CN114891703B - Compound leaven with weak post-acidification function, application thereof and preparation method of yogurt - Google Patents
Compound leaven with weak post-acidification function, application thereof and preparation method of yogurt Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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.
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- 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
- A23C9/1236—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using Leuconostoc, Pediococcus or Streptococcus sp. other than Streptococcus Thermophilus; Artificial sour buttermilk in general
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- 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
- A23V2400/137—Delbrueckii
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- 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
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/249—Thermophilus
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- C12R2001/00—Microorganisms ; Processes using microorganisms
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- C12R2001/225—Lactobacillus
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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Abstract
The invention relates to the technical field of microbial fermentation, in particular to a compound leavening agent with weak post-acidification effect, application thereof and a preparation method of yogurt. The composite leaven comprises Streptococcus thermophilus CICC 6038 and Lactobacillus delbrueckii subspecies Bulgaricus CICC 6047, and the ratio of the number of viable bacteria of the Streptococcus thermophilus CICC 6038 to the number of viable bacteria of the Lactobacillus delbrueckii subspecies Bulgaricus CICC 6047 is 10-50: 1, and preferably 19:1. the yogurt prepared by the compound starter has unique fresh and sweet flavor under the condition of no sugar or other sweeteners, is sticky, soft and fine in texture, smooth in mouthfeel, proper in sour taste and strong in milk fragrance. The method has the characteristics of high fermentation speed, high viscosity, typical aroma and weak post-acidification, can be applied to dairy industry to control post-acidification, improve product quality, prolong shelf life and improve the taste of sugar-free or sugar-reduced yogurt.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a composite leavening agent with weak post-acidification effect, application thereof and a preparation method of yogurt.
Background
The yogurt is prepared from raw milk and/or reconstituted milk by sterilizing, inoculating Streptococcus thermophilusStreptococcus thermophilus) And Lactobacillus delbrueckii subspecies bulgaricus (B.delbrueckii)Lactobacillus delbrueckii subsp. bulgaricus) Fermenting to obtain the final product. During the fermentation process, the strains endow the dairy products with unique sensory characteristics and nutritional attributes through complex symbiotic interaction. The starter is a key factor directly determining the final flavor and quality of the fermented dairy product, and the fermentation characteristics depend on the selection of strains and the combination effect. In the process of low-temperature storage and cold-chain transportation of the yogurt, the metabolic activity of the residual starter strains can continuously and slowly produce acid, so that the product defects of over-high acidity, whey precipitation, reduced sensory quality, reduced quantity of live bacteria, pollution of putrefying bacteria and the like can be caused. To maintain shelf-life quality stability and slow down the post-acidification process, manufacturers can use ultra-high pressure (HHP), pulsed electric field, CO 2 The key enzyme activity of the strain is inactivated by means of treatment, post heat treatment and the like, and exogenous substances such as bacteriocin, chitosan, pectin zymolyte, glutamine transaminase and the like can be added for intervention, so that the measures can increase the commercial cost, influence the rheological property and the sensory property of the yogurt product, or reduce the nutritional value of the yogurt product. Screening and developing wild strains of the leavening agent with excellent fermentation performance and weak post-acidification potential is an important strategy for controlling post-acidification from the source and being economical and safe, and simultaneously, the amount of sugar added for balancing sour taste can be reduced.
After the fermentation was completed, the pH dropped by 0.3 unit during the period of refrigeration at 4 ℃ for 21d, and it was considered to be weak post-acid. Although a single strain can have the post-acid inhibition effect, the single strain is difficult to have the comprehensive characteristics of quick acid production, high viscosity production, good flavor production and the like, and does not meet the definition of the yogurt in the domestic and foreign standards.
Therefore, the development of the excellent compound leaven which can solve the common problem in the post-acidification industry and meet the requirements of clean labels and sugar consumption reduction trend has wide application and popularization values.
Disclosure of Invention
In order to solve the technical problems, the invention provides a compound leavening agent with weak post-acidification effect, application thereof and a preparation method of yogurt.
The invention provides a composite leaven with weak post-acidification effect, which comprises 10-50% of viable bacteria number ratio of streptococcus thermophilus CICC 6038 to lactobacillus delbrueckii subspecies bulgaricus CICC 6047 to streptococcus thermophilus CICC 6038 to lactobacillus delbrueckii subspecies bulgaricus CICC 6047: 1, and preferably 19:1.
the invention provides application of the compound starter in preparing yogurt, and the yogurt comprises weak post-acidified whole-fat yogurt and weak post-acidified skim yogurt, and the yogurt is of a set type or a stirring type.
