CN112167339A - Lactobacillus rhamnosus fermented milk and preparation method thereof - Google Patents
Lactobacillus rhamnosus fermented milk and preparation method thereof Download PDFInfo
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- CN112167339A CN112167339A CN202011107260.0A CN202011107260A CN112167339A CN 112167339 A CN112167339 A CN 112167339A CN 202011107260 A CN202011107260 A CN 202011107260A CN 112167339 A CN112167339 A CN 112167339A
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- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 abstract description 3
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 abstract description 3
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
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- Life Sciences & Earth Sciences (AREA)
- Microbiology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Dairy Products (AREA)
Abstract
The invention relates to the field of dairy products, in particular to lactobacillus rhamnosus fermented milk and a preparation method thereof. The invention provides a preparation method of lactobacillus rhamnosus fermented milk, which comprises the following steps: providing a lactobacillus rhamnosus proliferation agent, wherein the lactobacillus rhamnosus proliferation agent comprises a serous fluid of cooked red bean and pineapple pulp; mixing a lactobacillus rhamnosus proliferation agent and milk to provide a fermentation system; inoculating lactobacillus rhamnosus into the fermentation system, and performing fermentation treatment. Compared with other similar products, the rhamnose lactobacillus fermented milk prepared by the method has the advantages of natural edibility, good taste, low cost and the like, belongs to pure fermentation and no mixed bacteria pollution, and has long shelf life, thereby effectively solving the problems of other similar products in the prior art and having good industrialization prospect.
Description
Technical Field
The invention relates to the field of dairy products, in particular to lactobacillus rhamnosus fermented milk and a preparation method thereof.
Background
The lactobacillus rhamnosus is a dominant bacterium in intestinal tracts of healthy human bodies and has the beneficial effects of antagonizing pathogenic bacteria, regulating intestinal flora, synthesizing various vitamins, reducing cholesterol, preventing cancers, enhancing immunity and the like. At present, a large number of probiotics health care products containing lactobacillus rhamnosus are published at home and abroad, and are deeply welcomed by consumers. However, lactobacillus rhamnosus is generally difficult to grow and reproduce in milk, and although lactobacillus rhamnosus fermented milk is developed from different technical angles at home and abroad, some products have complex components and high cost, some production materials are not natural and edible, the taste is poor, some products are mixed fermentation and non-pure lactobacillus rhamnosus products, and some products have poor survivability and short shelf life and are difficult to market. Moreover, most of the existing lactobacillus rhamnosus fermented milk neglects the detection of mixed bacteria pollution, and the real content of lactobacillus rhamnosus is difficult to ensure.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention aims to provide a lactobacillus rhamnosus fermented milk and a preparation method thereof, which are used for solving the problems in the prior art.
In order to achieve the above and other related objects, the present invention provides, in one aspect, a method for preparing lactobacillus rhamnosus fermented milk, comprising:
1) providing a lactobacillus rhamnosus proliferation agent, wherein the lactobacillus rhamnosus proliferation agent comprises a serous fluid of cooked red bean and pineapple pulp;
2) mixing the lactobacillus rhamnosus proliferation agent provided in the step 1) with milk to provide a fermentation system;
3) inoculating the fermentation system provided by the step 2) with lactobacillus rhamnosus, and performing fermentation treatment.
In some embodiments of the present invention, the lactobacillus rhamnosus propagation agent comprises, based on 100 parts by weight of total weight:
2-20 parts of red beans;
5-30 parts of pineapple pulp.
In some embodiments of the present invention, the preparation method of the lactobacillus rhamnosus proliferation agent comprises: mixing the cooked red bean and pineapple pulp homogenate, preferably, further comprising: solid-liquid separation and/or sterilization.
In some embodiments of the invention, in the step 2), the content of the lactobacillus rhamnosus proliferation agent in the fermentation system is 3-30 wt%.
In some embodiments of the present invention, in the step 2), the milk is selected from one or more of raw milk, liquid milk and reconstituted milk.
In some embodiments of the invention, in step 2), the fermentation system further comprises an additive selected from a combination of one or more of a flavoring agent, an acid-base stabilizer, and a thickening agent.
