CN107996705A - A kind of acidified milk rich in bifidobacterium longum and preparation method thereof - Google Patents

Abstract

The invention discloses a method for preparing fermented milk rich in Bifidobacterium longum. The preparation method comprises the following steps: (1) respectively inoculating Bifidobacterium longum and Leuconostoc enterococci BD01710 into raw milk, stirring and mixing evenly , at a temperature of 30-37°C, leave it to ferment until the titrated acidity is 70-95°T to obtain fermented milk; (2) cool the fermented milk prepared in step (1) to 16-22°C to obtain cooled fermented milk (3) Filling the cooled fermented milk into a packaging container, then transferring it to an environment with a temperature of 2-10° C. for refrigeration and then ripening for 12-24 hours to obtain fermented milk rich in Bifidobacterium longum. The fermented milk rich in Lactobacillus plantarum prepared by the invention is rich in nutrition, good in taste, simple and easy to prepare, and has a high content of active Bifidobacterium longum in the fermented milk, which can effectively regulate the balance and overall function of intestinal flora after eating. Intestinal action and other probiotic functions have great market application potential and economic value.

Description

Fermented milk rich in Bifidobacterium longum and preparation method thereof

technical field

The invention relates to the technical field of microbial fermentation, in particular to fermented milk rich in Bifidobacterium longum and a preparation method thereof.

Background technique

The Streptococcus thermophilus-Lactobacillus bulgaricus combination is currently the most classic starter combination in fermented milk. However, these two lactic acid bacteria have limited tolerance to the digestive tract environment and are not ideal probiotics. Therefore, the industry has been trying to develop a fermented milk system based on probiotics.

Probiotics are a class of active microorganisms that are beneficial to the host. They are a general term for active beneficial microorganisms that colonize the human intestinal tract and reproductive system and can produce definite health effects to improve the host's micro-ecological balance and exert beneficial effects. The probiotics commonly used in the human body and long body mainly include: Clostridium butyricum, lactic acid bacteria, bifidobacteria, and Lactobacillus acidophilus. Bifidobacteria are a type of lactic acid bacteria that play an important role in the health of the intestinal tract and the body. Studies have shown that Bifidobacterium, as a physiological beneficial bacterium, has many important physiological functions for human health, such as biological barrier, nutrition, anti-tumor effect, immune enhancement, improvement of gastrointestinal function, and anti-aging. In middle-aged and old age, the number of bifidobacteria begins to decline, and at the same time harmful bacteria such as Clostridium proliferate, which will adversely affect gastrointestinal health. In addition, diet, stress, drug use, and environmental changes can also lead to an imbalance in the gut flora. Therefore, supplementing bifidobacteria through food has important nutritional health and disease intervention effects on the human body.

Bifidobacterium longum (Bifidobacterium longum, referred to as BL bacteria) is often called dragon root bacteria, which is a kind of probiotics. Bifidobacterium longum is generally used in food, and has the effect of regulating intestinal health. Conforms to the bifidobacteria approved by the US FDA and can be used in general food. According to the data verified by medical research institutes, Bifidobacterium longum has the effect of regulating the human intestinal tract and improving health. Therefore, Bifidobacterium longum is a research hotspot in the field of food. The fermented milk based on Bifidobacterium longum currently on the market adds Bifidobacterium longum powder after fermentation, ignoring the effect of BL bacteria metabolites on the body's health. The difficulty in the growth of Bifidobacterium longum in milk has become a bottleneck limiting its large-scale application in the food field.

Contents of the invention

The purpose of the present invention is to solve the problem that Bifidobacterium longum is difficult to grow in milk. Through experiments, it is found that Leuconostoc mesenteroides (Leuconostoc mesenteroides) BD01710 has a growth-promoting effect on Bifidobacterium longum. Fermented milk containing bifidobacterium longum and a preparation method of the fermented milk.

