CN116790450A - Fermenting bacteria agent for improving viable bacteria amount of probiotics, N-15 fermented milk and preparation method - Google Patents
Fermenting bacteria agent for improving viable bacteria amount of probiotics, N-15 fermented milk and preparation method Download PDFInfo
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Abstract
The invention discloses a fermentation inoculant for improving the viable count of probiotics, N-15 fermented milk and a preparation method thereof, and belongs to the field of dairy products. Raw milk, sweetener, resistant dextrin and casein are uniformly mixed, heated, lactase enzyme is added for enzymolysis, and after homogenization, sterilization and cooling are sequentially carried out, lactobacillus casei N1115, streptococcus thermophilus, bifidobacterium animalis subspecies i797, lactococcus lactis JCCC 0036 and lactococcus cremoris JCCC 0035 are inoculated for fermentation, and the N-15 fermented milk is obtained after demulsification. The fermentation inoculant for improving the viable count of the probiotics is suitable for preparing N-15 fermented milk, and the prepared N-15 fermented milk has the effect of relaxing bowel, can keep the viable count of the bifidobacteria in a storage period, and has good taste and flavor.
Description
Technical Field
The invention belongs to the technical field of dairy products, and relates to a fermentation inoculant, in particular to a fermentation inoculant for improving the viable bacteria content of probiotics, N-15 fermented milk and a preparation method.
Background
The most commonly used strains in the fermentation process of yoghurt are streptococcus thermophilus and lactobacillus bulgaricus, and the good fermentation performance and the flavor become the preferred combination of yoghurt starter. However, with the intensive research of intestinal flora and probiotics, ordinary yogurt has failed to meet the health needs of consumers, and therefore, more and more probiotics are added to fermented milk.
Lactobacillus casei N1115 is a probiotic with good probiotic characteristics developed by Junlebao dairy groups, and has multiple functions of regulating metabolism, improving immunity, regulating intestinal tract and the like besides good safety. However, the strain has a strong post-acid, which is a major limitation in the use of fermented milk.
Bifidobacteria are important intestinal beneficial microorganisms, and as physiological beneficial bacteria, the bifidobacteria have various important physiological functions such as biological barrier, nutrition, anti-tumor effect, immunity enhancing effect, gastrointestinal tract function improvement, aging resistance and the like for human health.
However, most of the health effects of the addition of probiotics to human bodies in fermented milk cannot produce a noticeable sense of body in a short time. However, for stable yogurt systems, the addition of probiotics often brings about many other immediate problems, such as acid enhancement after fermentation of the milk, yogurt texture changes, bad aroma and flavor, etc. Meanwhile, the probiotics must exist in a large amount in the quality keeping period, can continue to survive and have activity to exert efficacy, and the activity of the bifidobacteria is fast to decline in the process of yoghurt storage and transportation, so that the physiological function of the probiotics cannot be well exerted.
Disclosure of Invention
The invention aims to provide a fermentation inoculant for increasing the viable bacteria content of probiotics so as to solve the problem of bifidobacterium inactivation in the storage and transportation processes of fermented milk;
the invention further aims at providing the N-15 fermented milk prepared by the fermentation inoculant and the preparation method thereof, and the post-acidification and taste of the lactobacillus casei N1115 fermented milk are optimized while the live bacterial amount of probiotics is kept and the bowel relaxing function is achieved.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
the fermenting microbial agent for improving the living microbial quantity of the probiotics is prepared from the following raw materials in parts by weight: the viable bacteria amount is 2-3×10 11 CFU/g lactobacillus casei N1115 0.001-0.1 parts, viable bacteria amount of 1-2×10 9 CFU/g Streptococcus thermophilus0.001-0.1 part, and viable bacteria amount is 1-2×10 10 CFU/g of Bifidobacterium animalis subspecies lactis i 797.001-0.1 part, and viable count of 1-2×10 9 CFU/g lactococcus lactis JCCC 0036 0.001-0.1 parts and viable count of 1-2×10 9 CFU/g lactococcus cremoris JCCC 0035.001-0.1 parts.