Optionally, the yogurt is yogurt without sucrose.
Optionally, the yogurt is yogurt without sweeteners and/or additives.
The invention provides a preparation method of weak post-acidified yogurt, which at least comprises the following steps:
s1, activated streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies bulgaricus CICC 6047;
s2, sterilizing the raw materials for preparing the yogurt;
s3, enabling streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies Bulgaria CICC 6047 to be mixed according to the ratio of viable bacteria number of 10-50: 1. preferably, 19:1 is inoculated on the raw material;
and S4, fermenting after inoculation to obtain the yogurt product.
Optionally, in S1, the activating comprises:
s11, respectively inoculating frozen streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies bulgaricus CICC 6047 preservation bacteria liquid into a milk test tube, and performing standing culture on the streptococcus thermophilus CICC 6038 at 43 ℃ to obtain a first generation seed liquid and performing anaerobic standing culture on the lactobacillus delbrueckii subspecies bulgaricus CICC 6047 at 43 ℃ to obtain a first generation seed liquid; the culture time is 14-16 hours;
s12, after the culture is finished, respectively transferring streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies Bulgaria CICC 6047 into a cow milk blue-covered bottle, performing static culture on the first generation seed liquid of the streptococcus thermophilus CICC 6038 at 43 ℃ to obtain a second generation seed liquid, and performing anaerobic static culture on the first generation seed liquid of the lactobacillus delbrueckii subspecies Bulgaria CICC 6047 at 43 ℃ to obtain a second generation seed liquid; culturing for 7-9 hours to obtain the second-generation seed liquid, and storing at the temperature of 2-8 ℃.
Optionally, in S2, the sterilizing comprises: taking whole milk or skimmed milk as a raw material, heating the raw material to 58-62 ℃, and homogenizing under the condition of 15-25 MPa; sterilizing for 8-12 minutes at 94-96 ℃.
Optionally, in S3, the inoculation amount of Streptococcus thermophilus CICC 6038 is 5 × 10 6 cfu/mL ~ 5×10 7 The inoculation amount of cfu/mL and Lactobacillus delbrueckii subspecies bulgaricus CICC 6047 is 1 multiplied by 10 6 cfu/mL ~ 5×10 6 cfu/mL。
Optionally, in S3, the fermentation time is 4-7 hours, and the fermentation temperature is 42-44 ℃.
Optionally, in S4, when the pH of the yogurt obtained after fermentation is reduced to 4.6, the fermentation is terminated, and the yogurt is stored at a temperature of 2 to 8 ℃.
Compared with the prior art, the technical scheme provided by the embodiment of the invention has the following advantages:
the yogurt prepared by the composite starter has weak post-acidification effect, the pH value is reduced to be less than 0.3 during the cold storage at 4 ℃ for 50 days, and the pH value of the yogurt is maintained at 4.2-4.4. The yogurt prepared by the composite leavening agent has unique fresh and sweet flavor under the condition of no sugar or other sweeteners, so the yogurt is particularly suitable for preparing the sugar-free yogurt. The yogurt prepared by the compound starter has sticky, soft and fine texture, smooth mouthfeel, proper sour taste and strong milk flavor. Is particularly suitable for preparing yogurt with simplified formula.
The composite leavening agent also has the technical advantage of high fermentation speed. Compared with yogurt prepared from commercial leaven, the yogurt has good organoleptic and texture characteristics.
The composite leaven has symbiotic effect among strains, and the two bacteria compounded according to a specific proportion have the characteristics of high fermentation speed, high viscosity, typical fragrance and weak post-acidification during fermentation, so that the composite leaven can be applied to the dairy industry to control post-acidification, improve the product quality, prolong the shelf life and improve the taste of sugar-free or sugar-reduced yogurt.
Drawings
FIG. 1 is a scanning electron microscope morphology of Streptococcus thermophilus CICC 6038;
FIG. 2 is a scanning electron microscope morphology of Lactobacillus delbrueckii subspecies Bulgaria CICC 6047;
FIGS. 3 to 6 are graphs showing the changes in the micro-rheological properties of the fermentation process in example 7;
FIG. 7 is a radar chart of the response of the electronic nose sensor to the sample in example 8;
fig. 8 is a graph of PCA results of e-nose analysis of flavor profiles in example 8.