In order to achieve the above object, the present invention is specifically achieved through the following technical solutions:

A method for preparing fermented milk rich in Bifidobacterium longum, the preparation method comprising the following steps: (1) inoculating Bifidobacterium longum (Bifidobacterium longum) and Leuconostocmesenteroides (Leuconostocmesenteroides) BD01710 into raw milk respectively, Stir and mix evenly, and leave to ferment at a temperature of 30-37°C until the titrated acidity is 70-95°T to obtain fermented milk; (2) cooling the fermented milk prepared in step (1) to 16-22°C to obtain Cooling the fermented milk; (3) filling the cooled fermented milk into a packaging container, and then transferring it to an environment with a temperature of 2-10°C for refrigerating and then ripening for 12-24 hours to obtain fermented milk rich in Bifidobacterium longum .

Further, the Leuconostoc mesenteroides described in step (1) is Leuconostoc mesenteroides BD01710 with the preservation number CGMCC NO.6432.

Further, in step (1), the inoculum amount of the Bifidobacterium longum is 10 ⁶ -10 ⁷ CFU/mL, and the inoculum amount of Leuconostoc enterolis BD01710 is 10 ⁶ -10 ⁷ CFU/mL.

Further, in step (1), the ratio of the inoculation amount of Bifidobacterium longum to Leuconostoc mesenteroides is 1:10-10:1.

Further, in step (2), the cooling method is the method of stirring after passing through the ice bath or the method of turning over the plates.

Further, in step (1), the raw milk is 5-15% whole milk powder or skim milk powder, 0-8% sucrose and water supplemented to 100%, after dissolving, mixing and homogenizing , sterilized, and cooled; the percentages are mass percentages.

Further, in step (1), the raw milk is prepared by adding 3-8% sucrose and 0-1.5ppm manganese gluconate after standardizing the raw milk, dissolving, mixing, homogenizing, sterilizing, and cooling; The percentages are mass percentages.

The present invention also provides fermented milk rich in Bifidobacterium longum prepared by the above-mentioned preparation method.

The positive progress effect of the present invention is:

1. For the fermented milk rich in Bifidobacterium longum provided by the present invention, the fermentation medium used in the preparation process has a wide range of sources, low cost, and is natural and safe; through co-cultivation experiments, it was found that Leuconostoc enterococci BD01710 has a strong effect on Bifidobacterium longum growth-promoting effect, and on this basis, by preparing the fermented milk rich in Bifidobacterium longum of the present invention, a binary fermentation system based on Leuconostoc enterococci and Bifidobacterium longum was constructed in the fermented product. The system can realize the growth and curd of Bifidobacterium longum in the milk medium, and the number of viable Bifidobacterium longum can reach more than 5×10 ⁸ CFU/mL in the fermentation product after 24 hours.

2. The fermented milk rich in Bifidobacterium longum prepared by the present invention is rich in nutrition, good in taste, and the preparation method is simple and easy. The content of active Bifidobacterium longum in the fermented milk is high, and it can effectively regulate the intestinal flora after eating The probiotic functions such as balance and intestinal adjustment have great market application potential and economic value.

3. The combination of the two kinds of bacteria in the fermented milk constructs a fermentation system based on probiotic bifidobacteria in the milk medium, which solves the problem of the difficulty in the growth of such probiotics in milk. The technical development on this basis is expected to Create a new fermented milk system based on probiotics.

Biological material deposit information:

The Leuconostoc mesenteroides BD01710 strain provided by the present invention has been preserved in the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Management Committee (CGMCC) on August 13, 2012, and the preservation address is Beichen West Road, Chaoyang District, Beijing No. 1 Courtyard No. 3, Zip Code: 100101. The preservation number of the strain is: CGMCC No.6432.

Description of drawings

Fig. 1 is a comparison chart of the growth and curdling of milk in which Bifidobacterium longum and BD01710 are simultaneously added and BD01710 is not added in the present invention.

