Method for preparing calendula petal sauce through fermentation of lactobacillus paracasei

Abstract

The invention discloses a method for preparing calendula petal sauce through fermentation of lactobacillus paracasei. Relates to the technical field of edible sauce processing. Comprising the following steps: preparing a pickled Chinese cabbage sample, homogenizing the sample, inoculating and culturing, separating and identifying bacterial colonies, preserving lactobacillus paracasei, harvesting fresh flower raw materials, pretreating fresh flowers, preparing a fermentation substrate, sterilizing, fermenting, adjusting the taste, and obtaining a finished product. The invention innovates the fermentation strain extraction source. The lactobacillus paracasei can better retain the natural fragrance and nutritional ingredients of the petals in the fermentation process, and meanwhile, a unique flavor can be possibly brought. Metabolite produced in the fermentation process of the lactic acid bacteria is beneficial to improving the nutritional value and health benefits of the petal sauce. The probiotic property of the lactobacillus paracasei is beneficial to promoting intestinal health, and the lactobacillus paracasei has excellent fermentation properties such as high acid yield, good salt tolerance and low-temperature growth performance, and can quickly reduce the pH value and inhibit the growth of other harmful microorganisms in the fermentation process.

Description

Technical Field

[0001] The invention relates to the technical field of edible sauce processing, and more particularly to a method for preparing marigold petal sauce by fermenting lactobacillus paracasei. Background Art

[0002] Calendula officinalis L., also known as marigold, marigold, etc., is an annual herbaceous plant of the genus Calendula in the Asteraceae family. Its flowers are orange or yellow, with a radial shape, and usually bloom in summer. Calendula has multiple edible functions, is rich in nutrients such as carotenoids, has a certain antioxidant effect, can help remove free radicals in the body, and delay aging. In addition, marigold also has certain antibacterial and anti-inflammatory properties, and can be used to make some natural food additives or condiments.

[0003] At present, most petal sauce products on the market are directly processed, lacking unique flavor and nutritional value. Due to various technical problems in processing, the taste may be monotonous and the aroma may be insufficient. At the same time, the nutrients in the petals, such as vitamins and antioxidants, may be damaged during processing, affecting their nutritional value. In addition, due to incomplete preservation measures or sterilization, the shelf life of some products may be short and easy to deteriorate. When the sanitary conditions during the production process are not up to standard. Some petal sauces may contain artificial additives such as pigments, flavors or preservatives, which may have an adverse effect on the health of consumers.

[0004] Therefore, being able to provide a method for preparing marigold petal paste by fermenting with Lactobacillus paracasei is a problem that those skilled in the art urgently need to solve. Summary of the invention

[0005] In view of this, the present invention provides a method for making marigold petal sauce by fermenting Lactobacillus paracasei. It is intended to provide a method for improving the taste and nutritional value of petal sauce by fermenting Lactobacillus paracasei (extracted from sauerkraut). Explore a method that can retain the fragrance of marigold flowers and is convenient to eat. Through the fermentation technology of Lactobacillus paracasei, the quality and taste of petal sauce are significantly improved, while the natural fragrance and rich nutrients of petals are retained to meet the needs of consumers for health and natural food. Not only can the shelf life of petal sauce be extended to ensure food safety, but also the risk of microbial contamination can be reduced by strict sanitation control and sterilization. In addition, the present invention increases the added value of the product by improving the nutritional value and market competitiveness of petal sauce, helps to promote the development of local characteristic industries and promote the innovation of food processing technology. By rationally utilizing petal resources, the present invention also conforms to the concept of environmental protection and sustainable development, and provides low-sugar, low-additive petal sauce selection for consumers pursuing a healthy lifestyle, thereby enriching the market product variety and meeting specific consumer needs.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] A method for preparing marigold petal sauce by fermenting Lactobacillus paracasei comprises the following steps:

[0008] (1) Preparation of sauerkraut samples:

[0009] (2) Sample homogenization: Add the chopped sauerkraut pieces to clean water, mix and stir, mash, and then stir the mashed mixture to make it evenly dispersed; dilute the mixture in a gradient manner using sterile saline;