In some embodiments of the invention, in the step 3), the Lactobacillus rhamnosus is Lactobacillus rhamnosus GG, Lactobacillus rhamnosus BD0016, Lactobacillus rhamnosus KF 7.
In some embodiments of the invention, in the step 3), the inoculation concentration of lactobacillus rhamnosus in the fermentation system is 0.1-200 × 106CFU/g。
In some embodiments of the invention, in the step 3), the fermentation temperature of the fermentation treatment is 36-38 ℃.
In some embodiments of the present invention, in the step 3), the fermentation time of the fermentation treatment is 10 to 20 hours.
In some embodiments of the invention, in step 3), the fermentation treatment is a stationary fermentation treatment.
The invention also provides lactobacillus rhamnosus fermented milk which is prepared by the preparation method of the lactobacillus rhamnosus fermented milk.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments, and other advantages and effects of the present invention will be apparent to those skilled in the art from the disclosure of the present specification.
The inventor discovers a lactobacillus rhamnosus proliferation agent through a large amount of practical researches, and further prepares lactobacillus rhamnosus fermented milk by the lactobacillus rhamnosus proliferation agent. The lactobacillus rhamnosus fermented milk prepared by the lactobacillus rhamnosus proliferation agent has the advantages of natural edibility, good taste, low cost, pure fermentation, no mixed bacteria pollution, long storage and survival time and the like, thereby greatly improving the product quality of the lactobacillus rhamnosus fermented milk and completing the invention on the basis.
The invention provides a preparation method of lactobacillus rhamnosus fermented milk, which comprises the following steps: 1) providing a lactobacillus rhamnosus proliferation agent, wherein the lactobacillus rhamnosus proliferation agent comprises a serous fluid of cooked red bean and pineapple pulp; 2) mixing the lactobacillus rhamnosus proliferation agent provided in the step 1) with milk to provide a fermentation system; 3) inoculating the fermentation system provided by the step 2) with lactobacillus rhamnosus, and performing fermentation treatment. In the preparation method provided by the application, the lactobacillus rhamnosus proliferation agent used can comprise pulp of cooked red beans and pineapple pulp, the lactobacillus rhamnosus proliferation agent is introduced into a fermentation system comprising milk and inoculated with lactobacillus rhamnosus, the growth of the lactobacillus rhamnosus in the fermentation system can be effectively promoted in the fermentation process, and in the prepared lactobacillus rhamnosus fermented milk, the pollution of mixed bacteria (such as aerobic pollution bacteria and the like, more specifically bacillus subtilis, escherichia coli, bacillus licheniformis and the like) can be effectively avoided.
The preparation method of the lactobacillus rhamnosus fermented milk provided by the invention can comprise the following steps: providing a lactobacillus rhamnosus proliferation agent, wherein the lactobacillus rhamnosus proliferation agent comprises a serous fluid of cooked red bean and pineapple pulp. Generally speaking, a lactobacillus rhamnosus proliferation agent needs to be prepared separately and completely, and then the prepared lactobacillus rhamnosus proliferation agent is mixed with milk to construct a fermentation system, so that the lactobacillus rhamnosus can be effectively promoted to grow in the fermentation system in the fermentation process.
In the above preparation method, red beans are generally mature seeds of Vigna angularis (Vigna angularis) which is an annual semi-wound herbaceous plant of the family Leguminosae, and are usually used by mixing with an appropriate amount of water, boiling to provide cooked red beans, and then mixing the cooked red beans with other raw material homogenate. Pineapple pulp generally refers to the pulp of the ripe fruit of pineapple (Ananas comosus (Linn.) Merr.) of the genus Ananadis of the family Anacardiaceae, which is usually fresh. The lactobacillus rhamnosus propagation agent comprises, by weight, 2-20 parts, 2-4 parts, 4-6 parts, 6-8 parts, 8-10 parts, 10-12 parts, 12-14 parts, 14-16 parts, 16-18 parts and 18-20 parts of red beans (by dry weight, namely the weight of uncooked red beans) by 100 parts of total weight. The lactobacillus rhamnosus propagation agent comprises, by weight, 5-30 parts, 5-10 parts, 10-15 parts, 15-20 parts, 20-25 parts, or 25-30 parts of pineapple pulp (by wet weight) based on 100 parts of total weight.