The invention also provides N-15 fermented milk prepared from the fermentation microbial inoculum for improving the live microbial biomass of probiotics, and raw materials for preparing active ingredients of the N-15 fermented milk comprise the following components in parts by weight: 900-950 parts of raw cow milk, 50-90 parts of sweetener, 0-50 parts of resistant dextrin, 0-20 parts of casein, 0.1-10 parts of lactase and the fermentation inoculant for improving the live bacteria amount of probiotics.
As one limitation, the sweetener includes at least one of xylitol, erythritol, mogroside, and stevioside.
The invention also provides a preparation method of the N-15 fermented milk, which comprises the steps of uniformly mixing raw cow milk, sweetener, resistant dextrin and casein, heating, adding lactase for enzymolysis, sequentially homogenizing, sterilizing, cooling, inoculating the fermentation inoculant for improving the viable bacteria amount of the probiotics for fermentation, and demulsifying to obtain the N-15 fermented milk.
As a limitation, the heating is at a temperature of 40-45 ℃.
As another limitation, the enzymolysis time is 20-70min.
As a third limitation, the homogenization is carried out at a primary pressure of 15-20 MPa, a secondary pressure of 3-7 MPa and a temperature of 50-69 ℃.
As a fourth limitation, the sterilization is performed at a temperature of 90-98 ℃ for 300-350s;
the temperature after cooling is 36-40 ℃.
As a fifth limitation, the fermentation is carried out at 36-40 ℃ for 5-10 hours, and the acidity after the fermentation is finished is 70-78 ℃;
the demulsification is carried out by stirring for 5-20min at a rotating speed of 1000-3000 r/min.
As a sixth limitation, the raw milk is pretreated by filtering, purifying milk and sterilizing at 83-137 ℃ for 15-30 s.
In the invention, the following components are added:
lactobacillus casei N1115 is separated from a traditional fermented dairy product of Mongolian nationality of an inner Mongolian autonomous region, and the strain is preserved in the China general microbiological culture Collection center (preservation unit address: north Chen West Lu No.1, no. 3 of the Korean region of Beijing city) on 3 months 17 days of 2011, wherein the preservation number is CGMCC No.4691 and the Latin name is Lactobacillus casei;
the bifidobacterium animalis subspecies i797 is preserved in China general microbiological culture Collection center (preservation unit address: north Chen Xili No.1, 3 of the North Chen West Lu of the Beijing area) by 8 months 20 of 2019, and the preservation number is CGMCC No.18403, and the Latin name is Bifidobacterium animalis subsp.
Lactococcus lactis JMCC0036 is selected from traditional dairy products in the south of Gannan, and the strain is preserved in China general microbiological culture Collection center (preservation unit address: north Chen West Lu No.1, hospita No. 3 in the Korean area of Beijing) in 2022, 1 month and 10 days, with a preservation number of CGMCC No.24285 and Latin name of Lactococcus lactis;
lactococcus cremoris JMCC0035 is separated from traditional dairy products of the southern China, and the strain of the strain is preserved in China general microbiological culture Collection center (preservation Unit address: north Chen West Lu No.1, no. 3 in the Korean area of Beijing) for 2022, 1 month and 10 days, wherein the preservation number is CGMCC No.24284, and the Latin name is Lactococcus cremoris;
streptococcus thermophilus can be any living bacteria with the quantity of 1-2×10 9 Commercial products of CFU/g.
By adopting the technical scheme, compared with the prior art, the invention has the following technical progress:
(1) according to the fermentation inoculant for improving the viable count of the probiotics, different strains are selected for compounding, and the combination is optimized, so that the viable count of the bifidobacteria is kept through the competition antagonism relationship among the strains, the acid problem after lactobacillus casei N1115 ferments milk is reduced, and the flavor of the fermented milk is optimized;
(2) the N-15 fermented milk provided by the invention has the function of relaxing bowel, can bring obvious smooth body feeling of the intestinal tract within 3 days, and simultaneously keeps good taste and flavor;
(3) according to the preparation method of the N-15 fermented milk, provided by the invention, through the pre-lactic acid enzyme enzymolysis process and proper starter strain selection, the fermented milk has good taste, flavor and texture, meanwhile, the high survival rate of bifidobacteria can be maintained within the shelf life, and the preparation process is simple and suitable for industrial production.
