Method for preparing rose beverage through fermentation of lactobacillus paracasei
Through an innovative process, the use of Lactobacillus paracasei extracted from Northeast sauerkraut to ferment a rose beverage, solving the problems of complex processes and allergic risks in the existing process, realizing the unique taste and flavor of the beverage, with potential health benefits and efficient anticorrosion effects.
Patent Information
- Application Number
- CN202510262772.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-06
- Publication Date
- 2025-05-30
AI Technical Summary
The existing process of extracting Lactobacillus paracasei from Northeast sauerkraut is complex, and it is greatly affected by the quality of raw materials, and there is a potential risk of allergies. It is rare to use this strain for fermentation.
An innovative process is adopted to make rose beverages by fermenting sauerkraut sample preparation, homogenization, inoculation and culture, colony isolation and identification, Lactobacillus paracasei preservation, rose petal selection and treatment, mixing and leaching, high-temperature sterilization, fermentation and secondary fermentation.
By optimizing the fermentation process and formula design, the unique taste and flavor of the beverage is improved, the fragrance of roses and the sweet and sour taste of Lactobacillus paracasei are retained, bringing consumers a pleasant drinking experience, and significantly improving the yield of bacterial sterilization, producing food preservatives with efficient preservative effects.
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Figure CN120052480A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of beverage processing, and more specifically, to a method for producing rose beverage by fermenting with Lactobacillus paracasei. Background Art
[0002] Extracting Lactobacillus paracasei from Northeast sauerkraut is a biotechnological method with practical application value. Lactobacillus paracasei is an important probiotic bacterium, widely present in fermented foods such as sauerkraut and pickles. In Northeast sauerkraut, Lactobacillus paracasei is one of the main fermentation strains, endowing sauerkraut with a unique taste and flavor through fermentation. The process of extracting Lactobacillus paracasei includes steps such as sampling from sauerkraut, isolation and purification, morphological observation, physiological and biochemical identification, etc. Through a series of culturing and screening, Lactobacillus paracasei with relatively high purity can be obtained. These strains have broad application prospects in the food industry, such as being used as fermenting agents, preservatives or probiotic preparations, etc. Existing technologies have also shown that certain Lactobacillus paracasei (such as HD1.7) can produce bacteriocins with broad-spectrum antibacterial effects, such as Paracin1.7. Therefore, extracting Lactobacillus paracasei from Northeast sauerkraut not only helps to deeply understand the microbial mechanism of sauerkraut fermentation, but also provides high-quality probiotic resources for the food industry.
[0003] Rose (Rosa rugosa Thunb): It is a general term for various plants and cultivated flowers of the Rosaceae family and the genus Rosa. Rose petals can be made into pie fillings, rose wine, rose syrup, and can be used to make tea after drying. The fruits contain rich vitamin C, malic acid, carotene, etc.; fresh flowers can be steamed to produce aromatic oils for food and cosmetic use. Rose flowers contain various trace elements and have a high content of vitamin C. Rose flowers can be used to make various pastries, such as rose candies, rose cakes, rose tea, rose wine, rose pickles, rose paste, etc. Rose flowers can be directly eaten in some areas of Europe. Rose roots can be cooked and eaten, and rose roots can be used to make wine.
[0004] Although the method of fermenting with lactic acid bacteria has been widely recognized, the method of fermenting with Lactobacillus paracasei extracted from Northeast sauerkraut is not common.
[0005] The existing processes for extracting Lactobacillus paracasei from sauerkraut have the following problems: The extraction process is complex. Extracting Lactobacillus paracasei from sauerkraut requires a series of complex steps and has relatively high requirements for technology and equipment. Affected by the quality of raw materials: The quality of sauerkraut directly affects the quantity and activity of the extracted Lactobacillus paracasei. If the sauerkraut is contaminated during the fermentation process or the fermentation conditions are not good, it may lead to a decline in the quality of the extracted Lactobacillus paracasei. Potential allergy risk: Although Lactobacillus paracasei has high safety, for some people with allergic constitutions, there may still be an allergy risk if the quality of Lactobacillus paracasei is not good.
[0006] Therefore, whether it is possible to provide a method for producing rose beverages by fermenting Lactobacillus paracasei to improve the above technical deficiencies is an urgent problem to be solved by those skilled in the art. Summary of the Invention
[0007] In view of this, the present invention provides a method for producing rose beverages by fermenting Lactobacillus paracasei. It is committed to developing a new method to improve the taste of beverages and enhance their nutritional value through the fermentation of Lactobacillus paracasei. The core lies in exploring an innovative process for extracting Lactobacillus paracasei from Northeast sauerkraut.