Detailed ways

The present invention will be described in further detail below by way of examples. If not specified, various raw materials used in the present invention can be purchased from the market. Steps not specifically described are conventional preparation methods in the art.

A method for preparing fermented milk rich in Bifidobacterium longum, the preparation method comprising the following steps:

(1) Inoculate the raw milk with Bifidobacterium longum and Leuconostoc mesenteroides BD01710 respectively, stir and mix evenly, and ferment at a temperature of 30-37°C until the titrated acidity is 70- 95°T, get fermented milk;

(2) Cool the fermented milk prepared in step (1) to 16-22° C. by stirring in an ice bath or by turning over plates to obtain cooled fermented milk;

(3) Filling the cooled fermented milk into a packaging container, then transferring it to an environment with a temperature of 2-10° C. for refrigeration and then ripening for 12-24 hours to obtain fermented milk rich in Bifidobacterium longum.

The Leuconostoc mesenteroides described in step (1) is Leuconostoc mesenteroides BD01710 (Leuconostoc mesenteroides) BD01710 (Leuconostoc mesenteroides) BD01710 of the preservation number CGMCC NO. Can refer to the Chinese patent whose notification number is CN 103013891B), hereinafter referred to as BD01710.

The Bifidobacterium longum and Leuconostoc enterococcus BD01710 in the raw materials for preparation can all be freeze-dried concentrated bacterial powder, which does not affect the scope of protection of the present invention.

The invention utilizes the growth-promoting effect of Leuconostoc mesenteroides on probiotics to realize the growth of probiotics in food media, especially the growth and curdling in milk media, through the growth-promoting effect of Leuconostoc mesenteroides BD01710 A probiotic fermented product based on probiotic fermentation and with a high number of viable bacteria is prepared. The used Leuconostoc mesenteroides subsp. mesenteroides belongs to Leuconostoc mesenteroides subsp. mesenteroides which is recognized as safe.

Leuconostoc enterolis BD01710 is one of the safe edible strains listed by the Ministry of Health of my country (Ministry of Health Announcement No. 8 in 2012) and the US Food and Drug Administration (FDA, 1989). The added Leuconostoc enterococci BD01710 can produce dietary fibers such as dextran and fructan (ZL 201310443498.4). These polysaccharides themselves can be used as healthy dietary supplements, and at the same time bring additional texture benefits to the above-mentioned fermented products. Advantage.

Further, in step (1), the inoculum amount of the Bifidobacterium longum is 10 ⁶ -10 ⁷ CFU/mL, and the inoculum amount of Leuconostoc enterolis BD01710 is 10 ⁶ -10 ⁷ CFU/mL; Or the ratio of the inoculum amount of Bifidobacterium longum to Leuconostoc enterococcus is 1:10-10:1.

Further, in step (1), the raw milk is a conventional choice in the field, consisting of 5-10% whole milk powder or skim milk powder, 0-5% sucrose and water supplemented to 100%, after Prepared after dissolving, mixing, homogenizing, sterilizing and cooling, preferably 10% whole milk powder or skim milk powder, 5% sucrose and water to 100%, preheating the mixed raw materials , homogenize at 60-70°C and 18-22MPa, then heat preservation and sterilization at 90-95°C for 5-10 minutes, and then cool to 40-45°C to obtain raw milk; the percentages are mass percentages.

Or the raw milk is prepared by adding 3-8% sucrose and 0-1.5ppm manganese gluconate after standardizing the raw milk, after dissolving, mixing, homogenizing, sterilizing and cooling, preferably 5% sucrose and 1.0ppm manganese gluconate, mix the raw materials evenly, preheat and heat up, homogenize at 60-70°C and 18-22MPa, then heat and sterilize at 90-95°C for 5-10min, then cool to 35-42 °C, ready to use; the stated percentages are mass percentages. Raw milk or use other dairy ingredients suitable for fermented dairy products.