Detailed Description
In order that the above objects, features and advantages of the present invention may be more clearly understood, a solution of the present invention will be further described below. It should be noted that the embodiments of the present invention and features of the embodiments may be combined with each other without conflict.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the invention may be practiced otherwise than as described herein; it is to be understood that the embodiments described in this specification are only some embodiments of the invention, and not all embodiments.
The embodiment of the invention provides a composite leavening agent with weak post-acidification effect, which comprises Streptococcus thermophilus CICC 6038 and Lactobacillus delbrueckii subspecies Bulgaricus CICC 6047, wherein the ratio of the number of viable bacteria of the Streptococcus thermophilus CICC 6038 and the Lactobacillus delbrueckii subspecies Bulgaricus CICC 6047 is 10-50: 1, preferably 10-20: 1 when the two strains compounded according to the specific proportion are used for preparing the yogurt, the weak post-acidification effect is achieved, the pH value is reduced to be less than 0.3 during the cold storage at 4 ℃ for 50 days, the pH value of the yogurt can be maintained to be 4.2-4.4, if the pH value is too low, the taste is too acid, and if the pH value is too high, the sour taste is insufficient, and the taste of the yogurt is not good. The subsequent weak acidification can control the post-acidification, improve the product quality and prolong the shelf life. In order to further improve the fermentation efficiency of the composite leavening agent and shorten the fermentation time, the ratio of the number of viable bacteria of streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subsp bulgaricus CICC 6047 is 18-20: 1, more preferably 19:1. tests show that the composite leaven in the range can obviously shorten the fermentation time and has the technical advantage of high fermentation speed. Within the optimal proportion range, the prepared yogurt has good taste and texture, the sensory performance is further improved, and the yogurt has unique fresh and sweet flavor and strong milk flavor.
The embodiment of the invention also provides an application of the modified compound starter in preparing yogurt, and the yogurt comprises weak post-acidified full-fat yogurt and weak post-acidified skim yogurt. The compound leaven provided by the embodiment of the invention has good mouthfeel, texture and flavor when being used for preparing full-cream yogurt, and also has good mouthfeel, texture and flavor when being prepared by taking defatted and semi-defatted dairy products as raw materials. Therefore, the method is particularly suitable for preparing the defatted and semi-defatted yogurt.
As an improvement of the technical scheme of the embodiment of the invention, the yogurt obtained by fermenting the compound fermentation agent of the embodiment of the invention has unique sweet flavor, low acidity and very high palatability without adding sucrose, and is particularly suitable for preparing sugar-free yogurt. And other sweeteners are not required to be added.
As an improvement of the technical scheme of the embodiment of the invention, the yogurt obtained by fermenting the compound starter of the embodiment of the invention has good mouthfeel and texture without adding other additives, such as a coagulant, a stabilizer, a thickener, a sweetener and the like, so the yogurt is particularly suitable for the preparation of the yogurt without adding additives, and is particularly suitable for the market demand of the yogurt with simplified formula and cleanness at present.
The embodiment of the invention provides a preparation method of weak post-acidified yogurt, which at least comprises the following steps:
s1, activated streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies bulgaricus CICC 6047;
s2, sterilizing the raw materials for preparing the yogurt;
s3, performing pretreatment on streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subsp bulgaricus CICC 6047 according to the ratio of viable bacteria number of 10-50: 1. preferably, the ratio of 19:1 is inoculated on the raw material;
and S4, fermenting after inoculation to obtain the yogurt product.
As an improvement of the technical solution of the embodiment of the present invention, in S1, the activating includes:
s11, respectively inoculating frozen streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies Bulgaria CICC 6047 preservation bacteria liquid into a milk test tube, performing static culture on the streptococcus thermophilus CICC 6038 at 42-44 ℃ and preferably 43 ℃ to obtain a first-generation seed liquid, and performing anaerobic static culture on the lactobacillus delbrueckii subspecies Bulgaria CICC 6047 at 42-44 ℃ and preferably 43 ℃ to obtain a first-generation seed liquid; the culture time is 14-16 hours;
s12, after the culture is finished, respectively transferring streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies Bulgaria CICC 6047 into a cow milk blue-covered bottle, performing static culture on the first generation seed liquid of the streptococcus thermophilus CICC 6038 at 42-44 ℃ and preferably 43 ℃ to obtain a second generation seed liquid, and performing anaerobic static culture on the first generation seed liquid of the lactobacillus delbrueckii subspecies Bulgaria CICC 6047 at 42-44 ℃ and preferably 43 ℃ to obtain a second generation seed liquid; culturing for 7-9 hours to obtain 2-generation seed liquid, and storing at the temperature of 2-8 ℃.
