Lactobacillus plantarum and debaryomyces hansenii as well as application of lactobacillus plantarum and debaryomyces hansenii in preparation of preserved vegetables through fermentation
By introducing specific strain fermentation technology and using pickled cabbage leaves to make dried plums, the problems of waste of pickled cabbage resources and unstable quality are solved, and the efficient utilization of the pickled cabbage industry and product quality improvement are achieved.
Patent Information
- Application Number
- CN202510492477.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-18
- Publication Date
- 2025-07-18
AI Technical Summary
The by-product of traditional pickled cabbage production has not been effectively utilized, resources are seriously wasted, the natural fermentation quality is unstable, and there is a lack of industrial support.
Lactobacillus plantarum (CGMCC NO.26826) and Debaryomyces hansenii (CGMCC NO.33845) were used as targeted strains. Pulp leaf was made into dried plums through a specific fermentation process, and auxiliary materials were added to fermented and sterilized.
It has improved the utilization rate of pickled vegetables, improved the flavor and nutritional value of dried plums, achieved stability of the fermentation process and consistency of products, and has economic, environmental and social benefits.
Smart Images
Figure CN120330099A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the field of microorganisms and relates to the application of Lactobacillus plantarum and Debaryomyces hansenii in the fermentation of making preserved mustard greens. Background Art
[0002] After 120 years of development, Fuling mustard tuber has become one of the world's three famous pickled vegetables together with European pickled cucumbers and German sweet and sour cabbages, and is well-known both at home and abroad. It has developed into a dominant characteristic pillar industry in the agricultural and rural economy of Fuling District and even Chongqing City, with large production and sales scale, high brand awareness, and strong radiation and driving ability. It has accumulated and formed unique development advantages in variety breeding, planting technology, production and processing, brand culture, etc. In 2021, the harvested area of green vegetable heads in Fuling was 729,500 mu, and the total output was 1.626 million tons. However, only the green vegetable heads are used in the production of mustard tuber, and most of the by-products (mainly leaves) are discarded in the fields without forming industrialization, resulting in waste of resources.
[0003] Therefore, it has become an urgent task to vigorously strengthen the comprehensive utilization of mustard tuber by-products and key technology research, develop new products using mustard tuber by-products, and improve the resource utilization rate of mustard tuber. This product uses mustard tuber leaves as raw materials and ferments to make "preserved mustard greens" by adding appropriate strains. It not only develops high-quality, green and healthy products, but also improves the resource utilization rate of mustard tuber, increases its added value, and has good economic and social benefits. Summary of the Invention
[0004] In view of this, one of the purposes of the present invention is to provide Lactobacillus plantarum and Debaryomyces hansenii, and the second purpose is to provide the application of Lactobacillus plantarum and Debaryomyces hansenii and their mixture in the fermentation of making preserved mustard greens.
[0005] To achieve the above object, the present invention provides the following technical solutions:
[0006] The present invention provides a Lactobacillus plantarum, classified and named as: Lactobacillus plantarum, deposit number: CGMCC NO.26826;
[0007] The present invention provides a Debaryomyces hansenii, classified and named as: Debaryomyces hansenii, deposit number: CGMCC NO.33845;
[0008] It also provides a group of microbiomes including the above-mentioned Lactobacillus plantarum and the above-mentioned Debaryomyces hansenii;
[0009] Furthermore, the application of Lactobacillus plantarum in the fermentation of making preserved mustard greens;
[0010] Furthermore, the application of the above-mentioned Debaryomyces hansenii in the fermentation of making preserved mustard greens;
[0011] Further, the application of the microbiome in the fermentation of pickled mustard tuber leaves to make preserved mustard greens;
[0012] Further, a method for fermenting pickled mustard tuber leaves to make preserved mustard greens, the steps are as follows:
[0013] (1) Pretreatment of pickled mustard tuber leaves: The picked pickled mustard tuber leaves are sun-dried until wilted, washed and drained, cut into sections, and then sun-dried until wilted for standby;
[0014] (2) Preparation of bacterial liquid: The strain of claim 1 or 2 is activated on a solid medium, a single colony is picked and inoculated into a liquid medium, cultured at 30 °C for 48 hours, centrifuged at 4500 r / min for 10 minutes, the supernatant is discarded, washed and centrifuged twice with sterile normal saline, and finally added with sterile normal saline with a volume of 1 / 2 of the original medium volume to prepare a bacterial suspension of 10 7 ~10 8 CFU / mL as the fermentation agent;
[0015] (3) Inoculation and fermentation: Add 0.5% Chinese prickly ash powder, 3% chili powder, 0.2% thirteen-spice powder, and 0.5% high-proof liquor according to the mass of pickled mustard tuber leaves, inoculate the fermentation agent prepared in step (2), mix well and knead, then pack into a jar and compact, and seal and ferment in an inverted water-sealed jar;
[0016] (4) Packaging treatment: Take out after 6 months of fermentation, carry out vacuum packaging and implement pasteurization;
[0017] Preferably, in step (2), the Lactobacillus plantarum is activated and cultured with MRS medium, and the Debaryomyces hansenii is activated and cultured with YPD medium;
[0018] Preferably, in step (3), the mass of the inoculated fermentation agent is 1%, and the fermentation agent is any one of a Lactobacillus plantarum fermentation agent, a Debaryomyces hansenii fermentation agent, or a fermentation agent with a 1:1 mixture of Lactobacillus plantarum and Debaryomyces hansenii.
