobacillus plantarum

[0058] The purified Lactobacillus plantarum was inoculated into 100 mL of MRS broth at an inoculation amount of 1%, and then incubated at 36°C for 24 h. The supernatant was obtained by centrifugation at 12,000 r / min for 20 min.

[0059] DPPH scavenging capacity: 2 mL of the supernatant was added with 2 mL of DPPH solution at a concentration of 0.1 mmol / L. The mixture was reacted at room temperature for 30 min in the dark, centrifuged at 6,000 r / min for 10 min, and the absorbance of the supernatant was measured at 517 nm. Meanwhile, a control group and a blank group were set. The DPPH scavenging rate of the Lactobacillus plantarum was 1578.779 μg Trolox / mL.

[0060] Hydroxyl radical scavenging capacity: 1 mL of the supernatant was added with 1 mL of ferrous sulfate solution at a concentration of 9 mmol / L, 1 mL of salicylic acid ethanol solution at a concentration of 9 mmol / L, and 1 mL of hydrogen peroxide solution at a concentration of 9 mmol / L. The mixture was reacted at room temperature for 30 min in the dark, centrifuged at 6,000 r / min for 10 min, and the absorbance of the supernatant was measured at 510 nm. Meanwhile, a control group and a blank group were set. The hydroxyl radical scavenging rate of the Lactobacillus plantarum was 51.9 U / mL.

[0061] (2) Acid tolerance capacity

[0062] The activated Lactobacillus plantarum was inoculated into MRS liquid medium containing acetic acid and citric acid at concentrations of 0%, 0.4%, 0.6%, 0.8%, and 1.2% at an inoculation amount of 1%, and then incubated at 37°C for 24 h. The number of Lactobacillus plantarum was counted, and the results are shown in Table 6. When the concentrations of acetic acid and citric acid were lower than 0.4%, they had no effect on the growth of Lactobacillus plantarum. When the concentrations were higher than 0.4%, the growth of Lactobacillus plantarum was inhibited to a certain extent, and the inhibitory effect of citric acid was greater than that of acetic acid. The results showed that the Lactobacillus plantarum had strong acid tolerance, especially to acetic acid.

[0063] Example 2

[0064] Fermented sour cucumber containing Lactobacillus plantarum

[0065] (1) Pretreatment of Sichuan green small cucumber: The cucumber was trimmed to remove the inedible parts at both ends, washed, and then drained. The cucumber was cut in half, and then drained again to obtain the raw material.

[0066] (2) Cucumber pickling and dehydration: Mix the pretreated cucumber raw materials with 4% salt and stir evenly, then place it in a temperature of 10-35°C for 3h of pickling, and then dehydrate the pickled cucumber, with the dehydration amount controlled at 20-25%.

[0067] (3) Salt brine preparation: Take 5% (v / v) plant lactobacillus, 2% (m / v) salt, 0.5% (m / v) pepper, 0.1% (m / v) garlic, and 0.1% (m / v) Sichuan pepper, and add cold water to dissolve them thoroughly, and you get salt brine.

[0068] (4) Fermentation culture

[0069] Mix the pretreated cucumber from step (2) with the salt brine from step (3) at a ratio of 1:1 (m / v), and place it in a 20°C fermentation environment for 7 days, until the total acid reaches 0.6-1.5%, and you get fermented sour cucumber containing the bacteria.

[0070] Comparative Example 1

[0071] (1) Pretreatment of Sichuan green small cucumber: Remove the inedible parts from the cucumbers, wash and drain the water, then cut the cucumbers in half and drain the water, and you get the raw materials, ready for use.

[0072] (2) Cucumber pickling and dehydration: Mix the pretreated cucumber raw materials with 4% salt and stir evenly, then place it in a temperature of 10-35°C for 3h of pickling, and then dehydrate the pickled cucumber, with the dehydration amount controlled at 20-25%.

[0073] (3) Salt brine preparation: Take 5% (v / v) conventional fermenting agent, 2% (m / v) salt, 0.5% (m / v) pepper, 0.1% (m / v) garlic, and 0.1% (m / v) Sichuan pepper, and add cold water to dissolve them thoroughly, and you get salt brine.