In the above preparation method, the Lactobacillus rhamnosus proliferation agent may comprise a pulp of cooked red bean and pineapple pulp. When the lactobacillus rhamnosus proliferation agent is prepared, generally, cooked red beans and pineapple pulp can be homogenates mixed to provide the lactobacillus rhamnosus proliferation agent. The lactobacillus rhamnosus propagation agent can also comprise water, and the content of the water can be proper usually. Generally, during the process of providing the cooked red bean and during the process of mixing the cooked red bean and the pineapple pulp slurry, an appropriate amount of water may be added to facilitate the preparation of the lactobacillus rhamnosus proliferation agent. For example, red beans may be mixed with an appropriate amount of water and cooked to provide cooked red beans. For another example, a pulp may be formed with the cooked red bean and pineapple pulp by an appropriate amount of water. For another example, the method may further include: solid-liquid separation and/or sterilization, whereby solids in the mixture resulting from the homogenizing and mixing can be removed to provide a homogeneous slurry that can be suitable for seed fermentation.
The preparation method of the lactobacillus rhamnosus fermented milk provided by the invention can also comprise the following steps: mixing the lactobacillus rhamnosus proliferation agent provided by the step 1) with milk to provide a fermentation system. The prepared lactobacillus rhamnosus proliferation agent can be mixed with ingredients such as milk and the like in a proper proportion to form a proper fermentation system for subsequent fermentation treatment.
In the preparation method, a proper amount of lactobacillus rhamnosus proliferation agent is usually required to be added into a fermentation system, and generally, a higher addition amount of the lactobacillus rhamnosus proliferation agent can effectively increase the number of viable lactobacillus rhamnosus, but when the addition amount is too high, the taste of the fermented milk is adversely affected, and the flavor is affected. For example, the amount of Lactobacillus rhamnosus propagation agent in the fermentation system may be 3 to 30 wt%, 3 to 4 wt%, 4 to 5 wt%, 5 to 6 wt%, 6 to 8 wt%, 8 to 10 wt%, 10 to 12 wt%, 12 to 15 wt%, 15 to 20 wt%, 20 to 25 wt%, or 25 to 30 wt%. The lactobacillus rhamnosus proliferation agent can be uniformly mixed in the fermentation system.
In the above-mentioned production method, it is usually necessary to contain a proper amount of milk in the fermentation system. The milk contained in the fermentation system can be raw milk or various dairy products which are suitable for fermenting the lactobacillus rhamnosus dairy products and are prepared from the raw milk. For example, applicable milk may be raw milk, liquid milk, reconstituted milk, or the like. The content of milk in the fermentation system can be adjusted appropriately for those skilled in the art, for example, the content of milk in the fermentation system can be more than or equal to 5 wt%, 5-10 wt%, 10-20 wt%, 20-30 wt%, 30-40 wt%, 40-50 wt%, 50-60 wt%, 60-70 wt%, 70-80 wt%, 80-90 wt%, or higher.
In the above preparation method, the fermentation system may further comprise an additive. The type and amount of suitable additives for the preparation of fermented milks should be adjustable for the person skilled in the art. For example, the additive may be selected from a combination of one or more of a flavoring agent (e.g., sucrose, etc.), an acid-base stabilizer (e.g., sodium citrate, etc.), a thickening agent (e.g., pectin, starch, etc.), and the like. For another example, the flavoring agent may be added in an amount of 2.5 to 7.5 wt%, 2.5 to 3.5 wt%, 3.5 to 4.5 wt%, 4.5 to 5.5 wt%, 5.5 to 6.5 wt%, or 6.5 to 7.5 wt% in the fermentation system. For another example, the amount of the acid-base stabilizer added to the fermentation system may be 0.1 to 1 wt%, 0.1 to 0.2 wt%, 0.2 to 0.4 wt%, 0.4 to 0.6 wt%, 0.6 to 0.8 wt%, or 0.8 to 1 wt%. For another example, the thickening agent may be added in an amount of 0.1 to 1 wt%, 0.1 to 0.2 wt%, 0.2 to 0.4 wt%, 0.4 to 0.6 wt%, 0.6 to 0.8 wt%, or 0.8 to 1 wt% in the fermentation system.