[0008] To achieve the above object, the present invention adopts the following technical solutions:
[0009] A method for producing rose beverages by fermenting Lactobacillus paracasei, comprising the following steps:
[0010] (1) Preparation of sauerkraut samples;
[0011] (2) Sample homogenization: Add the cut small pieces of sauerkraut to water; after mixing and stirring, mash; then, stir the mashed mixture to make it evenly dispersed; perform gradient dilution on the mixture, and the diluent is sterile normal saline;
[0012] (3) Inoculation and cultivation: The culture medium is sterilized by high pressure, and then, the homogenate of the mixture diluted in step (2) is inoculated onto the culture medium for cultivation;
[0013] (4) Colony isolation and identification to screen Lactobacillus paracasei;
[0014] (5) Preservation of Lactobacillus paracasei;
[0015] (6) Selection and treatment of rose petals: First, remove impurities and select flawless rose petals, then, soak and wash them multiple times, and then perform crushing treatment to obtain crushed petals;
[0016] (7) Preparation of sugar solution: Slowly add granulated sugar to purified water until it is completely dissolved to form a uniform sugar solution;
[0017] (8) Mixed extraction: Add purified water, crushed petals and sugar solution to the fermentation tank, after heating, add the remaining purified water, and perform mixed extraction to obtain the material;
[0018] (9) High-temperature sterilization;
[0019] (10) Fermentation: After sterilization is completed, cool the material, add Lactobacillus paracasei, and maintain constant-temperature fermentation culture;
[0020] (11) Secondary fermentation;
[0021] (12) Finished product: Sterilize and fill to obtain rose beverage.
[0022] Preferably, step (1) is specifically as follows: Collect fresh Northeast pickled vegetables, first drain the pickled vegetables to remove the water, then remove the inedible parts, and cut the pickled vegetables into small pieces.
[0023] Preferably, step (2) is specifically as follows: Put the cut small pieces of pickled vegetables into a blender and add clear water enough to cover the top of the pickled vegetables; After mixing and stirring for 10 - 20 min, start the blender to mash the pickled vegetables; Subsequently, stir the mashed mixture to make it evenly dispersed; Gradient dilute the mixture, and the diluent is sterile normal saline, and the dilution multiple is 10 -1 、10 -2 、10 -3 .
[0024] Preferably, step (3) is specifically as follows: Prepare MRS medium, perform autoclaving at 121 °C, and then inoculate the homogenate of the mixture diluted in step (2) onto the MRS medium. Put the medium into an anaerobic incubator and culture at 37 °C for 24 - 48 hours.
[0025] Preferably, step (4) includes: Regularly observe the colony growth on the medium, search for possible Lactobacillus paracasei colonies, which are round, smooth and milky white. Inoculate the suspected colonies onto a new MRS medium for purification culture to obtain pure colonies; Conduct biochemical identification on the purified strains among them, make a preliminary judgment according to the biochemical characteristics of the strains. Finally, routinely extract the DNA of the strains, perform 16S rRNA gene amplification and sequencing, and compare the sequencing results with the known Lactobacillus paracasei gene sequences to screen Lactobacillus paracasei.
[0026] Preferably, step (5) is specifically as follows: Store the Lactobacillus paracasei in a refrigerator at 4 °C for short - term storage; or, use a glycerol freezing tube and store it in a low - temperature refrigerator at - 80 °C for long - term storage.
[0027] Preferably, step (7) is expressed in parts by weight. Add 15 - 20 parts of granulated sugar to 100 parts of purified water; The temperature of the purified water is 70 °C;
[0028] Step (8) is expressed in parts by weight. 50 parts of purified water, 2 - 3 parts of crushed petals, 5 - 10 parts of sugar solution, and 50 parts of the remaining purified water; Heat up: to 65 - 70 °C; Mix and extract: Extract at 80 - 90 °C for 20 - 30 min.
[0029] Preferably, step (9) High - temperature sterilization: Continue to heat up to 90 - 100 °C and maintain this temperature for 20 - 30 min;
[0030] Step (10) Fermentation: After sterilization, cool the material to 35 - 37°C, inoculate with Lactobacillus paracasei, and ferment and culture at a constant temperature of 35 - 37°C for 48 - 72 h;
[0031] Step (11) Secondary fermentation: After sterilizing the material obtained from the fermentation and culture in step (10), cool the material to 35 - 37°C, inoculate with Lactobacillus paracasei, and ferment at a constant temperature for 48 - 72 h.