The fermentation inoculant for improving the viable count of the probiotics is suitable for preparing N-15 fermented milk, and the prepared N-15 fermented milk has the effect of relaxing bowel, can keep the viable count of the bifidobacteria in a storage period, and has good taste and flavor.
Detailed Description
The invention is further illustrated by the following examples. It should be understood that the described embodiments are only for explaining the present invention and do not limit the present invention.
Several fermented milks were prepared and their relevant properties were verified in this example, comprising the following steps in sequence:
sample 1:
s1, after filtering, purifying milk and sterilizing for 15 seconds at 83 ℃, adding 600kg of raw milk into a blending tank, adding 50kg of xylitol, 50kg of resistant dextrin and 20kg of casein, starting stirring for 20 minutes, adding 300kg of raw milk, uniformly stirring, heating to 45 ℃ in a plate-changing cycle, adding 0.1kg of lactase, standing at 45 ℃, and fully carrying out enzymolysis for 70 minutes;
s2, homogenizing under the conditions that the primary pressure is 17.5MPa, the secondary pressure is 5MPa and the temperature is 65 ℃, sterilizing for 300 seconds at 93 ℃, cooling to 38 ℃, sterilizing a fermentation tank mouth and a strain bag by using 75% alcohol, adding a fermentation fungicide alpha 1, stirring at a low speed for 30 minutes, closing stirring, preserving heat and fermenting at 37 ℃, keeping the temperature and fermenting in the middle, keeping the stirring for every 3 hours, recording the fermentation temperature, and after the fermentation is finished, stirring for 5 minutes at the acidity of the fermented milk of 71 ℃ and 1200r/min, and demulsifying to obtain N-15 fermented milk beta 1;
wherein the fermentation bacteria agent alpha 1 is prepared from 100g of viable bacteria with the quantity of 2.5X10 11 CFU/g Lactobacillus casei N1115, 10g viable count of 2×10 9 CFU/g commercial strain Streptococcus thermophilus 1, 10g viable count 2X 10 10 CFU/g bifidobacterium animalis subspecies lactis i797, 10g viable count of 2X 10 9 CFU/g lactococcus lactis JCCC 0036 and 10g viable count of 2×10 9 CFU/g of lactococcus cremoris JCCC 0035.
Samples 2-6:
the amounts of other raw materials and the process parameters of the samples 2-6 are the same as those of the steps S1-S2 in the sample 1, and the difference is only that the fermenting bacteria agents are different, wherein the adding amounts of the fermenting agent strains in the samples 2-6 are as follows: the viable bacteria amount is 2.5X10 11 CFU/g Lactobacillus casei N1115 100g, other strains 10g (wherein the viable count of bifidobacterium animalis subspecies i797 and commercial strains bifidobacterium animalis is 2X 10) 10 CFU/g, the viable count of other strains is 2×10 9 CFU/g), only with the difference that the starter strains are of different species, the specific strain species selection is detailed in table 1:
TABLE 1 species of fermenting bacteria strains in samples 2-6
10 persons of expert sensory evaluation personnel are summoned, and comprehensive evaluation is carried out on the taste favorites of the samples 1-6, and the sensory evaluation method comprises the following steps: the overall taste preference was evaluated on a 9-point intensity scale (9→1 for like→dislike, < 5 for dislike) and the other organoleptic properties on a 10-point intensity scale (10→0 for extremely strong→imperceptible), by which the above fermented milk samples were evaluated for overall preference, aroma, sweetness, sourness, viscosity, and fineness, and the results are shown in table 2:
table 2 sensory evaluation results
Note that: in the table, the same letters represent no significant differences and different letters represent significant differences
As can be seen from table 2, the comparison of sample 1 and sample 3 only differed in that the streptococcus thermophilus strains were different, but the flavor (mainly referring to milk aroma and fermented aroma) and texture (mainly referring to viscosity, powder feel, smoothness, astringency and particulate feel) scores were not significantly affected, indicating that the effect of different streptococcus thermophilus on the fermented milk flavor may not be obvious, and the flavor and texture were mainly provided by virtue of the fermentation process of other strains;
comparing sample 1 with sample 2, the milk fragrance, powder feel and particle feel are all significantly different, which indicates that the commercial strain bifidobacterium animalis and lactococcus lactis are compounded with subspecies lactis, so that the flavor and the texture of the fermented milk are poor;