The preparation method of the lactobacillus rhamnosus fermented milk provided by the invention can also comprise the following steps: inoculating the fermentation system provided by the step 2) with lactobacillus rhamnosus, and performing fermentation treatment. After the lactobacillus rhamnosus proliferation agent is mixed with the milk and other ingredients in a proper proportion to form a proper fermentation system, the fermentation system can be subjected to fermentation treatment to provide the lactobacillus rhamnosus fermented milk.
In the above preparation method, Lactobacillus rhamnosus (Lactobacillus rhamnosus) is a kind of facultative anaerobic gram-positive bacteria belonging to Lactobacillus. In an embodiment of the present invention, Lactobacillus rhamnosus may be Lactobacillus rhamnosus GG (hansen ltd. in denmark), Lactobacillus rhamnosus BD0016 (bright milk industry ltd.), Lactobacillus rhamnosus KF7 (bright milk industry ltd.), etc.
In the preparation method, a proper amount of lactobacillus rhamnosus is generally required to be inoculated in the fermentation system, and the fermentation system or each component (such as milk, water and the like) of the fermentation system can be generally sterilized before the lactobacillus rhamnosus is inoculated, so as to ensure the pure fermentation effect in the fermentation treatment. The appropriate inoculation amount should be adjustable for the person skilled in the art, for example, the inoculation concentration of Lactobacillus rhamnosus in the fermentation system (based on the final concentration in the fermentation broth to be fermented) may be from 0.1 to 200X 106CFU/g、0.1-0.5×106CFU/g、0.5-1×106CFU/g、1-5×106CFU/g、5-10×106CFU/g、10-20×106CFU/g、20-40×106CFU/g、20-40×106CFU/g、40-60×106CFU/g、60-80×106CFU/g、80-100×106CFU/g、100-120×106CFU/g、120-140×106CFU/g、140-160×106CFU/g、160-180×106CFU/g, or 180-6CFU/g。
In the above production method, the fermentation treatment is usually a standing fermentation treatment. The conditions for the fermentation treatment may be suitably selected by those skilled in the art depending on the species of the bacterial species. For example, the fermentation temperature of the fermentation treatment may be 36 to 38 ℃, 36 to 36.5 ℃, 36.5 to 37 ℃, 37 to 37.5 ℃, or 37.5 to 38 ℃. For another example, the fermentation time of the fermentation treatment may be 10 to 20 hours, 10 to 12 hours, 12 to 14 hours, 14 to 16 hours, 16 to 18 hours, or 18 to 20 hours.
In a second aspect, the invention provides rhamnoseThe lactobacillus fermented milk is prepared by the preparation method of the lactobacillus rhamnosus fermented milk provided by the first aspect of the invention. The lactobacillus rhamnosus fermented milk prepared by the preparation method of the lactobacillus rhamnosus fermented milk basically has no pollution of mixed bacteria (such as aerobic pollution bacteria, and the like, and more specifically can be bacillus subtilis, escherichia coli, bacillus licheniformis, and the like) and has good bacterium increasing effect. For example, in an experiment in which fermented milk is subjected to aerobic contamination in a medium (e.g., LB medium), the total number of aerobic colonies may be < 1X 101CFU/g. As another example, in an experiment in which the fermented milk is subjected to a Lactobacillus rhamnosus count in a culture medium (e.g., TPY medium), the total number of colonies of Lactobacillus rhamnosus may be 3X 10 or more8CFU/g、≥4×108CFU/g、≥5×108CFU/g、≥6×108CFU/g、≥7×108CFU/g、≥8×108CFU/g, or more than or equal to 1X 109CFU/g. In addition, the lactobacillus rhamnosus fermented milk prepared by the preparation method of the lactobacillus rhamnosus fermented milk has longer storage period, the proportion of the number of viable bacteria can still be more than or equal to 20 percent, more than or equal to 25 percent or more than or equal to 30 percent after being refrigerated for one month (at 10 ℃), and the total number of the colonies of the lactobacillus rhamnosus can still be more than or equal to 1.0 multiplied by 10 in the lactobacillus rhamnosus counting experiment of the fermented milk in a culture medium (such as TPY culture medium)8CFU/g, or not less than 2X 108CFU/g。
Compared with other similar products, the rhamnose lactobacillus fermented milk prepared by the method has the advantages of natural edibility, good taste, low cost and the like, belongs to pure fermentation and no mixed bacteria pollution, and has long shelf life, thereby effectively solving the problems of other similar products in the prior art and having good industrialization prospect.