[0019] The beneficial effects of the present invention are as follows:
[0020] 1. Efficient utilization of resources
[0021] Solve the problem of waste of by-products. By using the pickled mustard tuber leaves that are traditionally discarded as raw materials, the resource utilization rate of the Fuling pickled mustard tuber industry is greatly improved, agricultural waste is reduced, and it conforms to the concept of green and sustainable development. Industrialization potential: Convert the pickled mustard tuber leaves that originally had no economic value into high-value-added products (preserved mustard greens), creating a new source of economic income.
[0022] 2. Improve product quality
[0023] The pickled mustard greens flavored enhanced by fermenting with the strains of the present invention endows the product with a stronger sour and sweet taste, improved nutritional value, more sufficient protein decomposition, and enhanced umami and nutrition. The pickled mustard greens product prepared by the present invention has better color, texture and flavor compared with the natural fermentation method.
[0024] 3. Process standardization and controllability
[0025] By using the specific strains of the present invention (Lactobacillus plantarum ZS and Debaryomyces hansenii JYY2) to replace the uncontrollable microbial communities in traditional natural fermentation, the stability of the fermentation process and the consistency of the product are ensured.
[0026] 4. Synergistic effect of strains
[0027] Advantages of combined fermentation: When Lactobacillus plantarum and Debaryomyces hansenii are used in combination (ZS+JYY2 group), the fermentation efficiency is synergistically improved.
[0028] By innovatively introducing Lactobacillus plantarum and Debaryomyces hansenii, the present invention not only solves the problem of waste of mustard green leaves, but also significantly improves the sensory quality and nutritional value of pickled mustard greens. At the same time, the standardization of the production process is achieved, with significant economic, environmental and social benefits.
[0029] Other advantages, objectives and features of the present invention will be described to some extent in the subsequent specification, and to some extent, will be obvious to those skilled in the art based on the study of the following text, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following specification. Brief description of the drawings
[0030] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be described in preferred detail below in conjunction with the drawings, where:
[0031] Figure 1 is pickled mustard greens by natural fermentation;
[0032] Figure 2 is pickled mustard greens fermented by inoculating JYY2 (Debaryomyces hansenii);
[0033] Figure 3 is pickled mustard greens fermented by inoculating ZS (Lactobacillus plantarum);
[0034] Figure 4 is pickled mustard greens fermented by inoculating ZS+JYY2 (Debaryomyces hansenii + Lactobacillus plantarum). Detailed implementation manners
[0035] The following uses specific specific examples to illustrate the implementation manners of the present invention. Those skilled in the art can easily understand the other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the diagrams provided in the following embodiments only schematically illustrate the basic concept of the present invention. Without conflict, the following embodiments and the features in the embodiments can be combined with each other.
[0036] Among them, the attached drawings are only for illustrative purposes, showing only schematic diagrams, not physical diagrams, and should not be construed as a limitation to the present invention; in order to better illustrate the embodiments of the present invention, some components in the attached drawings will be omitted, enlarged or reduced, which does not represent the size of the actual product; for those skilled in the art, it is understandable that some well-known structures and their descriptions in the attached drawings may be omitted.
[0037] In the attached drawings of the embodiments of the present invention, the same or similar reference numerals correspond to the same or similar components; in the description of the present invention, it should be understood that if there are terms such as "upper", "lower", "left", "right", "front", "rear", etc. indicating the orientation or position relationship, it is based on the orientation or position relationship shown in the attached drawings. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation. Therefore, the terms describing the position relationship in the attached drawings are only for illustrative purposes and should not be construed as a limitation to the present invention. For those of ordinary skill in the art, the specific meanings of the above terms can be understood according to specific circumstances.