As an improvement of the technical solution of the present invention, in S2, the sterilization includes: taking whole milk or skimmed milk as a raw material, heating the raw material to 58-62 ℃, preferably 60 ℃, and homogenizing under the condition of 15-25 MPa, preferably 20 MPa; sterilizing at 94-96 deg.C, preferably 95 deg.C for 8-12 min, preferably 10 min.
As an improvement of the technical scheme of the invention, in S3, the inoculation amount of streptococcus thermophilus CICC 6038 is 5 multiplied by 10 6 cfu/mL ~ 5×10 7 The inoculation amount of cfu/mL and Lactobacillus delbrueckii subspecies bulgaricus CICC 6047 is 1 multiplied by 10 6 cfu/mL ~ 5×10 6 cfu/mL。
As an improvement of the technical scheme of the invention, in S3, the fermentation time is 4-7 h, and the fermentation temperature is 42-44 ℃, preferably 43 ℃.
As an improvement of the technical scheme of the invention, in S4, when the pH value of the yogurt obtained after fermentation is reduced to 4.6, the fermentation is terminated as a fermentation end point, and the yogurt is stored at the temperature of 2-8 ℃.
The specific embodiment is as follows:
example 1 identification of CICC 6038
(1) The CICC 6038 is subjected to scanning electron microscope observation (shown in figure 1) and physiological and biochemical determination. CICC 6038 bacterial colony is grey white, the thallus is in an ellipsoid shape, the size of the thallus is 0.5-0.7 mu m multiplied by 0.8-1.4 mu m, and the thallus is arranged singly, in pairs or in a chain shape. Glucose and sucrose are fermented, and maltose and raffinose are not fermented.
(2) A bacteria average nucleotide consistency (ANI) identification and detection method is adopted, and the ANI value is 95-96% and can be used as a threshold value standard defined by prokaryote species. Determination of the CICC 6038 genomic sequence, CICC 6038 andStreptococcus thermophilusNCTC 12958 T the ANI value of (b) was 98.36%. CICC 6038 is identified asStreptococcus thermophilus。
Example 2 identification of CICC 6047
(1) The CICC 6047 is subjected to scanning electron microscope observation (shown in figure 2) and physiological and biochemical determination. The CICC 6047 thalli are rod-shaped, 0.5-0.7 mu m is multiplied by 2.6-4.7 mu m, and are arranged singly or in pairs. Fermenting glucose, lactose and fructose without using sucrose, starch, xylose and arabinose.
(2) A bacteria average nucleotide consistency (ANI) identification and detection method is adopted, and the ANI value is 95-96% and can be used as a threshold value standard defined by prokaryote species. Determination of the sequence of the CICC 6047 genome, CICC 6047 andLactobacillus delbrueckii subsp. bulgaricusATCC 11842 T the ANI value of (A) was 99.28%. CICC 6047 is identified asLactobacillus delbrueckii subsp. bulgaricus。
Example 3 preparation of yogurt Using Complex strains
The preparation method comprises the following steps:
(1) Strain activation: inoculating 200 mu L of the frozen and preserved bacterium liquid into a 5 mL cow milk test tube, performing static culture on streptococcus thermophilus CICC 6038 at 43 ℃, and performing anaerobic static culture on lactobacillus delbrueckii subspecies Bulgaria CICC 6047 at 43 ℃. After culturing for 15 hours, each test tube is respectively transferred to a 100 mL cow milk blue cap bottle, culturing for 8 hours under the same condition to obtain second-generation seed liquid, and storing at low temperature of 4 ℃ for later use.
(2) Homogenizing and sterilizing: taking whole/skimmed milk as a raw material, heating the milk to 60 ℃, and homogenizing under the condition of 20 MPa; sterilizing at 95 deg.C for 10 min.