[0010] (3) Inoculation culture: prepare a culture medium, sterilize it under high pressure, and then inoculate the homogenate of the mixture diluted in step (2) onto the culture medium and culture it;

[0011] (4) Colony isolation and identification: screening of Lactobacillus paracasei;

[0012] (5) Storage of Lactobacillus paracasei:

[0013] (6) Harvesting fresh flower raw materials: Selecting pest-free, plump and uniform-sized marigolds. When making petal sauce, pick the marigolds when they are half-opened for later use;

[0014] (7) Flower pretreatment: remove debris, remove unripe and insect-infested marigolds, and then soak the selected marigolds in water to allow them to absorb water. Repeat this process several times until the marigolds are clean and crushed for later use.

[0015] (8) Preparing a fermentation medium: adding an appropriate amount of sterile purified water to the crushed petals, and then adding glucose to obtain a fermentation medium;

[0016] (9) Sterilization: sterilize the fermentation substrate in step (8) by conventional pasteurization;

[0017] (10) fermentation: after the sterilized fermentation substrate is cooled, Lactobacillus paracasei is inoculated and fermented in a sterile environment to obtain a fermentation material;

[0018] (11) Adjusting the taste: adding appropriate amounts of white sugar, edible salt and citric acid to the fermented material in step (10) to obtain a mixed material;

[0019] (12) Finished product: The mixed material obtained in step (11) is sterilized by conventional pasteurization method and then cooled to obtain marigold petal paste.

[0020] Preferably, step (1) specifically comprises: collecting fresh sauerkraut from the Northeast region, draining the sauerkraut, removing inedible parts, and cutting the sauerkraut into small pieces.

[0021] Preferably, step (2) is specifically as follows: pouring the chopped sauerkraut pieces into a masher, adding clean water, the amount of clean water should cover the top of the sauerkraut, mixing and stirring for 10 to 20 minutes, turning on the masher, mashing the sauerkraut, and then stirring the mashed mixture to make it evenly dispersed; gradient diluting the mashed mixture, using sterile saline, and setting the dilution multiple to 10 -1 , 10 -2 , 10 -3 .

[0022] Preferably, step (3) specifically comprises: preparing an MRS culture medium, sterilizing it under high pressure at 121° C. for 15 min, then inoculating the homogenate of the mixture diluted in step (2) onto the MRS culture medium, placing the inoculated culture medium in an anaerobic incubator, and culturing it at 37° C. for 24 to 48 hours.

[0023] Preferably, step (4) comprises: regularly observing the growth on the culture medium, identifying possible Lactobacillus paracasei colonies, which are round, smooth and milky white, inoculating suspected colonies on new MRS culture medium, purifying and culturing to obtain pure colonies; performing biochemical identification on the purified strains, observing the fermentation of different sugars by the strains, performing preliminary identification based on the biochemical characteristics of the strains, conventionally extracting DNA of the strains, performing 16S rRNA gene amplification and sequencing, comparing the sequencing results with the known Lactobacillus paracasei gene sequences, and screening Lactobacillus paracasei.

[0024] Preferably, step (5) specifically comprises: storing the identified Lactobacillus paracasei strain in a refrigerator at 4°C for a short term, or storing it in a low-temperature refrigerator at -80°C using a glycerol cryovial for a long term.

[0025] Preferably: the half-open state in step (6): the degree of opening is 50% to 60%;

[0026] Step (7) soaking time: 30 to 40 minutes;

[0027] The amount of glucose added in step (8) is 1% by weight.

[0028] Step (10) specifically comprises: cooling the sterilized fermentation substrate to 35-37° C., inoculating Lactobacillus paracasei at a 1% inoculum amount, and fermenting in a sterile environment at 37° C. for 5-7 days to obtain a fermentation material;

[0029] In step (11), the mass ratio of the fermentation material, white sugar, edible salt and citric acid is 100:10-15; 1.5-2.5:0.1-0.3.

[0030] The beneficial effects are:

[0031] Granulated Sugar: Provides sweetness to balance the sour taste of fermentation and acts as a natural preservative to enhance shelf life.

[0032] Salt:

[0033] Inhibit the growth of bacteria, assist in preservation, and enhance flavor levels.

[0034] Lower than traditional kimchi (2% to 5%), to avoid covering up the fragrance of marigolds.