[0074] (4) Fermentation culture

[0075] Mix the pretreated cucumber from step (2) with the salt brine from step (3) at a ratio of 1:1 (m / v), and place it in a 20°C fermentation environment for 7 days, until the total acid reaches 0.6-1.5%, and you get fermented sour cucumber.

[0076] Compare the quality of the fermented sour cucumber containing plant lactobacillus in Example 2 with the fermented sour cucumber containing conventional fermenting agent in Comparative Example 1.

[0077] Determine the number of lactic acid bacteria, total acid, organic acid, hardness, and moisture content of the two groups of sour cucumber.

[0078] The number of lactic acid bacteria is determined by the method in GB 4789.35-2023.

[0079] The total acid was determined by the potentiometric titration method in the national standard GB12456-2021.

[0080] The organic acid was determined by high performance liquid chromatography, and the chromatographic column was: HPX-87H, 300 mm x 7.8 mm; sample size: 5 μL; mobile phase: 0.004 M sulfuric acid; flow rate: 0.6 mL / min; detector: UV@215 nm

[0081] The hardness was determined by a texture analyzer, probe P2, measurement speed 1.5 mm / sec, and strain 50%.

[0082] The moisture content was determined by the direct drying method in the national standard GB5009.3-2016.

[0083] The results show that the plant lactobacillus can grow well in the fermented cucumber, and the viable cell count reaches 6.4 x 10 9 CFU / g, which is better than 1.2 x 10 8 CFU / g of the control pickle fermentation bacteria, and the difference is significant (P<0.05); the total acid of the sour cucumber containing the plant lactobacillus is 1.24%, while the total acid of the sour cucumber containing the control pickle fermentation bacteria is 1.15%; the lactic acid of the sour cucumber containing the plant lactobacillus is 7.92 g / kg, and the acetic acid is 2.70 g / kg, which are higher than 6.99 g / kg of the lactic acid and 1.77 g / kg of the acetic acid of the control sour cucumber; the hardness of the sour cucumber containing the plant lactobacillus is 452.183 g, which is higher than 421.287 g of the control sour cucumber; the moisture content of the sour cucumber containing the plant lactobacillus is 95.05%, which is lower than 95.53% of the control sour cucumber, indicating that the moisture content of the sour cucumber containing the plant lactobacillus is lower. It is proved that the plant lactobacillus has a wide application prospect in the fermented sour cucumber with high acid production, low moisture content and high cucumber crispness.

[0084] The above only describes the preferred embodiments of the present application, and it should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A Lactobacillus plantarum SAAS-B-MRS-20201016-2, characterized in that It was deposited in Guangdong Provincial Microbiological Culture Collection with the deposit number GDMCC No: 66118.

2. Use of the plant lactobacillus SAAS-B-MRS-20201016-2 described in claim 1 in fermenting and preparing pickled cucumbers.

3. The use according to claim 2, characterized in that The raw material cucumber of the pickled cucumber is Sichuan green cucumber.

4. A method for preparing pickled cucumbers by fermenting Lactobacillus plantarum SAAS-B-MRS-20201016-2 according to claim 1, characterized in that: The following steps are involved: 1) pickling and dehydrating the cucumbers to obtain dehydrated cucumbers; 2) mixing the dehydrated cucumbers with brine and fermenting them to obtain pickled cucumbers; The brine contains 3% to 10% (v / v) of Lactobacillus plantarum SAAS-B-MRS-20201016-2 bacterial solution; the live bacteria concentration of the Lactobacillus plantarum SAAS-B-MRS-20201016-2 bacterial solution is 10 6 ~10 7 CFU / mL.

5. The method according to claim 4, characterized in that Step 1) The cucumbers are mixed with salt and then pickled at a temperature of 10 to 35° C. for 2 to 4 hours.

6. The method according to claim 4 or 5, characterized in that The dehydration amount of the dehydration in step 1) is controlled at 20% to 25%.

7. The method according to claim 4, characterized in that Step 2) The dehydrated cucumbers and the brine are mixed in a ratio of 1 kg: (0.5-1.5) L.

8. The method according to claim 4 or 7, characterized in that Step 2) The fermentation temperature is 18-22° C. and the fermentation time is 5-10 days.

9. The method according to claim 8, characterized in that Step 2) When the total acid content of the fermentation product is 0.6% to 1.5%, the fermentation is terminated.

10. Pickled cucumbers prepared by the method according to any one of claims 4 to 9.

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