(3) Inoculating and fermenting: after sterilization, the temperature is reduced to 43 ℃. With a CICC 6038: the viable count of CICC 6047 =19:1 proportion inoculation, the inoculation amount of Streptococcus thermophilus CICC 6038 is 5 multiplied by 10 6 cfu/mL~5×10 7 The inoculation amount of cfu/mL and Lactobacillus delbrueckii subspecies Bulgaria CICC 6047 is 1 multiplied by 10 6 cfu/mL~5×10 6 cfu/mL, the fermentation time is 4-7 hours, the fermentation is stopped when the pH value of the fermented milk is reduced to 4.6, and the fermented milk is refrigerated at 4 ℃.
Example 4 pH Change in set fermented milks during fermentation and storage at Low temperatures
Grouping experiments: grouped as in table 1.
Table 1: experimental groups for pH determination during fermentation and storage
The fermentation method of samples 1 and 2 was the same as in example 3.
The samples 3 and 4 were inoculated with 1X 10 of an imported commercial weak post-acidification starter 7 cfu/mL~5×10 7 cfu/mL, other fermentation conditions were the same as in example 3.
The total amount of inoculation in samples 5 to 13 was the same as in example 3, and the other fermentation conditions were the same as in example 3.
pH during fermentation of fermented milk was continuously monitored using an iCinac fermentation monitor, and pH changes of fermented milk stored at 4 ℃ for 1d, 10 d, 20 d, 30 d, 40 d and 50 d were measured using a pH meter, and the results are shown in Table 2.
Table 2: time to fermentation end and pH change of fermented milk at 4 deg.C
From the viewpoint of weak post-acidification, as shown in Table 2, each of sample 1, sample 2, sample 8 and sample 9 had a weak post-acidification effect, and the pH change of the fermented yogurt was substantially stable up to 50 days under refrigeration. Wherein, the post-acidification degree of the sample 1 and the sample 2 is weaker than that of the sample 3 and the sample 4, and the weak post-acidification effect is better.
The pH of samples 10 and 11 was maintained above 4.4 during refrigeration, although weak post-acidification was evident, the fermentation effect was poor; the post-acidification of samples 12 and 13 was significant during storage, and the resulting yoghurts had a sour mouthfeel and a bad taste. Compared with the prior art, the compound strain has a synergistic effect in the fermentation process, and the post-acidification effect of the CICC 6047 is effectively weakened by the addition of the CICC 6038 during the storage period.
From the perspective of fermentation time, the yogurt of samples 1-4 all reached the fermentation end within 5 hours, wherein the time for sample 1 and sample 2 to reach the fermentation end is faster than the time for sample 3 and sample 4 to reach the fermentation end by about 20 minutes. The curd time for samples 8 and 9 was longer than for samples 1 and 2. In samples 5 to 9, the fermentation time of sample 9 is the longest, and the fermentation time of sample 5 is the shortest. The time for reaching the end point of the CICC 6038 single-strain fermented yogurt (sample 10 and sample 11) is more than 9 hours, the time for reaching the end point of the CICC 6047 single-strain fermented yogurt (sample 12 and sample 13) is more than 5.5 hours, and the fermentation efficiency is lower than that of double-strain composite fermentation.
Example 5 sensory evaluation of set fermented milk
Grouping experiments: the grouping is performed in the same manner as in embodiment 4.
With reference to the industry standard of RHB 104-2020 sensory evaluation rules for fermented milk, the fermented milk was evaluated after 1 day of after-ripening at 4 ℃, and 10 volunteers formed a sensory evaluation panel, and the panel received two 2-hour training sessions, requiring evaluation of the appearance, odor, flavor and texture of the fermented milk. In each training process, four domestic famous fermented milks are respectively provided for sensory evaluation panelists to help the panelists to define and refer to various indexes of the fermented milks. The evaluation criteria are shown in table 3. The scores of 10 volunteers were statistically calculated as average scores, and the results are shown in Table 4.
Table 3: sensory evaluation system evaluation standard of fermented milk
Table 4: fermented milk sensory evaluation result statistical table
Table 4 shows that the flavor of the single-strain CICC 6038 fermented milk was insufficient, and that the tissue state of the single-strain CICC 6047 fermented milk was slightly inferior and a small amount of whey was precipitated. Compared with the prior art, the two strains are mutually influenced through various complex interaction relations during compound fermentation, and the acid production speed is high, the characteristic flavor is rich, the viscosity is good, and the like. In the yogurt prepared by compounding and fermenting the CICC 6038 and the CICC 6047 in different proportions, the ratio of (19): the total score of 1 proportion is the highest, the total score is superior to yogurt in other proportions in color, taste, smell and tissue state, and the organoleptic properties are close to those of the imported commercial leavening agent.