The present invention also provides fermented milk rich in Bifidobacterium longum prepared by the above-mentioned preparation method.

Example 1

Raw material preparation: raw milk, consisting of 10% whole milk powder or skim milk powder, 5% sucrose and water supplemented to 100%, fully dissolved, mixed evenly, homogeneous (60°C, 22MPa), sterilized at 95°C 5min, cooled to 40°C for use.

(1) Inoculate Bifidobacterium longum (10 ⁷ CFU/mL) and Leuconostoc enterococcus BD01710 (10 ⁶ CFU/mL) into raw milk respectively, stir and mix evenly, and ferment statically at a temperature of 30°C until titration The acidity is 70°T to obtain fermented milk;

(2) Cool the fermented milk prepared in step (1) to 16° C. by stirring in an ice bath or by turning over plates to obtain cooled fermented milk;

(3) Fill the cooled fermented milk into a packaging container, then transfer it to an environment with a temperature of 2° C. for refrigeration and then ripen for 12 hours to obtain fermented milk rich in Bifidobacterium longum.

The above-mentioned fermented milk samples were sampled and counted by plate counting method after gradient dilution. Compared with only inoculating Lactobacillus plantarum ST-III under the same conditions, the results are shown in Table 1, the content of active Lactobacillus plantarum in the fermented milk is higher than 5×10 ⁸ CFU/mL.

Table 1: pH value of fermented milk rich in Bifidobacterium longum and number of live bacteria in fermented milk

Example 2

Raw material preparation: Raw milk consists of 10% whole milk powder or skim milk powder, 5% sucrose and water supplemented to 100%, fully dissolved, mixed evenly, homogeneous (70°C, 22MPa), sterilized at 95°C 10min, cooled to 45°C for use.

(1) Inoculate Bifidobacterium longum (10 ⁶ CFU/mL) and Leuconostoc enterococcus BD01710 (10 ⁷ CFU/mL) into raw milk respectively, stir and mix evenly, and ferment statically at 37°C until titration The acidity is 95°T to obtain fermented milk;

(2) cooling the fermented milk prepared in step (1) to 22° C. by stirring in an ice bath or by turning over plates to obtain cooled fermented milk;

(3) Filling the cooled fermented milk into a packaging container, and then transferring it to an environment with a temperature of 10° C. for refrigerating and then ripening for 24 hours to obtain fermented milk rich in Bifidobacterium longum.

The above-mentioned fermented milk samples were sampled and counted by plate counting method after gradient dilution. Compared with the inoculation of Lactobacillus plantarum ST-III under the same conditions, the results are shown in Table 2. It can be seen that the content of active Bifidobacterium longum in the fermented milk is higher than 5×10 ⁸ CFU/mL.

Table 2: pH value of fermented milk rich in Bifidobacterium longum and number of live bacteria in fermented milk

Example 3

Raw material preparation: Raw milk consists of 10% whole milk powder or skim milk powder, 5% sucrose and water supplemented to 100%, fully dissolved, mixed evenly, homogeneous (70°C, 22MPa), sterilized at 95°C 10min, cooled to 45°C for use.

(1) Inoculate Bifidobacterium longum (10 ⁷ CFU/mL) and Leuconostoc enterococcus BD01710 (3×10 ⁶ CFU/mL) into the raw milk respectively, stir and mix evenly, and ferment at a temperature of 37°C When the titrated acidity is 95°T, fermented milk is obtained;

(2) cooling the fermented milk prepared in step (1) to 22° C. by stirring in an ice bath or by turning over plates to obtain cooled fermented milk;

(3) Filling the cooled fermented milk into a packaging container, and then transferring it to an environment with a temperature of 10° C. for refrigerating and then ripening for 24 hours to obtain fermented milk rich in Bifidobacterium longum.

