Application of lactobacillus plantarum SAAS-B-PCA-202043-6 in inhibition of aerogenesis saccharomycetes and application of lactobacillus plantarum SAAS-B-PCA-202043-6 in preparation of bagged fermented pepper
By inoculating bagged fermented chili peppers with Lactobacillus plantarum SAAS-B-PCA-202043-6, the problem of bloating in bagged fermented chili peppers was solved, the fermentation rate and product stability were improved, the use of preservatives was reduced, and a green and healthy fermentation effect was achieved.
Patent Information
- Application Number
- CN202511167524.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-20
- Publication Date
- 2025-11-18
AI Technical Summary
Bagged fermented chili peppers often experience problems such as bloating of bags and cans and leakage of juice during production, storage, transportation and sales. These problems are mainly caused by heterologous lactic acid bacteria and yeasts, especially Kazachstania yeasts. Improper use of existing preservatives can easily lead to food safety risks.
Lactobacillus plantarum SAAS-B-PCA-202043-6 was used to inhibit gas-producing yeast. By inoculating this strain in brine, low-salt live bacteria-type bagged fermented chili peppers were prepared, which improved the fermentation rate and stabilized product quality.
It effectively solves the bloating problem of bagged low-salt live bacteria fermented chili products, increases the fermentation rate, reduces the use of preservatives, maintains stable product quality, and has the advantages of being green and healthy.
Smart Images
Figure CN120966690A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of microbial fermentation technology, and particularly relates to the application of Lactobacillus plantarum SAAS-B-PCA-202043-6 in inhibiting gas-producing yeasts and its application in the preparation of bagged fermented chili peppers. Background Technology
[0002] Kimchi, a traditional fermented vegetable primarily fermented with lactic acid bacteria, has been passed down for thousands of years and is beloved for its unique characteristics of being "fragrant, tender, crisp, and refreshing." Kimchi fermentation falls under the category of "cold processing" of fresh vegetables. Under normal or low-temperature conditions, beneficial microorganisms continuously carry out metabolic activities. During this process, a series of complex physical, chemical, and biological reactions occur, generating mild flavor substances and aromatic components, giving kimchi its unique color, aroma, taste, and rich nutrition. This is the key to kimchi's enduring popularity throughout history.
[0003] With rising living standards and an increasingly fast pace of life, people's demand for convenient and quick fermented foods is growing. The industrialization of pickled vegetable production has made packaged or canned pickles a common appetizer on people's tables, giving rise to products such as pickled chili peppers, pickled cowpeas, pickled greens, and pickled radishes. Among them, chili peppers, as a typical representative of pickled vegetables, not only retain the shape, crispness, spiciness, and color of fresh chili peppers, but also possess the unique flavor, aroma, and health benefits produced by fermentation. Using polyethylene plastic bags as containers, fresh chili peppers are placed inside along with brine, inoculated with pure cultures, sealed, and fermented at room temperature to produce pickled chili peppers. Pickled chili peppers produced using this method meet hygiene standards, have a pure taste, can be produced year-round, and are extremely convenient to sell and consume, fully meeting consumers' demand for convenient food.
[0004] However, packaged pickled vegetables often experience problems such as bloating, can swelling, and juice leakage during production, storage, transportation, and sales. This severely shortens the product's shelf life, weakens the market competitiveness of producers, and causes unnecessary losses for both businesses and consumers. Taking pickled chili peppers as an example, the bloating is mainly caused by gas-producing bacteria such as heterotrophic lactic acid bacteria and yeasts, especially Kazachstania yeasts. In severe cases, this can lead to packaging bag rupture, leakage, and even food safety incidents. Currently, simply relying on the indiscriminate use of preservatives to solve the bloating problem is ineffective. Not only is it difficult to achieve the desired control effect, but it also easily leads to excessive preservative levels, increasing food safety risks. Summary of the Invention
[0005] In view of this, the purpose of this invention is to provide the application of *Lactobacillus plantarum* SAAS-B-PCA-202043-6 in inhibiting gas-producing yeasts and its application in the preparation of bagged fermented chili peppers. The *Lactobacillus plantarum* SAAS-B-PCA-202043-6 provided by this invention has a strong inhibitory effect on the main gas-producing yeasts in fermented vegetables. Applying it to chili pepper fermentation can increase the fermentation rate, shorten the fermentation cycle, effectively solve the gas problem in bagged low-salt live bacteria fermented chili pepper products, and stabilize product quality.