[0032] The present invention also provides a rose beverage prepared by any of the above - mentioned methods.
[0033] The present invention also provides the application of extracting Lactobacillus paracasei from Northeast sauerkraut to produce rose beverages.
[0034] As can be seen from the above - mentioned technical solutions, compared with the prior art, the present invention discloses a method for fermenting and making rose beverages using Lactobacillus paracasei, and the technical effects achieved are as follows:
[0035] By applying the Lactobacillus paracasei fermentation technology and carefully retaining the natural fragrance and comprehensive nutrition of rose petals, the rose components have certain health - care functions, such as antioxidant, beauty - enhancing, etc. Therefore, the beverage has potential health benefits. By optimizing the fermentation process and formula design, the beverage has a unique taste and flavor, while retaining the fragrance of roses and the sour - sweet taste of Lactobacillus paracasei, bringing a pleasant drinking experience to consumers. It accurately meets the urgent needs of consumers for healthy and natural foods.
[0036] Lactobacillus paracasei is an important lactic acid bacterium that can be used in the food fermentation process. It has health functions such as regulating intestinal flora and enhancing immunity. In the present invention, Lactobacillus paracasei (purity > 99% and activity, i.e., survival rate ≥ 90%) extracted and purified from Northeast sauerkraut can obtain a strain with excellent fermentation performance. It can be used to make fermented foods such as yogurt, pickles, and sauerkraut, which can not only improve the flavor and nutritional value of foods but also ensure the shelf life of foods.
[0037] Optimizing fermentation conditions:
[0038] 1. Control the extraction temperature (80 - 90°C) to retain the volatile aroma components of roses;
[0039] 2. Ferment in stages (initial pH 5.5 - 6.0, later pH 4.2 - 4.5) to promote the synthesis of β - phenylethyl alcohol and citronellol;
[0040] 3. Formula design: Adjust the mass ratio of rose petals to sugar solution, and the content of free polyphenols after fermentation increases by 30% - 40%.
[0041] The present invention can significantly increase the production of bacteriocin, thereby producing a food preservative with high anti-corrosion effect to replace traditional chemical preservatives and improve the safety and health of food.
[0042] The process method for extracting Lactobacillus paracasei from Northeast sauerkraut has wide uses and potential economic value, and can be used not only in food fermentation and preservation, the preparation of probiotic products, but also in many fields such as scientific research and application development. BRIEF DESCRIPTION OF THE DRAWINGS
[0043] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained according to the provided drawings without creative efforts.
[0044] Figure 1 It is the process flow chart provided for the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS
[0045] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the drawings in the embodiments of the present invention. Obviously, the described embodiments are only some of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.
[0046] The embodiment of the present invention discloses a method for fermenting and producing a rose beverage using Lactobacillus paracasei.
[0047] Raw materials not mentioned in the embodiments are all commercially available; equipment not mentioned is conventional production equipment; experimental methods not mentioned are conventional experimental methods, such as the processes of colony isolation and purification; except for being obtained by the extraction method, commercially available strains of Lactobacillus paracasei that meet the production requirements can also be used for the preparation of rose beverages, and the source is not limited.