[0038] Example 1
[0039] Natural fermentation method:
[0040] (1) Pretreatment of mustard leaves: After the mustard leaf raw materials are picked, they are sun-dried until wilted, then washed clean and drained of water, cut into vegetable segments, and sun-dried until wilted, with a yield of about 30%; reserve for later use.
[0041] (2) Natural fermentation to make "preserved vegetables" from mustard leaves: Add 0.5% of ground pepper, 3% of chili powder, 0.2% of thirteen-spice powder, and 0.5% of high-proof liquor according to the mass of mustard leaves, without adding a fermentation agent (natural fermentation), mix well and knead, then put it into an inverted earthenware jar and compact it, and then seal the inverted earthenware jar with water and ferment it sealed.
[0042] (3) Packaging: After fermenting for more than 6 months, take it out, vacuum package it and perform pasteurization.
[0043] Example 2
[0044] JYY2 (Debaryomyces hansenii) fermentation method:
[0045] (1) Pretreatment of mustard green leaves: After picking the raw materials of mustard green leaves, let them wilt in the sun, then wash them clean and drain the water, cut them into sections, and dry them until wilted. The yield is about 30%; reserve for later use.
[0046] (2) Preparation of bacterial liquid: The JYY2 (Debaryomyces hansenii, preservation number: CGMCC NO. 33845) strain stored in a -20°C refrigerator was activated on YPD medium. Then, a single colony was picked and cultured in YPD liquid medium at 30°C for 48 h, centrifuged at 4500 r / min for 10 min, the supernatant was discarded, washed twice by centrifugation with sterile normal saline, the supernatant was discarded, and then sterile normal saline with a volume of 1 / 2 of the original medium was added to prepare a bacterial suspension (concentration 10 7 ~10 8 CFU / mL), and the JYY2 starter was prepared.
[0047] (3) Inoculation and fermentation of mustard green leaves into "preserved vegetables": Add 0.5% of Chinese prickly ash powder, 3% of chili powder, 0.2% of thirteen-spice powder, and 0.5% of high-proof liquor according to the mass of mustard green leaves. Add the JYY2 starter at an inoculation amount of 1%, mix well and knead, then put it into an inverted earthenware jar and compact it, and then seal the inverted earthenware jar with water and ferment it sealed.
[0048] (4) Packaging: After fermenting for more than 6 months, take it out, vacuum pack it and perform pasteurization.
[0049] Example 3
[0050] Fermentation method with ZS (Lactobacillus plantarum):
[0051] (1) Pretreatment of mustard green leaves: After picking the raw materials of mustard green leaves, let them wilt in the sun, then wash them clean and drain the water, cut them into sections, and dry them until wilted. The yield is about 30%; reserve for later use.
[0052] (2) Preparation of bacterial liquid: The ZS (Lactobacillus plantarum, preservation number: CGMCC NO. 26826) strain stored in a -20°C refrigerator was activated on MRS solid medium. Then, a single colony was picked and cultured in MRS broth medium at 30°C for 48 h, centrifuged at 4500 r / min for 10 min, the supernatant was discarded, washed twice by centrifugation with sterile normal saline, the supernatant was discarded, and then sterile normal saline with a volume of 1 / 2 of the original medium was added to prepare a bacterial suspension (concentration 10 7 ~10 8 CFU / mL), and the ZS starter was prepared.
[0053] (3) Inoculation and fermentation of pickled mustard tuber leaves into "preserved vegetables": Add 0.5% of Chinese prickly ash powder, 3% of chili powder, 0.2% of thirteen-spice powder, and 0.5% of high-proof liquor according to the mass of pickled mustard tuber leaves. Add the ZS fermenting agent at an inoculation amount of 1%, mix well and knead, then put it into an inverted earthenware jar and compact it, and then seal the inverted earthenware jar with water and ferment it sealed.
[0054] (4) Packaging: After fermentation for more than 6 months, take it out, vacuum package it and perform pasteurization.
[0055] Example 4
[0056] Fermentation method of ZS + JYY2 (Debaryomyces hansenii + Lactobacillus plantarum):
[0057] (1) Pretreatment of pickled mustard tuber leaves: After picking the pickled mustard tuber leaf raw materials, dry them in the sun until withered, then wash them clean and drain the water, cut them into sections, and dry them in the sun until withered. The yield is about 30%; reserve for use.