[0006] This invention provides the application of Lactiplantibacillus plantarum SAAS-B-PCA-202043-6 in inhibiting gas-producing yeasts.
[0007] Preferably, the gas-producing yeast includes *Saccharomyces cerevisiae*.
[0008] This invention provides the application of Lactobacillus plantarum SAAS-B-PCA-202043-6 in the preparation of bagged fermented chili peppers.
[0009] Preferably, the bagged fermented chili peppers are low-salt live bacteria fermented chili peppers.
[0010] This invention provides a method for preparing bagged low-salt live bacteria fermented chili peppers, comprising the following steps: pickling the chili peppers, mixing them with brine, packing them into a vacuum-sealed bag, and fermenting to obtain fermented chili peppers;
[0011] The brine is obtained by mixing spices, salt and water, boiling, cooling, and then inoculating with Lactobacillus plantarum SAAS-B-PCA-202043-6 fermentation broth.
[0012] The spices include star anise, Sichuan peppercorns, garlic, and ginger;
[0013] The inoculum size of the Lactobacillus plantarum SAAS-B-PCA-202043-6 fermentation broth is 1-3%; the fermentation temperature is 15-30℃; and the fermentation time is 14-30 days.
[0014] Preferably, the vacuum-sealed packaging bag has a specification of 250-1000g / bag.
[0015] Preferably, the viable cell concentration of the *Lactobacillus plantarum* SAAS-B-PCA-202043-6 fermentation broth is 10. 8 ~10 9 CFU / mL.
[0016] Preferably, the ratio of pickled chili peppers to brine is 1 kg: (1-2) L.
[0017] Preferably, the chili peppers are pickled using a pickling seasoning. Based on the weight of the chili peppers (100%), the pickling seasoning includes the following components: 3-5% salt, 1-3% white sugar, 0.3-0.8% sodium citrate, and 0.1-0.3% lactic acid. The pickling temperature is 18-22°C, and the pickling time is 2-4 hours.
[0018] Preferably, the brine contains 1% to 3% star anise, 0.04% to 0.06% Sichuan pepper, 0.08% to 0.12% garlic, 0.4% to 0.6% ginger, and 3% to 5% salt.
[0019] Compared with the prior art, the present invention has the following beneficial effects:
[0020] This invention screened a strain of *Lactobacillus plantarum* SAAS-B-PCA-202043-6 from the brine of secondary fermentation of sauerkraut. This strain inhibits gas-producing bacteria in fermented chili peppers. Applying it to chili pepper fermentation effectively solves the gas problem in packaged, low-salt, live-bacteria-based fermented chili pepper products, stabilizing product quality. Using sauerkraut fermentation bacteria as a control, the number of lactic acid bacteria, yeast, and total acid in chili peppers fermented with *Lactobacillus plantarum* SAAS-B-PCA-202043-6 were studied. The results showed that *Lactobacillus plantarum* has stronger adaptability in fermented sauerkraut, with high lactic acid bacteria content, high total acid production, and low yeast content, significantly reducing yeast count and effectively solving the gas problem in packaged fermented chili peppers. It has broad application prospects in fermented chili peppers. This invention not only provides a reference for developing lactic acid bacteria-fermented chili peppers but also offers a new approach for screening dedicated strains for fermented chili peppers.
[0021] Furthermore, compared to commercially available preservatives, the use of Lactobacillus plantarum SAAS-B-PCA-202043-6 in this invention not only has a good anti-bloating effect, but also reduces the use of preservatives, making it green and healthy. Moreover, the preparation method is simple, easy to operate, low in cost, and convenient and quick to apply, showing good prospects for promotion and application.