The above-mentioned fermented milk samples were sampled and counted by plate counting method after gradient dilution. Compared with the inoculation of Lactobacillus plantarum ST-III under the same conditions, the results are shown in Table 3. It can be seen that the content of active Bifidobacterium longum in the fermented milk is higher than 5×10 ⁸ CFU/mL.

Table 3: pH value of fermented milk rich in Bifidobacterium longum and number of live bacteria in fermented milk

Example 4

Raw material preparation: After the raw milk is standardized, add 5% sucrose and 1.0ppm manganese gluconate, mix the raw materials evenly, preheat and heat up, homogenize at 60°C and 18MPa, and then keep warm at 90°C Sterilize for 10 minutes, then cool to 42°C.

(1) Inoculate Bifidobacterium longum (10 ⁷ CFU/mL) and Leuconostoc enterococcus BD01710 (10 ⁶ CFU/mL) into raw milk respectively, stir and mix evenly, and ferment statically at a temperature of 30°C until titration The acidity is 70°T to obtain fermented milk;

(2) Cool the fermented milk prepared in step (1) to 16° C. by stirring in an ice bath or by turning over plates to obtain cooled fermented milk;

(3) Filling the cooled fermented milk into a packaging container, and then transferring it to an environment with a temperature of 2-10° C. for refrigerating and post-cooking for 12 hours to obtain fermented milk rich in Bifidobacterium longum.

The above-mentioned fermented milk samples were sampled and counted by plate counting method after gradient dilution. Compared with the inoculation of Lactobacillus plantarum ST-III under the same conditions, the results are shown in Table 4. It can be seen that the content of active Bifidobacterium longum in the fermented milk is higher than 10 ⁹ CFU/mL.

Table 4: pH value of fermented milk rich in Bifidobacterium longum and number of live bacteria in fermented milk

Example 5

Raw material preparation: After the raw milk is standardized, add 5% sucrose and 1.0ppm manganese gluconate, mix the raw materials evenly, preheat and heat up, homogenize at 60°C and 18MPa, and then keep warm at 90°C Sterilize for 10 minutes, then cool to 35°C.

(1) Inoculate Bifidobacterium longum (10 ⁶ CFU/mL) and Leuconostoc enterococcus BD01710 (10 ⁷ CFU/mL) into raw milk respectively, stir and mix evenly, and ferment statically at 37°C until titration The acidity is 95°T to obtain fermented milk;

(2) cooling the fermented milk prepared in step (1) to 22° C. by stirring in an ice bath or by turning over plates to obtain cooled fermented milk;

(3) Filling the cooled fermented milk into a packaging container, and then transferring it to an environment with a temperature of 10° C. for refrigerating and then ripening for 24 hours to obtain fermented milk rich in Bifidobacterium longum.

The above-mentioned fermented milk samples were sampled and counted by plate counting method after gradient dilution. Compared with the inoculation of Lactobacillus plantarum ST-III under the same conditions, the results are shown in Table 5. It can be seen that the content of active Bifidobacterium longum in the fermented milk is higher than 10 ⁹ CFU/mL.

Table 5: pH value of fermented milk rich in Bifidobacterium longum and number of live bacteria in fermented milk

Example 6

Raw material preparation: After the raw milk is standardized, add 5% sucrose and 1.0ppm manganese gluconate, mix the raw materials evenly, preheat and heat up, homogenize at 60°C and 18MPa, and then keep warm at 90°C Sterilize for 10 minutes, then cool to 40°C.

(1) Inoculate Bifidobacterium longum (10 ⁷ CFU/mL) and Leuconostoc enterococcus BD01710 (3×10 ⁶ CFU/mL) into the raw milk respectively, stir and mix evenly, and ferment at a temperature of 37°C When the titrated acidity is 95°T, fermented milk is obtained;

(2) cooling the fermented milk prepared in step (1) to 22° C. by stirring in an ice bath or by turning over plates to obtain cooled fermented milk;

(3) Filling the cooled fermented milk into a packaging container, and then transferring it to an environment with a temperature of 10° C. for refrigerating and then ripening for 24 hours to obtain fermented milk rich in Bifidobacterium longum.