meanwhile, compared with the sample 2 and the sample 3, the sample 1 has a significant difference in overall preference of the fermented milk, which indicates that the N-15 fermented milk is more popular;
comparing sample 1 and sample 4, the milk aroma and fermentation aroma of sample 4 are significantly affected because of the absence of added lactococcus lactis subspecies lactate and subspecies milk fat;
sample 1 did not have significant differences in scores in terms of overall fermented milk preference, flavor, and texture compared to samples 5 and 6;
in conclusion, the combination of different strains can cause the incompatibility of milk fragrance and fermentation fragrance, so that the overall preference score is affected, and the corresponding effect can be achieved by not simply combining the strains with the same species.
Samples 1-6 were stored at 15 ℃ for 21 days, and the pH, acidity and viable bifidobacterium count of the fermented milk at days 0, 7, 14 and 21 were recorded, respectively, (wherein the acidity measurement is as described in national standard GB 5413.34-2010), and the results are shown in table 3:
TABLE 3 acidity, pH and bifidobacterium viable count test results
As can be seen from Table 3, after 21 days of storage at 15 ℃, the number of viable bifidobacteria in samples 1-4 could meet the national standard requirement of 1.00E+06, while the number of viable bifidobacteria in samples 5 and 6 could not meet the standard requirement, wherein the number of viable bifidobacteria in sample 1 remained the best, probably because lactococcus lactis JCCC 0036 and lactococcus cremoris JCCC 0035 could continue to breathe at 15 ℃, thereby consuming oxygen, and the lower oxygen concentration could maintain the viable bifidobacteria;
at the same time, sample 1 showed better pH and acidity measured during shelf life, indicating better acid production thereafter, probably because lactose in raw milk was enzymatically degraded to glucose and galactose during the preparation process, which can be more rapidly utilized by Streptococcus thermophilus, lactococcus lactis JCCC 0036, lactococcus cremoris JCCC 0035 and bifidobacterium animalis subspecies i797, which strains had much weaker acid production capacity than Lactobacillus casei N1115, and therefore this competition reduced the post-acid production of the fermented milk product.
300 persons were enrolled, of which ages 25-45 years (45% 25-35 years old and 55% 36-45 years old) and 20% male and 80% female were equally divided into A, B, C groups of 100 persons each, wherein consumers with gastrointestinal problems (such as dyspepsia, gastrointestinal motility insufficiency, gastrointestinal sensitivity diarrhea, constipation, etc.) were group a 72, group B62 and group C71 respectively, and fermented milks of sample 1, sample 5 and sample 6 with better taste acceptability were drunk at a frequency of 1 bottle/day, and market research was conducted on them, and the results are shown in table 4, table 5:
table 4 results of efficacy perception by all consumers for samples 1, 5 and 6
Note that: the numbers in the table represent percentages and the letters represent that at 90% confidence there is a significant difference between group A and group B/C
Table 5 results of efficacy perception for consumers with gastrointestinal problems for sample 1, sample 5 and sample 6
Note that: the numbers in the table represent percentages and the letters represent that at 90% confidence there is a significant difference between group A and group B/C
As can be seen from table 4, there are differences in the beneficial effects of probiotics between species, different strains, and different combinations, and on the third day of drinking, the sample 1 (group a) has a significant difference in the body feel of the parts such as "light body", "pleasant to the mind", "aiding digestion of the intestines and stomach", and "bad breath" with respect to the sample 6 (group C); significant differences in "pleasurable" body feel have occurred relative to sample 5 (group B);
on day 7 of drinking, sample 1 showed significant differences in the various evaluation indexes of "body is light", "intestinal immunity is increased", "mood is pleasant", "vitality is brought to the intestinal tract", "gastrointestinal digestion is assisted", "oral malodor is reduced" relative to sample 6;
as can be seen from table 5, for the consumer with intestinal problems, on day 3, the significant difference of "reduced number of gastrointestinal discomfort" had occurred for sample 1 relative to sample 6.