[0022] In addition, the *Lactobacillus plantarum* SAAS-B-PCA-202043-6 provided by this invention can also improve the fermentation rate, shorten the fermentation cycle, and maintain the stable color of fermented chili peppers. Attached Figure Description
[0023] Figure 1 Gas production during fermentation of low-salt bagged chili peppers using Lactobacillus plantarum SAAS-B-PCA-202043-6;
[0024] Figure 2 The growth curve of Lactobacillus plantarum SAAS-B-PCA-202043-6;
[0025] Figure 3The antibacterial activity of Lactobacillus plantarum SAAS-B-PCA-202043-6 against yeast;
[0026] Figure 4 The changes in yeast count during fermentation when only Kazachstania exigua radish juice was inoculated.
[0027] Biological Preservation Instructions
[0028] The Lactiplantibacillus plantarum SAAS-B-PCA-202043-6 provided by this invention is deposited at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC NO: 66512, deposit date of June 13, 2025, and deposit address of 5th Floor, Building 59, No. 100, Xianlie Middle Road, Guangzhou. Detailed Implementation
[0029] This invention provides the application of Lactiplantibacillus plantarum SAAS-B-PCA-202043-6 in inhibiting gas-producing yeasts.
[0030] In this invention, the *Lactobacillus plantarum* SAAS-B-PCA-202043-6 is isolated from the secondary fermentation water of sauerkraut; the colonies are milky white, with smooth surfaces and neat edges; it is a Gram-positive bacterium, and under a microscope, the cells are rod-shaped, arranged singly or in paired chains. The *Lactobacillus plantarum* exhibits good acid tolerance, salt tolerance, and antioxidant capacity. In this invention, the gas-producing yeast includes *Saccharomyces cerevisiae*.
[0031] The present invention also provides the application of Lactobacillus plantarum SAAS-B-PCA-202043-6 in the preparation of bagged fermented chili peppers.
[0032] In this invention, the bagged fermented chili peppers are low-salt, live bacteria fermented chili peppers. These low-salt, live bacteria fermented chili peppers have a low salt content and contain active lactic acid bacteria; no preservatives or sterilization are added during the preparation process.
[0033] This invention provides a method for preparing bagged low-salt live bacteria fermented chili peppers, comprising the following steps: pickling the chili peppers, mixing them with brine, packing them into a vacuum-sealed bag, and fermenting to obtain fermented chili peppers;
[0034] In this invention, there is no particular limitation on the type of chili peppers used; any chili pepper variety commonly used in the art can be employed, including but not limited to one or more combinations of Red Erjingtiao, Red Millet Chili, Beauty Chili, Wild Chili, and Lantern Chili. Fresh chili peppers are preferred in this invention. Before use, the inedible parts of the chili peppers are preferably removed, and they are thoroughly cleaned and set aside. The chili peppers are then pickled using a pickling compound. Based on the weight of the chili peppers (100%), the pickling compound comprises the following components: 3-5% salt, 1-3% white sugar, 0.3-0.8% sodium citrate, and 0.1-0.3% lactic acid; preferably, it comprises: 3.5-4.5% salt, 1.5-2.5% white sugar, 0.4-0.7% sodium citrate, and 0.15-0.25% lactic acid; and even more preferably, it comprises: 4% salt, 2% white sugar, 0.5% sodium citrate, and 0.2% lactic acid. In this invention, the marinating temperature is preferably 18-22°C, more preferably 19-31°C, and even more preferably 20°C; the marinating time is preferably 2-4 hours, and even more preferably 3 hours.
[0035] In this invention, the brine is obtained by mixing spices, salt, and water, boiling, cooling, and then inoculating with *Lactobacillus plantarum* SAAS-B-PCA-202043-6 fermentation broth. In this invention, the spices include star anise, Sichuan pepper, garlic, and ginger; preferably, the brine contains 1%–3% star anise, 0.04%–0.06% Sichuan pepper, 0.08%–0.12% garlic, 0.4%–0.6% ginger, and 3%–5% salt; more preferably, it contains 1.5%–2.5% star anise, 0.045%–0.055% Sichuan pepper, 0.09%–0.11% garlic, 0.45%–0.55% ginger, and 3.5%–4.5% salt; even more preferably, it contains 2% star anise, 0.05% Sichuan pepper, 0.1% garlic, 0.5% ginger, and 4% salt.