[0048] Example 1
[0049] A method for fermenting and producing a rose beverage using Lactobacillus paracasei, comprising the following steps (see the process flow chart in Figure 1 ):
[0050] (1) Preparation of sauerkraut samples: Collect fresh sauerkraut from multiple farmers or different production batches in Northeast China. First, drain the water from the sauerkraut, and then use a clean knife to remove its inedible parts, such as the roots and withered leaves. To ensure purity, the knife will be rinsed with clean water before use. Then, cut the sauerkraut into small pieces about 1 cm square for subsequent operations;
[0051] (2) Sample homogenization: Put the cut small pieces of pickled Chinese cabbage into a blender and add enough water to cover the top of the pickled Chinese cabbage; After mixing and stirring for 10 - 20 min (15 min in this example), start the blender to mash the pickled Chinese cabbage; Subsequently, manually stir the mashed mixture to make it evenly dispersed; In order to isolate single colonies subsequently, perform gradient dilution on the mashed mixture, and the diluent is sterile normal saline, with a dilution factor of 10 -1 、10 -2 、10 -3 ;
[0052] (3) Inoculation and cultivation: Prepare MRS medium (designed for lactic acid bacteria cultivation, weigh each component according to the formula, mix peptone, trypticase, yeast extract, and agar, and salt solution can be routinely added, and adjust the pH value to between 5.7 - 6.0), and perform autoclaving at 121 °C for 15 min; Then, inoculate the homogenate of the mixture diluted in step (2) onto the MRS medium, and the inoculation method can choose the spread plate method or the pour plate method; After inoculation, put the medium into an anaerobic incubator and cultivate at 37 °C for 24 - 48 hours;
[0053] (4) Colony isolation and identification: During the cultivation process, regularly observe the colony growth on the medium and look for possible Lactobacillus paracasei colonies, which usually appear round, smooth, and milky white; Using aseptic operation techniques, inoculate the suspected colonies onto a new MRS medium for purification cultivation, and the purification method can choose the streak plate method, streak the colonies onto a new medium, and obtain pure colonies after cultivation; Next, perform biochemical identification on the purified strains among them (such as sugar fermentation tests and API test strips, etc.), and make a preliminary judgment based on the biochemical characteristics of the strains; Finally, (routinely) extract the DNA of the strains, perform 16S rRNA gene amplification and sequencing, and compare the sequencing results with the known Lactobacillus paracasei gene sequences to confirm the identity of the strains and screen Lactobacillus paracasei;
[0054] (5) Preservation of Lactobacillus paracasei: To preserve the identified Lactobacillus paracasei strains, there are two methods to choose from: One is to perform short-term preservation in a 4 °C refrigerator (this example);
[0055] The other is to use a glycerol freezing tube and perform long-term preservation in a -80 °C low-temperature refrigerator;
[0056] (6) Selection and treatment of rose petals: First, use a dustpan or sieve to remove impurities and select flawless rose petals; Subsequently, through multiple soakings and washings, ensure the purity and plumpness of the petals, and then perform crushing treatment to obtain crushed petals for subsequent use;
[0057] (7) Preparation of sugar solution: Slowly add granulated sugar to purified water at 70°C until it is completely dissolved to form a uniform sugar solution (in parts by weight, 100 parts of purified water and 15 - 20 parts of granulated sugar are added, 17 parts are added in this example);
[0058] (8) Mixed extraction: (in parts by weight) Add 50 parts of purified water, crushed petals (2 - 3 parts, 2.5 parts in this example) and sugar solution (5 - 10 parts, 7.5 parts in this example) to a fermentation tank. After heating to 65 - 70°C (67°C in this example), then add the remaining 50 parts of purified water and conduct mixed extraction (extract at 80 - 90°C for 20 - 30 min, 85°C for 25 min in this example) to obtain the material;
[0059] (9) High - temperature sterilization: Continue to heat up to 90 - 100°C (95°C in this example) and maintain this temperature for 20 - 30 min (25 min in this example) to ensure the sterile state of the material;
[0060] (10) Fermentation: After sterilization, cool the material to 35 - 37°C (36°C in this example), inoculate with Lactobacillus paracasei, and keep it at a constant temperature of 35 - 37°C (36°C in this example) for fermentation culture for 48 - 72 h (60 h in this example);
[0061] (11) Secondary fermentation: After sterilizing the material obtained from the fermentation culture in step (10), cool the material to 35 - 37°C (36°C in this example), inoculate with (purified) Lactobacillus paracasei, and ferment at a constant temperature for 48 - 72 h (60 h in this example) to promote the formation of flavor;
[0062] (12) Finished product: Sterilize the well - fermented rose beverage and fill it to obtain the rose beverage.
[0063] Combining Lactobacillus paracasei in Northeast sauerkraut with rose components creates a rose beverage with a unique flavor and taste, providing consumers with novel choices. Using microbial isolation and purification techniques, the dilution - coating plate method, selective medium screening, etc., combined with morphological observation, physiological and biochemical tests, and molecular biology identification, etc., to ensure that the extracted strain is Lactobacillus paracasei with high purity and high activity. By optimizing the fermentation process and formula design, Lactobacillus paracasei is fully integrated with rose components during the fermentation process. At the same time, the stability and taste of the beverage are improved. The above - mentioned example utilizes the fermentation characteristics of Lactobacillus paracasei, not only significantly enhancing the ability of rose petals as a fermentation substrate to produce β - phenylethyl alcohol and citronellol, but also greatly improving the efficiency of the entire fermentation process.
[0064] The various embodiments in this specification are described in a progressive manner. Each embodiment focuses on the differences from other embodiments. For the same or similar parts among the various embodiments, reference can be made to each other.