The invention of the present application is further illustrated by the following examples, which are not intended to limit the scope of the present application.
Example 1
Preparation of lactobacillus rhamnosus proliferation agent:
lactobacillus rhamnosus proliferation agent 1: soaking 1 part by weight of red beans in water for 6 hours, supplementing water to 9 parts by weight, cooking (100 ℃ for 1-2 hours), and cooling. Adding 2 parts by weight of fresh pineapple, adding 8 parts by weight of water, homogenizing and uniformly mixing. Keeping the temperature at 55 ℃ for 4h, filtering to remove residues, and sterilizing to obtain the lactobacillus rhamnosus proliferation agent.
Lactobacillus rhamnosus proliferation agent 2: soaking 1 part by weight of red beans in water for 10 hours, supplementing water to 9 parts by weight, cooking and cooling. Adding 4 parts by weight of fresh pineapple, adding 6 parts by weight of water, homogenizing and uniformly mixing. Preserving heat for 4h at 55 ℃, filtering and removing slag, and sterilizing to be used as a proliferation agent of the lactobacillus rhamnosus.
Lactobacillus rhamnosus proliferation agent 3: soaking 2 parts by weight of red beans in water for 16 hours, supplementing water to 8 parts by weight, cooking and cooling. Adding 5 parts by weight of fresh pineapple and 5 parts by weight of water, homogenizing and mixing uniformly. Preserving heat for 4h at 55 ℃, filtering and removing slag, and sterilizing to be used as a proliferation agent of the lactobacillus rhamnosus.
Blank control: soaking 1 part by weight of red beans in water for 16 hours, supplementing water to 9 parts by weight, cooking and cooling. Then 10 parts by weight of water was added thereto, and the mixture was homogenized and mixed. Keeping the temperature at 55 ℃ for 4h, filtering to remove residues, and sterilizing to obtain a blank control of a proliferation agent of the lactobacillus rhamnosus.
To determine the effect of proliferation agent of Lactobacillus rhamnosus, each prepared Lactobacillus rhamnosus proliferation agent is added into milk (fresh milk, produced by Bright milk Co., Ltd.) at a mass ratio of 10%, sterilized at 105 deg.C for 10min, and inoculated to a final concentration of about 1.0 × 106CFU/g (Lactobacillus rhamnosus KF7, Bright milk industries, Ltd.). Fermenting for 12h, sampling, diluting in gradient, counting aerobic contaminating bacteria with LB culture medium (after aerobic culture with LB agar culture medium at 37 ℃ for 48h, counting), counting lactobacillus rhamnosus with TPY culture medium (after anaerobic culture with TPY agar culture medium at 37 ℃ for 72h, counting), and the experimental results are shown in Table 1. As can be seen from the TPY viable bacteria counting results in the table 1, the prepared proliferation agent can promote the growth of lactobacillus rhamnosus in milk, while the proliferation agent added with pineapple has a better proliferation effect, the proliferation effect is further improved along with the increase of the addition amount of the pineapple, and the proliferation effect without the addition of the pineapple is relatively poor. As can be seen from the results of the total number of aerobic colonies in Table 1, in pure rhamnose milk rodsIn the process of bacteria fermentation, no mixed bacteria pollution is caused basically, and the bacteria increasing effect is reliable.