[0036] In this invention, the inoculation amount of the *Lactobacillus plantarum* SAAS-B-PCA-202043-6 fermentation broth is 1% to 3%, preferably 1.5% to 2.5%, and more preferably 2%; the viable cell concentration of the inoculated *Lactobacillus plantarum* SAAS-B-PCA-202043-6 fermentation broth is preferably 10. 8 ~10 9 CFU / mL.
[0037] In this invention, chili peppers are pickled, mixed with brine, and placed in vacuum-sealed bags for fermentation to obtain fermented chili peppers. The preferred ratio of pickled chili peppers to brine is 1 kg:(1-2) L, more preferably 1 kg:(1.2-1.8) L; the vacuum-sealed bags are 250-1000 g / bag, specifically 250 g, 300 g, 500 g, and 1000 g; the fermentation temperature is preferably 15-30°C, more preferably 18-25°C; and the fermentation time is preferably 14-30 days, more preferably 15-20 days.
[0038] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.
[0039] Material
[0040] Secondary fermentation water of pickled cabbage: collected from a food company in Sichuan;
[0041] Standard fermentation agent: Shangchuan pickled vegetable fermentation bacteria, purchased from Guangdong Shunde Shangchuan Biotechnology Co., Ltd.;
[0042] Kazachstania exigua was isolated from fermented mustard greens by the Fruit and Vegetable Fermentation Food Research Group of the Institute of Agricultural Product Processing (Institute of Food and Nutrition Health, Sichuan Academy of Agricultural Sciences).
[0043] MRS agar medium: peptone 10.0 g / L, beef extract 5.0 g / L, yeast extract 4.0 g / L, glucose 20.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, triammonium citrate 2.0 g / L, sodium acetate 5.0 g / L, magnesium sulfate 0.2 g / L, manganese sulfate 0.05 g / L, agar 15.0 g / L, Tween 80 1.0 g / L; pH 7.0 ± 0.2.
[0044] MRS meat culture medium: peptone 10.0 g / L, beef extract 5.0 g / L, yeast extract 4.0 g / L, glucose 20.0 g / L, dipotassium hydrogen phosphate 2.0 g / L, triammonium citrate 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.
[0045] Sterilized radish juice: Take an appropriate amount of red radish, remove the roots and leaves, crush it with a juicer, filter it through 4 layers of gauze, boil the filtrate for 30 seconds to remove the surface film, obtain radish juice, sterilize it at 115℃ for 15 minutes, and set aside.
[0046] Example 1
[0047] Detection of the anti-flatulence ability of Lactobacillus plantarum SAAS-B-PCA-202043-6
[0048] Take an appropriate amount of wild chili peppers, remove inedible parts, wash and drain the surface water. Inoculate *Lactobacillus plantarum* SAAS-B-PCA-202043-6 at a 1% inoculation rate into a package containing wild chili peppers and water at a ratio of 200g:200mL (containing 4% salt). Vacuum seal the package and ferment at 25℃ for 20 days, observing the gas expansion of the packaged peppers. The control group without added bacteria was treated the same as with *Lactobacillus plantarum*, except that no additional bacterial strain was added. Results are as follows: Figure 1 As shown, after 20 days of fermentation, the bagged fermented peppers with added SAAS-B-PCA-202043-6 only produced weak small bubbles, while the bagged fermented peppers without added bacteria produced a large amount of gas, causing the height of the bagged peppers to rise from 3cm to 6cm. Therefore, it is determined that SAAS-B-PCA-202043-6 has a significant ability to inhibit gas-producing bacteria.
