CN116711774A - Preparation method of composite probiotic fermented milk with high interaction characteristics - Google Patents

Abstract

The invention discloses a preparation method of composite probiotic fermented milk with high interaction characteristics, and belongs to the technical field of probiotic fermented agents. The composite probiotic starter consists of a basic starter and an active probiotic starter, wherein the basic starter consists of streptococcus thermophilus S10 and lactobacillus delbrueckii subspecies bulgaricus ND02 according to the weight ratio of 1000:1, the active composite probiotics are prepared by mixing lactobacillus paracasei Zhang and bifidobacterium animalis subspecies Probio-M8 according to the proportion of 1:1, and the components are compounded in proportion. According to the preparation method of the composite probiotic fermented milk with high interaction characteristics, the basic starter Streptococcus thermophilus S10 and Lactobacillus delbrueckii subspecies bulgaricus ND02 in the composite probiotic starter can rapidly produce acid and viscosity and give the fermented milk a unique taste; the active probiotic starter culture of Lactobacillus paracasei Zhang and Lactobacillus bifidus subspecies Probio-M8 have excellent probiotic characteristics, and have various probiotic characteristics of regulating intestinal flora, immunity and the like.

Description

A method for preparing compound probiotic fermented milk with high interactive properties

Technical field

The present invention relates to the technical field of probiotic fermentation agents, and in particular to a method for preparing compound probiotic fermented milk with high interaction characteristics.

Background technique

With the continuous improvement of people's living standards, probiotics have received widespread attention due to their properties that can promote host health. Fermented dairy products serve as good carriers of probiotics, which can deliver probiotics into the intestinal tract of the host. Probiotics can not only provide more health value to fermented milk, but can also produce a variety of metabolites through fermentation to further enhance the flavor and taste of fermented milk. Probiotics need to be taken in a certain amount before they can exert their probiotic effects in the human body. At present, because the number of viable bacteria in fermented milk is generally low, many probiotics are unable to exert their full probiotic effect due to the low number of viable bacteria. There are different ecological relationships between different microorganisms, and different strains can achieve mutual symbiosis by utilizing metabolites from each other to make up for their own metabolic shortcomings. The interaction between probiotics and starter strains can not only increase the fermentation speed of the starter strains, but also increase the number of viable bacteria, making probiotic fermented dairy products have a higher number of viable bacteria. The present invention aims to obtain a preparation method of compound probiotic fermented milk with high interactive properties.

Contents of the invention

The object of the present invention is to provide a method for preparing compound probiotic fermented milk with high interactive characteristics. The basic starter cultures Streptococcus thermophilus S10 and Lactobacillus delbrueckii subsp. bulgaricus ND02 in the compound probiotic starter can quickly produce acid. , produce viscosity and give fermented milk a unique taste; the active probiotic starter culture Lactobacillus paracasei Zhang and Bifidobacterium animalis subspecies lactis Probio-M8 have excellent probiotic properties and have various probiotic properties such as regulating intestinal flora and immunity. .

In order to achieve the above object, the present invention provides a preparation method of compound probiotic fermented milk with high interactive characteristics, which includes the following steps:

S1. Preparation of raw milk: After the raw milk is accepted, impurities are removed to obtain fresh milk; the fresh milk is refrigerated in a 4°C thermostat, and the stirring device is used to continuously stir during the refrigeration period;

S2. Starter fermentation: homogenize the fresh milk with a high-pressure homogenizer after stirring to obtain a homogeneous emulsion, and sterilize the homogeneous emulsion to obtain a sterilized emulsion;

Then, a compound probiotic starter with high interactive characteristics is added to the sterilized emulsion, and the emulsion is fermented at 42°C to obtain a fermented fermented emulsion;

The compound probiotic starter culture consists of a basic starter culture and an active compound probiotic starter culture. The basic starter culture is composed of Streptococcus thermophilus S10 and Lactobacillus delbrueckii subsp. bulgaricus ND02 in a ratio of 1000:1. The probiotic starter culture is composed of Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 in a ratio of 1:1;

S3. Packaging and storage of fermented milk: Pack the fermented fermented milk into bags and refrigerate at 4°C after packaging.

Preferably, the total viable bacterial count of Streptococcus thermophilus S10, Lactobacillus delbrueckii subsp. bulgaricus ND02, Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 before fermentation is 1×10 ⁷ to 3× 10 ⁷ CFU/g.

Preferably, in step S1, after the raw milk is accepted, impurity removal is performed in a milk purifier, and the impurity level after impurity removal is ≤2ppm.

Preferably, in step S1, the stirring mode is side stirring, the stirring rate is 60 r/min, and the stirring time is 5 min.