TABLE 1
| Total number of aerobic colonies (CFU/g) | TPY viable count (CFU/g) | |
| Production method 1 | <1×101 | 3.5×108 |
| Production method 2 | <1×101 | 7.2×108 |
| Production method 3 | <1×101 | 1.1×108 |
| Blank control | <1×101 | 1.8×106 |
Example 2
Preparation of lactobacillus rhamnosus fermented milk added with sodium citrate:
the lactobacillus rhamnosus fermented milk formula comprises the following components: 50 parts by weight of raw milk, 10 parts by weight of sucrose, 1 part by weight of sodium citrate, 0.2 part of a thickener (pectin (Zhengzhou Tianshun, food grade), the same applies below), the method in example 1 of lactobacillus rhamnosus proliferator 2 was followed to prepare the proliferator, the added amounts of the lactobacillus rhamnosus proliferator were: 5 parts by weight, 10 parts by weight, 15 parts by weight, 20 parts by weight, with the remainder made up to 100 parts by weight with the adjusted weight of water.
The preparation method comprises the following steps: preheating the mixed solution, heating, homogenizing at 65 deg.C and 20MPa, sterilizing at 105 deg.C for 10min, and cooling to 37 deg.C. Inoculation with Lactobacillus rhamnosus (the strain is as in example 1, final concentration is about 1.0X 106CFU/g), fermenting for 8 h. Sampling, gradient dilution, counting aerobic pollution bacteria by using LB culture medium, counting lactobacillus rhamnosus by using TPY culture medium, and the experimental result is shown in Table 2. In the sensory state evaluation, 5 professionals taste the fermentation product and give an evaluation result, if the evaluation result is not uniform, the evaluation result comprises two parts of acidity and mouthfeel according to the opinion of most people, wherein the acidity comprises: the taste is divided into four grades, namely sour and sweet, slightly sour, relatively sour and sour, and the taste comprises the following components: tasty, bitter and bitter. From the results of the viable cell count of TPY in table 2, it is found that the viable cell count of lactobacillus rhamnosus increases with the increase of the amount of the growth promoter added, and fermented milk is sour-sweet and tasty when the amount of addition is 10% or less, but the sensory state is biased to sour and bitter when the amount of addition is 15% or more, and the flavor is not good. As can be seen from the results of the total number of aerobic colonies in Table 2, there was almost no contamination by other bacteria during the fermentation of pure Lactobacillus rhamnosus, and the fermentation was pure.
TABLE 2
Example 3
Lactobacillus rhamnosus fermented milk storage viability:
lactobacillus rhamnosus fermented milk was prepared according to the formulation in example 2 (the amount of the proliferation agent added was 10%). After subpackaging, storing for 30 days at 10 ℃, diluting in a gradient way, counting aerobic pollution bacteria by using LB culture medium, counting lactobacillus rhamnosus by using TPY culture medium, and the experimental result is shown in table 3. From TPY in Table 3The result of bacteria counting shows that the number of the live bacteria in the sample is reduced little within 30 days of shelf life and can still reach 2.5 multiplied by 108CFU/g is far higher than the standard about the minimum limit of viable count of lactobacillus in the sanitary standard of lactobacillus beverage (GB 16321-: 1.0X 106The CFU/g "correlation provides that Lactobacillus rhamnosus in the sample has a high survival rate. As can be seen from the results of the total number of aerobic colonies in Table 3, the samples remained substantially free from contamination by infectious microbes during the storage period. In addition, the samples remained sour, sweet and palatable after storage.
TABLE 3
Example 4
Direct addition control test:
the preparation method comprises the following steps:
soaking 1 part by weight of red beans in water for 6 hours, supplementing water to 9 parts by weight, cooking and cooling. Adding 2 parts by weight of fresh pineapple and 8 parts by weight of water, homogenizing and mixing uniformly. Keeping the temperature at 50 ℃ for 3h, filtering and removing residues, and sterilizing to obtain the lactobacillus rhamnosus proliferation agent.