[0049] Gas production and film formation
[0050] Activated SAAS-B-PCA-202043-6 was inoculated into test tubes containing Durham tubes of MRS liquid culture medium and incubated at 36℃ for 24 hours. The presence of air bubbles in the Durham tubes and a white film on the surface of the liquid culture medium was observed. The results showed that SAAS-B-PCA-202043-6 neither produces gas nor film, making it an excellent lactic acid bacterium for kimchi fermentation.
[0051] Growth curve of Lactobacillus plantarum SAAS-B-PCA-202043-6
[0052] Activated *Lactobacillus plantarum* was inoculated into MRS vegetative cells at a 1% inoculum and cultured at 36℃ for 24 hours. Samples were taken every 4 hours, and the OD value of the fermentation broth was measured at an absorbance of 600 nm. The results are as follows: Figure 2 As shown, lactic acid bacteria enter the logarithmic phase after 4 hours and the stationary phase after 16 hours.
[0053] Example 2
[0054] Lactobacillus plantarum's inhibitory effect on Kazachstania exigua, the main flatulence-causing yeast in fermented vegetables
[0055] Activated *Lactobacillus plantarum* and *Kazachstania exigua* were inoculated into sterile radish juice at a 1% inoculum concentration. A control group inoculated only with 1% *Kazachstania exigua* was also included. The mixture was incubated at 30°C for 5 days. Fermented radish juice was collected on days 0, 1, 3, and 5, and the numbers of lactic acid bacteria and yeast were counted. The results are as follows: Figure 3 and Figure 4 As shown. From Figure 3 It can be seen that *Lactobacillus plantarum* showed an increasing trend during fermentation, reaching a colony count of 9.40 lg (CFU / mL) after 3 days of fermentation, while the yeast count showed a slow decreasing trend, from 5.36 lg (CFU / mL) at the beginning of fermentation to 4.53 lg (CFU / mL) after 5 days of fermentation, a decrease of 0.83 lg (CFU / mL). Figure 4 It was found that when radish juice inoculated only with Kazachstania exigua was used, the yeast count showed an increasing trend during fermentation, reaching 7.27 CFU / mL after 5 days of fermentation. Therefore, this *Lactobacillus plantarum* strain has a strong inhibitory effect on Kazachstania exigua, the main flatulence-causing yeast in fermented vegetables.
[0056] Application Experiment Example 1
[0057] Fermented peppers containing Lactobacillus plantarum
[0058] (1) Pre-treatment of raw materials: Remove the inedible parts of fresh chili peppers, wash them clean, and set aside;
[0059] (2) Raw material pickling: Add 4% salt, 2% white sugar, 0.5% sodium citrate and 0.2% lactic acid according to the weight of chili peppers, mix well and pickle at 20℃ for 3 hours.
[0060] (3) Preparation of brine: Take 0.2% star anise, 0.05% Sichuan pepper, 0.1% garlic and 0.5% ginger, put them into purified water, add 4% salt by volume, heat to boiling and then cool. Cool the boiled brine to room temperature, then add 2% Lactobacillus plantarum fermentation liquid (10 9 (CFU / mL) was used to obtain brine.
[0061] (4) Fermentation by sealing: The products from steps (2) and (3) are packed into a packaging bag in a 1:1 ratio, vacuum sealed, and packaged in a 500g bag at 20℃ for 15 days. The total acid content reaches more than 0.8%, and fermented chili peppers are obtained.
[0062] Comparative Example 1
[0063] (1) Pre-treatment of raw materials: Remove the inedible parts of fresh chili peppers, wash them clean, and set aside;
[0064] (2) Marinating of raw materials: Add 4% salt, 2% white sugar, 0.5% sodium citrate and 0.2% lactic acid according to the weight ratio, mix well and marinate at 20℃ for 3 hours.
[0065] (3) Preparation of brine: Take 0.2% star anise, 0.05% Sichuan pepper, 0.1% garlic and 0.5% ginger, put them into pure water, add 4% salt according to the water volume, heat to boiling and then cool. Cool the boiled brine to room temperature, and then add 2% conventional fermentation agent to obtain brine.