[0058] (2) Preparation of bacterial suspension: The ZS (Lactobacillus plantarum, preservation number: CGMCC NO.26826) and JYY2 (Debaryomyces hansenii, preservation number: CGMCC NO.33845) strains stored in a -20°C refrigerator were activated on MRS and YPD media, then single colonies were picked and cultured in MRS broth and YPD liquid media at 30°C for 48 h, centrifuged at 4500 r / min for 10 min, the supernatant was discarded, washed by centrifugation with sterile normal saline twice, the supernatant was discarded, and then sterile normal saline with a volume of 1 / 2 of the original medium was added to prepare a bacterial suspension (concentration 10 7 ~10 8 CFU / mL), and the ZS and J1 bacterial suspensions were mixed evenly at a ratio of 1:1 to prepare a mixed fermenting agent.
[0059] (3) Inoculation and fermentation of pickled mustard tuber leaves into "preserved vegetables": Add 0.5% of Chinese prickly ash powder, 3% of chili powder, 0.2% of thirteen-spice powder, and 0.5% of high-proof liquor according to the mass of pickled mustard tuber leaves. Add the mixed fermenting agent at an inoculation amount of 1%, mix well and knead, then put it into an inverted earthenware jar and compact it, and then seal the inverted earthenware jar with water and ferment it sealed.
[0060] (4) Packaging: After fermentation for more than 6 months, take it out, vacuum package it and perform pasteurization.
[0061] Example 5
[0062] Evaluate the pickled mustard tuber fermentation methods in Examples 1-4.
[0063] Table 1 Titratable acid, reducing sugar, amino acid, nitrite and sensory evaluation of pickled mustard tuber leaves prepared by different preparation methods
[0064]
[0065] From the comparison results in Table 1, it can be seen that the titratable acid content of the mustard leaves inoculated with J1 (Debaryomyces hansenii) is higher (5.57±0.00d), the reducing sugar content is higher (3.33±0.01d), the amino acid content is higher (1052.20±34.43c), and the sensory evaluation is better (83.00±3.02c). Therefore, the quality of the mustard leaves prepared by fermentation with the inoculation of JYY2 (Debaryomyces hansenii) is the best.
[0066] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the spirit and scope of the present technical solution, and they should all be covered by the scope of the claims of the present invention.
Claims
1. Lactobacillus plantarum, characterized in that: The preservation number of the Lactobacillus plantarum is: CGMCC NO.26826.
2. Debaryomyces hansenii, characterized in that: The preservation number of the Debaryomyces hansenii is: CGMCC NO.33845.
3. The microbiome comprises the Lactobacillus plantarum according to claim 1 and the Debaryomyces hansenii according to claim 2.
4. Use of the Lactobacillus plantarum according to claim 1 in the fermentation for making preserved mustard green.
5. Use of the Debaryomyces hansenii according to claim 2 in the fermentation for making preserved mustard green.
6. Use of the microbiome according to claim 3 in the fermentation for making preserved mustard green.
7. A method for fermenting and making preserved vegetables with plum, characterized in that, The steps are as follows: (1) Pretreatment of mustard green leaves: The picked mustard green leaves are sun-dried until wilted, cut into sections after washing and draining, and then sun-dried again until wilted for standby; (2) Preparation of bacterial suspension: Activate the strain of Claim 1 or 2 on a solid medium, pick a single colony and inoculate it into a liquid medium, culture at 30 °C for 48 hours, centrifuge at 4500 r / min for 10 minutes, discard the supernatant, wash and centrifuge twice with sterile normal saline, and finally add sterile normal saline with a volume of 1 / 2 of the original medium to prepare a bacterial suspension of 10 7 ~10 8 CFU / mL as the starter; (3) Inoculation and fermentation: Add 0.5% Chinese prickly ash powder, 3% chili powder, 0.2% thirteen-spice powder, and 0.5% high-proof liquor according to the mass of the mustard green leaves, inoculate the fermenting agent prepared in step (2), mix well and knead, then pack into a jar and compact, and seal and ferment with the water seal upside down; (4) Packaging treatment: Take out after 6 months of fermentation, carry out vacuum packaging and implement pasteurization.
8. The method for fermenting and making preserved mustard greens according to claim 7, characterized in that: In step (2), the Lactobacillus plantarum uses MRS medium, and the Debaryomyces hansenii uses YPD medium.
9. The method for fermenting and making preserved mustard greens according to claim 7, characterized in that: In step (3), the mass of the inoculated fermenting agent is 1%, and the fermenting agent is any one of a Lactobacillus plantarum fermenting agent, a Debaryomyces hansenii fermenting agent, or a fermenting agent with a 1:1 mixture of Lactobacillus plantarum and Debaryomyces hansenii.