The steps for preparing N-15 fermented milk β2- β6 in this example are substantially the same as steps S1-S2 for preparing N-15 fermented milk β1 in example 1, except that the amounts of raw materials and the process parameters are different, and the specific process parameters are shown in Table 6:
TABLE 6 list of raw materials and process parameters
The other portions of examples 2 to 6 were the same as in example 1 for preparation of sample 1, and N-15 fermented milk was prepared as β2 to β6.
Claims (10)
1. The fermenting bacteria agent for improving the living bacteria amount of the probiotics is characterized by comprising the following raw materials in parts by weight: the viable bacteria amount is 2-3×10 11 CFU/g lactobacillus casei N1115 0.001-0.1 parts, viable bacteria amount of 1-2×10 9 CFU/g streptococcus thermophilus 0.001-0.1 parts, and viable bacteria amount of 1-2×10 10 CFU/g of Bifidobacterium animalis subspecies lactis i 797.001-0.1 part, and viable count of 1-2×10 9 CFU/g lactococcus lactis JCCC 0036 0.001-0.1 parts and viable count of 1-2×10 9 CFU/g lactococcus cremoris JCCC 0035.001-0.1 parts.
2. An N-15 fermented milk prepared from the fermenting agent for increasing the viable bacteria amount of probiotics according to claim 1, characterized in that raw materials for preparing active ingredients of the N-15 fermented milk comprise, in parts by weight: 900-950 parts of raw cow milk, 50-90 parts of sweetener, 0-50 parts of resistant dextrin, 0-20 parts of casein, 0.1-10 parts of lactase and the fermentation inoculant for improving the live bacteria amount of probiotics.
3. The N-15 fermented milk of claim 2, wherein the sweetener comprises at least one of xylitol, erythritol, mogroside, and stevioside.
4. The method for preparing the N-15 fermented milk according to claim 2 or 3, wherein raw milk, a sweetener, resistant dextrin and casein are uniformly mixed, heated, lactase is added for enzymolysis, homogenization, sterilization and cooling are sequentially carried out, the fermentation inoculant for improving the viable bacteria amount of the probiotics is inoculated for fermentation, and the N-15 fermented milk is obtained after demulsification.
5. The method for producing an N-15 fermented milk according to claim 4, wherein the heating is performed at a temperature of 40 to 45 ℃.
6. The method for producing an N-15 fermented milk according to claim 4, wherein the enzymolysis is performed for 20 to 70 minutes.
7. The method for producing an N-15 fermented milk according to claim 4, wherein the primary pressure is 15 to 20 MPa, the secondary pressure is 3 to 7 MPa, and the temperature is 50 to 69 ℃.
8. The method for producing an N-15 fermented milk according to claim 4, wherein the sterilization is performed at a temperature of 90 to 98℃for 300 to 350 seconds;
the temperature after cooling is 36-40 ℃.
9. The method for producing an N-15 fermented milk according to claim 4, wherein the fermentation is carried out at 36 to 40℃for 5 to 10 hours and the acidity after completion of the fermentation is 70 to 78℃T;
the demulsification is carried out by stirring for 5-20min at a rotating speed of 1000-3000 r/min.
10. The method for producing an N-15 fermented milk according to any one of claims 4 to 9, wherein the raw milk is pretreated by filtration, purification and sterilization at 83 to 137 ℃ for 15 to 30 seconds.
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