[0066] (4) Fermentation by sealing: The products from steps (2) and (3) are packed into a packaging bag in a 1:1 ratio, vacuum sealed, and packaged in a 500g bag at 20℃ for 15 days. The total acid content reaches more than 0.8%, and fermented chili peppers are obtained.
[0067] The content of lactic acid bacteria, yeast, and total acid in fermented peppers was determined in two groups: Experimental Example 1 and Comparative Example 1.
[0068] The determination of lactic acid bacteria content should refer to GB 4789.35-2023;
[0069] The determination of yeast content should refer to GB4789.15-2016;
[0070] The determination of total acidity shall be performed by potentiometric titration, Method II, in GB12456-2021.
[0071] The results showed that *Lactobacillus plantarum* could grow well in fermented peppers, with lactic acid bacteria reaching 4.5 × 10⁻⁶. 9 The CFU / g of this product is superior to the 2.7×10⁻⁶ lactic acid bacteria content of conventional starter cultures. 7 The CFU / g concentrations were significantly different (P<0.05); the yeast content in the fermented peppers by *Lactobacillus plantarum* was 1.5×10⁻⁶. 4 CFU / g, while the yeast content of conventional starter cultures is 3.4 × 10⁻⁶. 7 The total acidity of chili peppers fermented with *Lactobacillus plantarum* was 1.03%, while that of conventional starter cultures was 0.85%. This demonstrates that *Lactobacillus plantarum* exhibits stronger adaptability in fermented chili peppers, with higher lactic acid bacteria content, higher total acidity, and lower yeast content. Therefore, adding *Lactobacillus plantarum* to fermented chili peppers can effectively solve the problem of bloating in packaging of low-salt live bacteria fermented chili peppers, and has broad application prospects in chili pepper fermentation.
[0072] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. Application of Lactobacillus plantarum (SAAS-B-PCA-202043-6) in inhibiting gas-producing yeasts.
2. The application according to claim 1, characterized in that, The gas-producing yeast includes oligosporus Kazakhstan yeast.
3. Application of Lactobacillus plantarum SAAS-B-PCA-202043-6 in the preparation of bagged fermented chili peppers.
4. The application according to claim 3, characterized in that, The bagged fermented chili peppers are low-salt live bacteria fermented chili peppers.
5. A method for preparing bagged low-salt live bacteria fermented chili peppers, characterized in that, Includes the following steps: After pickling the chili peppers, they are mixed with brine, packed into vacuum-sealed bags, and fermented to obtain fermented chili peppers. The brine is obtained by mixing spices, salt and water, boiling, cooling, and then inoculating with Lactobacillus plantarum SAAS-B-PCA-202043-6 fermentation broth. The spices include star anise, Sichuan peppercorns, garlic, and ginger; The inoculum size of the Lactobacillus plantarum SAAS-B-PCA-202043-6 fermentation broth is 1-3%; the fermentation temperature is 15-30℃; and the fermentation time is 14-30 days.
6. The preparation method according to claim 5, characterized in that, The vacuum-sealed packaging bag has a specification of 250-1000g / bag.
7. The preparation method according to claim 5, characterized in that, The viable cell concentration of the *Lactobacillus plantarum* SAAS-B-PCA-202043-6 fermentation broth was 10. 8 ~10 9 CFU / mL.
8. The preparation method according to claim 5, characterized in that, The ratio of pickled chili peppers to brine is 1 kg: (1-2) L.
9. The preparation method according to claim 5, characterized in that, The chili peppers are pickled using a pickling seasoning. Based on the weight of the chili peppers (100%), the pickling seasoning includes the following components: 3-5% salt, 1-3% white sugar, 0.3-0.8% sodium citrate, and 0.1-0.3% lactic acid. The pickling temperature is 18-22°C, and the pickling time is 2-4 hours.
10. The preparation method according to claim 5, characterized in that, The brine contains 1% to 3% star anise, 0.04% to 0.06% Sichuan pepper, 0.08% to 0.12% garlic, 0.4% to 0.6% ginger, and 3% to 5% salt.