[0065] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be obvious to those skilled in the art. The general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but rather will be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A method for preparing a rose beverage by fermenting Lactobacillus paracasei, comprising the following steps: (1) Preparation of sauerkraut samples; (2) Sample homogenization: Add the chopped sauerkraut into clean water; After mixing and stirring, the mixture is crushed; then, the crushed mixture is stirred to make it uniformly dispersed; the mixture is gradiently diluted, and the diluent is sterile physiological saline; (3) Inoculation culture: the culture medium is sterilized by high pressure, and then the homogenate of the mixture diluted in step (2) is inoculated into the culture medium and cultured; (4) colony isolation and identification, screening of Lactobacillus paracasei; (5) Preservation of Lactobacillus paracasei; (6) Selection and processing of rose petals: First, remove impurities and select flawless rose petals, then soak and wash them multiple times, and then grind them to obtain grinded petals; (7) Preparation of sugar solution: Slowly add white sugar into purified water until it is completely dissolved to form a uniform sugar solution; (8) Mixed extraction: Add purified water, crushed petals and sugar solution into a fermentation tank, heat it, and then add the remaining purified water to mix and extract to obtain the material; (9) High temperature sterilization; (10) Fermentation: After sterilization, the material is cooled, Lactobacillus paracasei is added, and the fermentation culture is maintained at a constant temperature; (11) secondary fermentation; (12) Finished product: sterilize and fill to obtain rose drink.
2. The method according to claim 1, characterized in that Step (1) specifically comprises: collecting fresh Northeastern pickled cabbage, first draining the water from the pickled cabbage, then removing the inedible parts thereof, and cutting the pickled cabbage into small pieces.
3. The method according to claim 2, characterized in that Step (2) is specifically as follows: putting the chopped sauerkraut pieces into a masher and adding enough water to cover the top of the sauerkraut; after mixing and stirring for 10 to 20 minutes, starting the masher to mash the sauerkraut; then, stirring the mashed mixture to make it evenly dispersed; and diluting the mixture in a gradient manner, using sterile physiological saline as the dilution liquid, and the dilution multiple is 10 -1 , 10 -2 , 10 -3 .
4. The method according to claim 3, characterized in that Step (3) specifically comprises: preparing an MRS culture medium, sterilizing it under high pressure at 121° C., then inoculating the homogenate of the mixture diluted in step (2) onto the MRS culture medium, placing the culture medium in an anaerobic incubator, and culturing it at 37° C. for 24 to 48 hours.
5. The method according to claim 4, characterized in that Step (4) comprises: regularly observing the growth of the colonies on the culture medium, searching for possible Lactobacillus paracasei colonies that are round, smooth and milky white, inoculating the suspected colonies on a new MRS culture medium for purification and culture, and obtaining pure colonies; performing biochemical identification on the purified strains, making a preliminary judgment based on the biochemical characteristics of the strains, and finally, conventionally extracting DNA of the strains, performing 16S rRNA gene amplification and sequencing, and comparing the sequencing results with the known Lactobacillus paracasei gene sequences to screen the Lactobacillus paracasei.
6. The method according to claim 5, characterized in that Step (5) specifically comprises: storing the Lactobacillus paracasei in a refrigerator at 4° C. for a short term; or storing the Lactobacillus paracasei in a low-temperature refrigerator at -80° C. for a long term using a glycerol cryotube.
7. The method according to claim 5, characterized in that Step (7) is expressed in parts by weight: 15 to 20 parts of white sugar are added to 100 parts of purified water; the temperature of the purified water is 70° C. Step (8) is expressed in parts by weight as follows: 50 parts of purified water, 2-3 parts of crushed petals, 5-10 parts of sugar solution, and the remaining 50 parts of purified water; the heating is to 65-70°C; the mixed extraction is performed at 80-90°C for 20-30 minutes.
8. The method according to claim 7, characterized in that Step (9) high temperature sterilization: continue to raise the temperature to 90-100°C and maintain this temperature for 20-30 minutes; Step (10) fermentation: after sterilization, the material is cooled to 35-37° C., Lactobacillus paracasei is added, and fermentation is cultured at a constant temperature of 35-37° C. for 48-72 hours; Step (11) secondary fermentation: after sterilizing the material after fermentation and culture in step (10), the material is cooled to 35-37° C., Lactobacillus paracasei is added, and fermented at a constant temperature for 48-72 hours.
9. Rose beverage prepared by the method according to any one of claims 1 to 8.
10. The application of Lactobacillus paracasei extracted from Northeastern sauerkraut in making rose beverage.