The above-mentioned fermented milk samples were sampled and counted by plate counting method after gradient dilution. Compared with the inoculation of Lactobacillus plantarum ST-III under the same conditions, the results are shown in Table 6. It can be seen that the content of active Bifidobacterium longum in the fermented milk is higher than 10 ⁹ CFU/mL.

Table 6: pH value of fermented milk rich in Bifidobacterium longum and number of live bacteria in fermented milk

As shown in Figure 1, for embodiment 1, the fermented milk of Bifidobacterium longum BL bacteria and Leuconostoc enterica BD01710 and the fermented milk not inoculated with Leuconostoc enterica BD01710 were inoculated at the same time, in the fermented milk of embodiment 1 The strain amount of Bifidobacterium longum was significantly increased, and the curdling effect was obvious. The growth of Bifidobacterium longum BL and milk curdling can be realized in the milk medium of the present invention, and after 24 hours of fermentation, the number of viable bifidobacteria can reach 5×10 ⁸ CFU/mL.

It should be understood that after reading the above content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims (8)

1. a kind of preparation method of the fermented milk that is rich in Bifidobacterium longum, is characterized in that, its preparation method comprises the following steps: (1) Inoculate the raw milk with Bifidobacterium longum and Leuconostoc mesenteroides BD01710 respectively, stir and mix evenly, and ferment at a temperature of 30-37°C until the titrated acidity is 70- 95°T, get fermented milk; (2) cooling the fermented milk prepared in step (1) to 16-22° C. to obtain cooled fermented milk; (3) Filling the cooled fermented milk into a packaging container, then transferring it to an environment with a temperature of 2-10° C. for refrigeration and then ripening for 12-24 hours to obtain fermented milk rich in Bifidobacterium longum. 2. the preparation method that is rich in the fermented milk of Bifidobacterium longum as claimed in claim 1, is characterized in that, the Leuconostoc enteritidis described in step (1) is the Leuconostoc enteritidis of preservation number CGMCC NO.6432 Leuconostoc mesenteroides BD01710. 3. The method for preparing fermented milk rich in Bifidobacterium longum as claimed in claim 1, characterized in that, in step (1), the inoculation amount of said Bifidobacterium longum is 10 ⁶ -10 ⁷ CFU/ mL, the inoculum amount of Leuconostoc enterococcus BD01710 is 10 ⁶ -10 ⁷ CFU/mL. 4. the preparation method that is rich in the fermented milk of Bifidobacterium longum as claimed in claim 1, is characterized in that, in step (1), the ratio of the inoculum amount of described Bifidobacterium longum and Leuconostoc enteroxicans 1:10-10:1. 5. the preparation method of the fermented milk that is rich in Bifidobacterium longum as claimed in claim 1, is characterized in that, in step (2), the mode of described cooling is the mode of stirring after the ice bath or turning over the plate sheet The way. 6. the preparation method of the fermented milk that is rich in Bifidobacterium longum as claimed in claim 1 is characterized in that, in step (1), described raw milk is whole milk powder or skim milk by 5-10% Powder, 0-5% sucrose and water supplemented to 100% are prepared after dissolving, mixing, homogenizing, sterilizing and cooling; the stated percentages are mass percentages. 7. the preparation method of the fermented milk that is rich in Bifidobacterium longum as claimed in claim 1 is characterized in that, in step (1), described raw milk is after former milk is standardized, adds 3-8% sucrose and 0-1.5ppm manganese gluconate is prepared after dissolving, mixing, homogenizing, sterilizing and cooling; the stated percentages are mass percentages. 8. A fermented milk rich in Bifidobacterium longum prepared by the preparation method according to any one of claims 1-7.