[0035] Citric Acid:

[0036] Adjust the pH value and stabilize the flavor of the fermentation product; add a small amount of acidity but do not affect the activity of the bacteria.

[0037] Furthermore, it can be fine-tuned according to the target taste.

[0038] The order of addition is to first dissolve sugar, salt and citric acid in a small amount of sterile water, and then mix them evenly with the fermentation material to avoid excessive local concentration.

[0039] The present invention also provides marigold petal jam prepared by any of the above methods.

[0040] The present invention also provides the use of extracting Lactobacillus paracasei from northeastern pickled cabbage to prepare marigold petal sauce.

[0041] It can be seen from the above technical scheme that, compared with the prior art, the present invention discloses a method for preparing marigold petal jam by fermenting Lactobacillus paracasei, and the technical effects achieved are:

[0042] The invention solves the technical problems existing in the production process of existing petal sauce. First, the traditional petal sauce may not fully retain the natural aroma and nutrients of the petals during the fermentation process, which is mainly due to the limitations of commonly used lactic acid bacteria in terms of fermentation efficiency and flavor formation. Secondly, the petal sauce products on the market often lack characteristics and are difficult to meet consumers' pursuit of novel and unique flavors. In addition, the traditional petal sauce may be contaminated by unexpected microorganisms during the production process, affecting product quality and safety. By using Lactobacillus paracasei extracted from Northeast sauerkraut for fermentation, these technical difficulties are effectively overcome.

[0043] The source of fermentation strain extraction has been innovated. As a non-traditional lactic acid bacteria, Lactobacillus paracasei can better retain the natural aroma and nutrients of petals during the fermentation process, and may also bring a unique flavor, enhancing the flavor and taste of petal sauce. The metabolites produced by this lactic acid bacteria during the fermentation process help to improve the nutritional value and health benefits of petal sauce. In addition, the probiotic properties of Lactobacillus paracasei help promote intestinal health, providing consumers with a food option with potential health benefits. Through this innovative fermentation method, the prepared petal sauce product has higher market competitiveness, meets consumers' demand for healthy and natural food, and also promotes the application innovation of Lactobacillus paracasei in the field of food processing. Extracting Lactobacillus paracasei from Northeastern sauerkraut has rich strain resources and excellent fermentation performance. The unique microbial community structure formed by Northeastern sauerkraut during the fermentation process provides a rich strain resource for screening Lactobacillus paracasei with excellent characteristics. Lactobacillus occupies a major position in the bacterial flora of Northeastern sauerkraut, including Lactobacillus paracasei. This diversity makes the Lactobacillus paracasei extracted from Northeastern pickled cabbage have unique regional characteristics, which may have adapted to local environmental factors such as climate and soil, and show different fermentation characteristics and metabolites from other regions. In addition, Lactobacillus paracasei has excellent fermentation properties such as high acid production, good salt tolerance and low-temperature growth performance. During the fermentation process, it can quickly reduce the pH value and inhibit the growth of other harmful microorganisms. At the same time, it can still maintain a certain growth and fermentation ability in a high-salt and low-temperature environment. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0045] Figure 1 The process flow chart provided by the present invention. DETAILED DESCRIPTION

[0046] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0047] The embodiment of the invention discloses a method for preparing marigold petal jam by fermenting Lactobacillus paracasei.

[0048] The raw materials not mentioned in the examples are all commercially available; the equipment not mentioned is conventional production equipment; the experimental methods not mentioned are conventional experimental methods, such as colony isolation and purification processes; in addition to obtaining Lactobacillus paracasei by extraction method, commercially available strains that meet production requirements can also be used to prepare marigold petal jam, without limitation on the source.

[0049] Example 1

[0050] A method for preparing marigold petal sauce by fermenting with Lactobacillus paracasei comprises the following steps (process flow see Figure 1 ):

[0051] (1) Preparation of sauerkraut samples: fresh sauerkraut samples were collected from the Northeast region. To ensure the representativeness of the samples, it was best to collect them from multiple households or different batches. The sauerkraut samples were placed in a dry place and inedible parts such as roots and yellow leaves were removed using a knife. The knife needed to be rinsed with clean water before use to avoid contamination. The sauerkraut was then cut into small pieces, about 1 cm square, for subsequent processing.