Preparing materials: 50 parts of raw milk, 10 parts of cane sugar, 1 part of sodium citrate, 0.1 part of thickening agent, 10 parts of lactobacillus rhamnosus proliferation agent, and the balance of water for adjusting the weight to be supplemented to 100 parts of weight. Preheating the mixed solution, heating, homogenizing at 65 deg.C and 20MPa, sterilizing at 105 deg.C for 10min, and cooling to 37 deg.C. Inoculation with Lactobacillus rhamnosus (the strain is as in example 1, final concentration is about 3.0X 106CFU/g), fermenting for 12 h. Sampling, diluting in gradient, counting aerobic pollution bacteria by using LB culture medium, and counting lactobacillus rhamnosus by using TPY culture medium.
The preparation method 2 comprises the following steps:
soaking 1 part by weight of red beans in water for 6 hours, supplementing water to 9 parts by weight, cooking, and cooling to prepare the red soybean milk. Preparing materials: 50 parts of raw milk, 10 parts of cane sugar, 1 part of sodium citrate, 0.1 part of thickening agent, 5 parts of red soybean milk and 1 part of fresh pineapple, and the rest is adjusted by water to make up the weight of the raw milk to 100 parts. The other operations are the same as those of the manufacturing method 1 of the embodiment 1.
The experimental result is shown in table 4, the preparation method 1 firstly prepares the lactobacillus rhamnosus proliferation agent separately and completely, and then mixes the prepared lactobacillus rhamnosus proliferation agent with milk to construct a fermentation system, so that the bacterium increasing effect is better; the contents of all substances in the formulas of the preparation method 2 and the preparation method 1 are the same, but the red beans and the pineapples are directly mixed with other components of the fermentation system in the preparation method 2, so that the bacterium increasing effect is poor.
TABLE 4
In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (10)
1. A preparation method of lactobacillus rhamnosus fermented milk comprises the following steps:
1) providing a lactobacillus rhamnosus proliferation agent, wherein the lactobacillus rhamnosus proliferation agent comprises a serous fluid of cooked red bean and pineapple pulp;
2) mixing the lactobacillus rhamnosus proliferation agent provided in the step 1) with milk to provide a fermentation system;
3) inoculating the fermentation system provided by the step 2) with lactobacillus rhamnosus, and performing fermentation treatment.
2. The method for preparing lactobacillus rhamnosus fermented milk according to claim 1, wherein in the step 1), the lactobacillus rhamnosus propagation agent comprises the following components in parts by weight based on 100 parts by weight:
2-20 parts of red beans;
5-30 parts of pineapple pulp.
3. The method for preparing lactobacillus rhamnosus fermented milk according to claim 1, wherein in the step 1), the method for preparing the lactobacillus rhamnosus proliferation agent comprises: mixing the cooked red bean and pineapple pulp homogenate, preferably, further comprising: solid-liquid separation and/or sterilization.
4. The method for preparing lactobacillus rhamnosus fermented milk according to claim 1, wherein in the step 2), the content of lactobacillus rhamnosus proliferation agent in the fermentation system is 3-30 wt%.
5. The method for preparing lactobacillus rhamnosus fermented milk according to claim 1, wherein in step 2), the milk is selected from one or more of raw milk, liquid milk and reconstituted milk.
6. The method for preparing lactobacillus rhamnosus fermented milk according to claim 1, wherein in step 2), the fermentation system further comprises an additive selected from one or more of a flavoring agent, an acid-base stabilizer, and a thickening agent.
7. The method for preparing Lactobacillus rhamnosus fermented milk according to claim 1, wherein in the step 3), the Lactobacillus rhamnosus is Lactobacillus rhamnosus GG, Lactobacillus rhamnosus BD0016, Lactobacillus rhamnosus KF 7.
8. The method for preparing lactobacillus rhamnosus fermented milk according to claim 1, wherein in the step 3), the inoculation concentration of lactobacillus rhamnosus in the fermentation system is 0.1-200 x 106CFU/g。
9. The method for preparing lactobacillus rhamnosus fermented milk according to claim 1, wherein in the step 3), the fermentation temperature of the fermentation treatment is 36-38 ℃;
and/or the fermentation time of the fermentation treatment is 10-20 h;
and/or the fermentation treatment is static fermentation treatment.
10. A lactobacillus rhamnosus fermented milk prepared by the method for preparing the lactobacillus rhamnosus fermented milk of any one of claims 1 to 9.
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