Preferably, in step S2, the homogenization pressure of the high-pressure homogenizer is 12 to 18 MPa, and the homogenization time is 5 to 8 minutes.

Preferably, in step S2, the sterilization method is ultra-high temperature sterilization at 130-140°C for 2-3 seconds.

Preferably, in step S2, the fermentation time is 6 hours. After the fermentation, the total activity of Streptococcus thermophilus S10, Lactobacillus delbrueckii subsp. bulgaricus ND02, Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 The bacterial count is 1×10 ⁹ to 3×10 ⁹ CFU/g.

Preferably, in step S3, the fermented milk packaging process is to fill the fermented milk with inert gas under vacuum in a sterile environment, package the fermented milk into bags, and use ozone to sterilize the packaging environment for 1 hour before and after each start.

Therefore, the present invention adopts the above-mentioned preparation method of compound probiotic fermented milk with high interactive characteristics, and its technical effects are as follows:

(1) The basic starter cultures of the compound probiotic starter culture are Streptococcus thermophilus S10 and Lactobacillus delbrueckii subsp. bulgaricus ND02, which can quickly produce acid and viscosity and give fermented milk a unique taste; the active probiotic starter culture is Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 have excellent probiotic properties and have various probiotic properties such as regulating intestinal flora and immunity;

(2) Use the interaction method between basic starter culture and compound probiotics to increase the number of viable bacteria in fermented milk, which can maintain a high number of viable bacteria in fermented milk, thereby making the probiotics Fermented milk has a higher number of viable probiotic bacteria, which can better exert the probiotic effect of probiotics.

The technical solution of the present invention will be further described in detail below through the accompanying drawings and examples.

Description of the drawings

Figure 1 shows the difference analysis of volatile metabolites at the end of fermentation, where A is Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10-Lactobacillus paracasei Zhang and Lactobacillus delbrueckii subsp. bulgaricus ND02-thermophilus Differential metabolites of Streptococcus S10 non-volatile metabolites; B is Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10-Bifidobacterium animalis subsp. lactis Probio-M8 and Lactobacillus delbrueckii subsp. bulgaricus ND02- Differential metabolites of non-volatile metabolites of Streptococcus thermophilus S10; C is Lactobacillus delbrueckii subsp. bulgaricus ND02 - Streptococcus thermophilus S10 - Lactobacillus paracasei Zhang - Bifidobacterium animalis subsp. lactis Probio-M8 and De Differential metabolites of Lactobacillus bulgaricus subsp. bulgaricus ND02-Streptococcus thermophilus S10 non-volatile metabolites;

Figure 2 shows the metabolic pathways of non-volatile differential metabolite enrichment at the end of fermentation between Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 and Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10 respectively.

Detailed ways

The technical solution of the present invention will be further described below through the drawings and examples.

Unless otherwise defined, technical terms or scientific terms used in the present invention shall have the usual meaning understood by a person with ordinary skill in the field to which the present invention belongs.

It is obvious to those skilled in the art that the present invention is not limited to the details of the above-described exemplary embodiments, and that the present invention can be implemented in other specific forms without departing from the gist or essential characteristics of the present invention. Therefore, the embodiments should be regarded as illustrative and non-restrictive from any point of view, and the scope of the present invention is defined by the appended claims rather than the above description, and it is therefore intended that all claims falling within the claims All changes within the meaning and scope of equivalent elements are encompassed by the present invention, and any reference signs in a claim should not be construed as limiting the claim involved.

In addition, it should be understood that although this specification is described in terms of implementations, not each implementation only contains an independent technical solution. This description of the specification is only for the sake of clarity, and those skilled in the art should take the specification as a whole. , the technical solutions in each embodiment can also be appropriately combined to form other implementations that can be understood by those skilled in the art. These other embodiments are also covered by the scope of the present invention.

It should also be understood that the specific embodiments described above are only used to explain the present invention, and the protection scope of the present invention is not limited thereto. Any person familiar with the technical field can use the technology disclosed in the present invention within the scope of the present invention. Any equivalent substitution or change of the scheme and its inventive concept shall be covered by the protection scope of the present invention/invention.

Techniques, methods and devices known to those of ordinary skill in the relevant art may not be discussed in detail, but where appropriate, such techniques, methods and devices should be considered a part of the specification.

The disclosure contents of the prior art documents cited in the specification of the present invention are incorporated by reference into the present invention in their entirety, and are therefore part of the disclosure content of the present invention.

Embodiment 1

The invention discloses a compound probiotic starter with high interactive characteristics. The compound probiotic starter consists of a basic starter and an active compound probiotic starter. The basic starter consists of Streptococcus thermophilus S10 and Delbrueckii. Lactobacillus bulgaricus subsp. bulgaricus ND02 is compounded in a ratio of 1000:1. The active compound probiotic starter culture is composed of Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 in a ratio of 1:1.