[0052] (2) Sample homogenization: Pour the chopped sauerkraut into a blender and add clean water. The amount of clean water should cover the top of the sauerkraut. Mix and stir for 10 to 20 minutes. Turn on the blender to blend the sauerkraut sample. Then manually stir the blended mixture to make it evenly dispersed. Dilute the blended mixture in a gradient manner. Sterile saline is usually used. The dilution multiple can be set to 10. -1 , 10 -2 , 10 -3 etc., to facilitate the subsequent isolation of single colonies;

[0053] (3) Inoculation culture: prepare MRS (De Man, Rogosa and Sharpe) medium (a medium for culturing lactic acid bacteria, weigh each component according to the formula, mix peptone, trypsin casein, yeast extract, and agar, and conventionally add salt solution to adjust the pH to 5.7-6.0), and then sterilize by high pressure at 121° C. for 15 min; then, inoculate the homogenate of the diluted mixture onto the MRS medium, and a certain amount of the dilution can be evenly coated or poured onto the medium by a plate coating method or a pouring plate method; place the inoculated medium into an anaerobic incubator and culture at 37° C. for 24-48 hours (36 hours in this embodiment);

[0054] (4) Colony isolation and identification: During the culture process, regularly observe the growth on the culture medium to identify possible Lactobacillus paracasei colonies, which are usually round, smooth, and milky white; use aseptic operation techniques to inoculate suspected colonies onto new MRS culture medium for purification culture. The plate streak method can be used to streak the colonies onto new culture medium to obtain pure colonies after culture; perform biochemical identification (such as sugar fermentation test) on the purified strains, observe the fermentation of different sugars by the strains (such as API test strips), perform preliminary identification based on the biochemical characteristics of the strains, extract the DNA of the strains, perform 16S rRNA gene amplification and sequencing, compare the sequencing results with the known Lactobacillus paracasei gene sequences, and screen for Lactobacillus paracasei;

[0055] (5) Storage of Lactobacillus paracasei: The identified Lactobacillus paracasei strain was stored in a refrigerator at 4° C. for a short term (in this embodiment), or stored in a low-temperature refrigerator at -80° C. using a glycerol cryovial for a long term;

[0056] (6) Harvesting fresh flower raw materials: Selecting pest-free, plump and uniform-sized marigolds. When making petal jam, the degree of opening of the marigolds has a great influence on the taste and aroma of the final product. When the marigolds are half-opened, the aroma is the most intense, which is an ideal choice for making petal jam. At this time, the petals have begun to unfold but have not yet fully opened, and the aroma is well retained in the petals. Therefore, the petal raw materials of the present invention select fresh marigolds with an opening degree of 50% to 60%, which are picked for later use;

[0057] (7) Flower pretreatment: Use a dustpan or sieve to remove debris such as leaves and branches, then remove the unfull and insect-infested marigolds, and then soak the selected marigolds in water for 30 to 40 minutes (35 minutes in this embodiment) to allow the marigolds to absorb water. Repeat several times until the Sophora japonica flowers are cleaned and crushed for later use;

[0058] (8) Preparation of fermentation matrix: adding an appropriate amount of sterile purified water to the crushed petals to provide the moisture required for the growth of microorganisms; then adding 1% glucose as a carbon source to provide energy and substances required for the synthesis of new cells for the microorganisms to obtain the fermentation matrix;

[0059] (9) Sterilization: sterilize the fermentation substrate in step (8) by conventional pasteurization;

[0060] (10) Fermentation: After the sterilized fermentation substrate is cooled to 35-37° C., Lactobacillus paracasei is inoculated at a 1% inoculum amount and fermented in a sterile environment at 37° C. for 5-7 days (6 days in this embodiment) to obtain a fermentation material;

[0061] (11) Adjusting the taste: adding appropriate amounts of white sugar, edible salt and citric acid (mass ratio is 100:10-15:1.5-2.5:0.1-0.3, and in this embodiment, 100:12:2:0.2) to the fermented material in step (10) to obtain a mixed material;

[0062] (12) Finished product: The mixed material obtained in step (11) is sterilized by conventional pasteurization method, and then cooled (20-25° C.) to obtain marigold petal paste.