Example 6 texture Properties of set fermented milk
Experimental grouping: grouped according to table 5.
Table 5: texture property experimental grouping of fermented milk
The experimental method comprises the following steps: a TA-XT2 texture tester is adopted to measure texture characteristics, a 0.5-inch cylindrical non-metal probe is arranged on the texture tester, a prepared yogurt sample is placed under the probe, software is started, the probe can move downwards at a certain speed, when the probe is pressed downwards until the gel surface is just broken (formed), the software automatically records the stress condition at the moment, and the peak value is gel strength. Gel break distance is the distance the probe moves down when the gel breaks during the test and can be used to characterize elasticity. The experimental results are as follows: see table 6.
Table 6: results of texture Properties of fermented milk
From the results shown in table 6, it was found that CICC 6038 and CICC 6047 are expressed at 19:1, the obtained full-cream yogurt and the skim yogurt have texture slightly superior to that of a commercial starter.
Example 7 micro-rheological Properties of set-type fermented milk
Experimental grouping: the experimental groups were as in example 6.
The experimental method comprises the following steps: a Rheolaser Master micro-rheometer was used to take 20 mL of the milk sample inoculated with the starter, and the milk sample was poured into a glass tube dedicated to the sterile micro-rheometer, which was quickly transferred to a measuring tank set at 43 ℃. Measurements were taken every 1 min to the end of fermentation. The glass bottle was immediately removed and placed in a refrigerated environment at 4 ℃ for 24 h, and then the sample was returned to room temperature (25 ℃) and placed in the assay chamber for further data collection. The collection and analysis of the raw data is performed in software on the instrument.
Results and analysis: the changes of the flow Factor (FI), the elastic factor (EI), the Macroscopic Viscosity (MVI) and the solid-liquid equilibrium value (SLB) in the fermentation process are shown in FIGS. 3-6.
The FI reaction microscopic particles move fast and slow in the sample, and the larger the value of the FI reaction microscopic particles is, the faster the particles move, the more unstable the structure is, and the lower the viscosity of the sample is. As shown in FIG. 3, in the first 2 to 3 hours, the FI values of all groups of samples fluctuate in a relatively high range, the FI values of the samples 3 and 4 enter an inflection point first to form a gel structure, and the samples enter a stable state in about 3.2 hours. Sample 1 and sample 2 entered the inflection point at around 3.2 hours and entered steady state at around 3.5 hours.
EI is proportional to the modulus of elasticity of the sample, with a larger value indicating a greater elasticity of the sample. As shown in FIG. 4, the EI values of the samples of each group were fluctuated in a relatively low range 3 hours before the start of the fermentation, and the EI values of sample 3 and sample 4 started to rise first near 3 hours, and the EI values of sample 1 and sample 2 started to rise straight around 3.2 hours after the start of the fermentation, reached the maximum value around 4.2 hours, and then slowly declined thereafter.
MVI is directly proportional to the macroscopic viscosity of the sample. As shown in FIG. 5, the MVI values of the samples of each group fluctuate within a relatively low range from the beginning of fermentation to about 3.4 hours, and the MVI values of the samples start to gradually increase after 3.6 hours, the viscosity of the samples gradually increases, and the viscosity of sample 1 is higher than that of sample 3 and the viscosity of sample 4 is higher than that of sample 2 at the later stage of fermentation.
SLB is proportional to the viscoelastic properties of the sample, which is a ratio of the properties of the solid to the properties of the liquid in the sample. When 0 < SLB.ltoreq.0.5, the sample shows elastic/solid properties; a SLB < 1 of 0.5. Ltoreq. Indicates that the sample exhibits viscous/liquid properties. As shown in FIG. 6, the SLB values in the first 3 hours were in an irregular fluctuation state, and then the SLB values in the samples of each group were linearly decreased, at which time the samples reached the gel point and the samples began to change from a liquid property to a partial solid property. The SLB values of the samples of each group reached a steady state 4 hours after the start of fermentation, with the SLB value of sample 1 being greater than that of sample 3 at the steady state, indicating that sample 1 is more biased toward the solid state; sample 2 and sample 4 were close.