The total viable bacterial count of Streptococcus thermophilus S10, Lactobacillus delbrueckii subsp. bulgaricus ND02, Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 before fermentation is 1×10 ⁷ to 3×10 ⁷ CFU. /g.

After fermentation, the total viable bacterial count of Streptococcus thermophilus S10, Lactobacillus delbrueckii subsp. bulgaricus ND02, Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 is between 1×10 ⁹ and 3×10 ⁹ CFU/g.

Fermented milk is prepared according to the preparation method of compound probiotic fermented milk with high interactive properties. The steps are as follows:

S1. Preparation of raw milk: After the raw milk is accepted, impurities are removed. The impurity removal process is carried out in the milk purifier. After the impurities are removed, the impurity level is ≤2ppm to obtain fresh milk; the fresh milk is refrigerated in a 4°C constant temperature box. During the refrigeration period Use the stirring device to continuously stir, the stirring method is side stirring, the stirring rate is 60r/min, and the stirring time is 5min;

S2. Starter fermentation: homogenize with a high-pressure homogenizer, the homogenization pressure is 12 to 18MPa, and the homogenization time is 5 to 8 minutes. After homogenization, a homogeneous emulsion is obtained, and then the homogeneous emulsion is sterilized (sterilization mode is 130 ~140℃ for 2~3 seconds ultra-high temperature sterilization) to obtain the sterilized emulsion;

Add a probiotic starter with high interactive properties to the sterilized emulsion, and ferment the emulsion for 6 hours at 42°C to obtain a fermented fermented emulsion;

S3. Packaging and storage of fermented milk: Pack the fermented fermented milk into bags and refrigerate at 4°C after packaging.

The number of viable probiotic bacteria in the fermented milk was checked, and the volatile metabolites and non-volatile metabolites of the fermented milk were measured.

Viable bacteria count detection method: Take the sample to be tested and place it at room temperature for 1 to 2 hours, and balance the temperature to be consistent with room temperature. Weigh 25g of milk powder sample and shake evenly in 225mL of physiological saline to obtain a mixed solution. Pipette 1 mL of the mixed solution into a 9 mL centrifuge tube, perform a 10-fold gradient dilution step by step, and select 0.2 mL of a suitable dilution solution for the pouring experiment. MRS agar medium was sterilized at 121°C for 15 minutes before use. After the pouring plate solidifies, it is placed in an anaerobic incubator at 37°C for 48 hours. Select a petri dish with a colony count between 30 and 300 for counting. The results of the total viable count of probiotics in fermented milk are shown in Table 1, where the experiment number is the number of parallels.

Table 1 Results of total viable bacteria count of probiotics in fermented milk

At the end of fermentation, using the basic starter Lactobacillus delbrueckii subsp. bulgaricus ND02 and Streptococcus thermophilus S10 fermentation system as reference, the addition of Lactobacillus paracasei Zhang, Bifidobacterium animalis subsp. lactis Probio-M8 and paracasei was analyzed respectively. Effect of Lactobacillus casei Zhang-Bifidobacterium animalis subsp. lactis Probio-M8 on volatile metabolites. At the end of fermentation, compared with the fermentation system of Lactobacillus delbrueckii subsp. bulgaricus ND02 and Streptococcus thermophilus S10, Lactobacillus paracasei Zhang, Bifidobacterium animalis subsp. lactis Probio-M8 and Lactobacillus paracasei Zhang-Animal Bi Fibrobacterium lactis subsp. Probio-M8 can increase the concentration of volatile metabolites, especially metabolites that contribute significantly to aroma components, such as diacetyl, acetoin, acetaldehyde, etc.

At the end of the fermentation, using the fermentation system of Lactobacillus delbrueckii subsp. bulgaricus ND02 and Streptococcus thermophilus S10 as a reference, the addition of Lactobacillus paracasei Zhang, Bifidobacterium animalis subsp. lactis Probio-M8 and Lactobacillus paracasei Zhang was analyzed. -The effect of Bifidobacterium animalis subsp. lactis Probio-M8 on non-volatile metabolites, the results are shown in Figure 1;

Figure 1A shows the differential metabolites of Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10-Lactobacillus paracasei Zhang and Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10 non-volatile metabolites;

Figure 1B shows the difference in non-volatile metabolites between Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10-Bifidobacterium animalis subsp. lactis Probio-M8 and Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10 Metabolites;

Figure 1C shows Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10-Lactobacillus paracasei Zhang-Bifidobacterium animalis subsp. lactis Probio-M8 and Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10 non- Differential metabolites of volatile metabolites.