[0063] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0064] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may 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 the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing marigold petal sauce by fermenting Lactobacillus paracasei, comprising the following steps: (1) Preparation of sauerkraut samples: (2) Sample homogenization: Add the chopped sauerkraut pieces to clean water, mix and stir, mash, and then stir the mashed mixture to make it evenly dispersed; dilute the mixture in a gradient manner using sterile saline; (3) Inoculation culture: prepare a culture medium, sterilize it under high pressure, and then inoculate the homogenate of the mixture diluted in step (2) onto the culture medium and culture it; (4) Colony isolation and identification: screening of Lactobacillus paracasei; (5) Storage of Lactobacillus paracasei: (6) Harvesting fresh flower raw materials: Selecting pest-free, plump and uniform-sized marigolds. When making petal sauce, pick the marigolds when they are half-opened for later use; (7) Flower pretreatment: remove debris, remove unripe and insect-infested marigolds, and then soak the selected marigolds in water to allow them to absorb water. Repeat this process several times until the marigolds are clean and crushed for later use. (8) Preparing a fermentation medium: adding an appropriate amount of sterile purified water to the crushed petals, and then adding glucose to obtain a fermentation medium; (9) Sterilization: sterilize the fermentation substrate in step (8) by conventional pasteurization; (10) fermentation: after the sterilized fermentation substrate is cooled, Lactobacillus paracasei is inoculated and fermented in a sterile environment to obtain a fermentation material; (11) Adjusting the taste: adding appropriate amounts of white sugar, edible salt and citric acid to the fermented material in step (10) to obtain a mixed material; (12) Finished product: The mixed material obtained in step (11) is sterilized by conventional pasteurization method and then cooled to obtain marigold petal paste.

2. The method according to claim 1, characterized in that Step (1) specifically comprises: collecting fresh sauerkraut from the northeast region, draining the sauerkraut, removing inedible parts, and cutting the sauerkraut into small pieces.

3. The method according to claim 2, characterized in that Step (2) is specifically as follows: pour the chopped sauerkraut pieces into a masher, add clean water to cover the top of the sauerkraut, mix and stir for 10 to 20 minutes, turn on the masher, mash the sauerkraut, and then stir the mashed mixture to make it evenly dispersed; dilute the mashed mixture in a gradient manner, use sterile saline, and set the dilution multiple to 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. for 15 min, then inoculating the homogenate of the mixture diluted in step (2) onto the MRS culture medium, placing the inoculated 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 on the culture medium, identifying possible Lactobacillus paracasei colonies, which are round, smooth and milky white, inoculating the suspected colonies on a new MRS culture medium, purifying and culturing them, and obtaining pure colonies; performing biochemical identification on the purified strains, observing the fermentation of different sugars by the strains, performing preliminary identification based on the biochemical characteristics of the strains, conventionally extracting DNA of the strains, performing 16SrRNA gene amplification and sequencing, comparing the sequencing results with the known Lactobacillus paracasei gene sequences, and screening Lactobacillus paracasei.

6. The method according to claim 5, characterized in that Step (5) specifically includes: storing the identified Lactobacillus paracasei strain in a refrigerator at 4° C. for a short term, or storing it in a low-temperature refrigerator at -80° C. using a glycerol cryovial for a long term.

7. The method according to claim 6, characterized in that The half-open state in step (6): the degree of opening is between 50% and 60%; Step (7) soaking time: 30 to 40 minutes; The amount of glucose added in step (8) is 1% by weight.

8. The method according to claim 7, characterized in that Step (10) specifically comprises: cooling the sterilized fermentation substrate to 35-37° C., inoculating Lactobacillus paracasei at a 1% inoculum amount, and fermenting in a sterile environment at 37° C. for 5-7 days to obtain a fermentation material; In step (11), the mass ratio of the fermentation material, white sugar, edible salt and citric acid is 100:10-15:1.5-2.5:0.1-0.

3.

9. Marigold petal jam prepared by the method according to any one of claims 1 to 8.

10. Application of Lactobacillus paracasei extracted from Northeastern sauerkraut in making marigold petal sauce.

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