At the end of fermentation, Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 had different effects on non-volatile metabolites, with 15 and 63 differential metabolites respectively; in fructose and mannose metabolism, galactose metabolism , lysine metabolism, antibiotic metabolism and other metabolic pathways have a greater impact.

Figure 2 shows the metabolic pathways of non-volatile differential metabolite enrichment at the end of fermentation between Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 and Lactobacillus delbrueckii subsp. bulgaricus ND02-Streptococcus thermophilus S10 respectively;

In summary, the probiotics Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 can be compounded with the basic starter Lactobacillus delbrueckii subsp. bulgaricus ND02 and Streptococcus thermophilus S10, which can increase the number of probiotics Viable bacterial counts and increased production of beneficial metabolites.

Therefore, the present invention adopts the above-mentioned preparation method of compound probiotic fermented milk with high interactive characteristics. The basic starter cultures of the compound probiotic starter, Streptococcus thermophilus S10 and Lactobacillus delbrueckii subsp. bulgaricus ND02, can quickly produce acid. , produce viscosity and give fermented milk a unique taste; the compound probiotics in the compound probiotic starter culture are Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8, which are probiotics with excellent probiotic properties and can regulate the intestinal tract. It has various probiotic properties such as intestinal flora and immunity.

Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. 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 solution of the present invention may be modified or equivalently substituted, but these modifications or equivalent substitutions cannot cause the modified technical solution to depart from the spirit and scope of the technical solution of the present invention.

Claims (8)

1. A method for preparing compound probiotic fermented milk with high interactive characteristics, which is characterized by comprising the following steps: S1. Preparation of raw milk: After the raw milk is accepted, impurities are removed to obtain fresh milk; the fresh milk is refrigerated in a 4°C thermostat, and the stirring device is used to continuously stir during the refrigeration period; S2. Starter fermentation: homogenize the fresh milk with a high-pressure homogenizer after stirring to obtain a homogeneous emulsion, and sterilize the homogeneous emulsion to obtain a sterilized emulsion; Then, a compound probiotic starter with high interactive characteristics is added to the sterilized emulsion, and the emulsion is fermented at 42°C to obtain a fermented fermented emulsion; The compound probiotic starter culture consists of a basic starter culture and an active compound probiotic starter culture. The basic starter culture is composed of Streptococcus thermophilus S10 and Lactobacillus delbrueckii subsp. bulgaricus ND02 in a ratio of 1000:1. The probiotic starter culture is composed of Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 in a ratio of 1:1; S3. Packaging and storage of fermented milk: Pack the fermented fermented milk into bags and refrigerate at 4°C after packaging. 2. A method for preparing compound probiotic fermented milk with high interaction characteristics according to claim 1, characterized in that Streptococcus thermophilus S10, Lactobacillus delbrueckii subsp. bulgaricus ND02, Lactobacillus paracasei Zhang The total viable bacterial count of Bifidobacterium animalis subsp. lactis Probio-M8 before fermentation is 1×10 ⁷ to 3×10 ⁷ CFU/g. 3. A method for preparing compound probiotic fermented milk with high interaction characteristics according to claim 1, characterized in that, in step S1, after the raw milk is accepted, the impurity removal is carried out in a milk purifier. Impurity level ≤2ppm. 4. A method for preparing compound probiotic fermented milk with high interactive characteristics according to claim 1, characterized in that in step S1, the stirring mode is side stirring, the stirring rate is 60r/min, and the stirring time is 5 minutes. 5. A method for preparing compound probiotic fermented milk with high interaction characteristics according to claim 1, characterized in that in step S2, the homogenization pressure of the high-pressure homogenizer is 12 to 18 MPa, and the homogenization time It is 5~8min. 6. The method for preparing compound probiotic fermented milk with high interaction characteristics according to claim 1, characterized in that in step S2, the sterilization method is ultra-high temperature sterilization at 130-140°C for 2-3 seconds. 7. A method for preparing compound probiotic fermented milk with high interaction characteristics according to claim 1, characterized in that in step S2, the fermentation time is 6 h. After the fermentation is completed, Streptococcus thermophilus S10, De The total viable bacterial count of Lactobacillus bulgaricus subsp. bulgaricus ND02, Lactobacillus paracasei Zhang and Bifidobacterium animalis subsp. lactis Probio-M8 is 1×10 ⁹ to 3×10 ⁹ CFU/g. 8. The preparation method of compound probiotic fermented milk with high interaction characteristics according to claim 1, characterized in that in step S3, the fermented milk packaging process is vacuum filling in a sterile environment. Inert gas is added, and the fermented milk is packed into bags. The packaging environment is sterilized with ozone for 1 hour before and after each start.