Example 8 flavor Profile analysis
Grouping experiments: grouped according to table 7.
Table 7: experimental group of volatile flavor of fermented milk
The experimental method comprises the following steps: the electronic nose is an effective technology for rapid classification and discrimination, and a FOX 4000 electronic nose system is adopted for determination, wherein the electronic nose system is provided with three sensor matrix chambers and 17 metal oxide sensors.
The instrument setting parameters are as follows: (1) signal acquisition: the acquisition time is 120 seconds; the data acquisition period is 1 second; data acquisition delay 1080 seconds; the flow rate is 150 mL/min; (2) sample injection: sampling amount is 5000 muL; the injection speed is 500 muL/s; (3) a heater: incubation period 300 seconds; the incubation temperature is 60 ℃; (4) heating the oscillator: stirring speed 500 rpm; starting stirring for 5 seconds; stopping stirring for 5 seconds; (5) a sample injection needle: the cleaning time is 120 seconds; the injector temperature is 70 ℃; fill speed 500 muL/sec.
Results and analysis:
(1) Analysis of sensor response to sample
The mean of the response extrema of the electronic nose sensor to the 6 groups of samples is shown in table 8.
Table 8: mean value of extreme values of response of electronic in-nose sensor to fermented milk sample
Note: the difference in lowercase letters in the same row indicates that there is a significant difference between samples (P < 0.05)
The response values of the six sensors, namely LY2/G, LY2/AA, LY2/gCTL, T30/1, T70/2 and T40/2, of the sample No. 1 are obviously different from those of the sample No. 3, the sample No. 1 shows higher response values on the three sensors, namely LY2/G, LY2/AA and LY2/gCTL, and the response values of the other 11 sensors are in a steady change trend and have smaller fluctuation and difference.
Sample No. 2 and sample No. 4 showed significant differences in sensors LY2/G, LY2/AA, LY2/gCTL, LY2/gCT, T30/1, P10/2, T70/2, PA/2, P30/1, P40/2, P30/2, T40/2, and sample No. 2 showed higher response values in sensors T30/1, P10/2, T70/2, PA/2, P30/1, P40/2, P30/2, T40/2, wherein the difference in the extreme mean values of the response intensities of the sensors P30/1 was the greatest. In combination with the sensor response characteristics with higher response values in Table 8, it can be inferred that CICC 6038 + CICC 6047 fermented and defatted yogurt is rich in more organic compounds.
(2) Radar chart analysis
Radar mapping analysis is an important multivariate analysis method, which is widely used in e-nose related research and can reveal the overall and individual differences of the sensor array in response intensity to different sample signals. Radar plots of the response of 17 sensors in the FOX 4000 electronic nose system to fermented milk are shown in fig. 7.
From FIG. 7, it can be observed that the 6 samples were divided into 2 groups, i.e., one group of No. 14 and No. 15, and the other group of No. 1-4, indicating that there was a significant difference in flavor profile between the milk before and after fermentation; the overall flavor profiles of fermented whole milk and fermented skim milk are similar, and it is presumed that they have similar flavor fermentation production performance.
(3) Principal component analysis
The Principal Component Analysis (PCA) is to reduce the dimension of the original data into two new index principal components 1 (PC 1) and 2 (PC 2) by an algorithm, which are displayed on the graph as X-axis and Y-axis respectively. Principal Component Analysis (PCA) was performed on a matrix of 17 sensor signal response intensity extrema collected from 6 sets of samples, and fig. 8 is the electronic nose PCA result.
As can be seen in fig. 8, the contribution rates of PC-1 and PC-2 to the overall flavor difference were 99.923% and 0.0649%, respectively, which can represent the original information of the sample. The main component analysis shows that independent areas formed by the fermented milks are not overlapped, which indicates that the fermented milks CICC 6038 + CICC 6047 have unique flavor characteristics.
The above description is merely illustrative of particular embodiments of the invention that enable those skilled in the art to understand or practice the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. The application of the composite starter with weak post-acidification effect in preparing yoghurt is characterized in that the composite starter comprises Streptococcus thermophilus CICC 6038 and Lactobacillus delbrueckii subsp. 1.
2. Use according to claim 1, wherein the yogurt comprises a weak post-acidified full fat yogurt and a weak post-acidified skim yogurt, the yogurt being of the set or stirred type.
3. Use according to claim 2, wherein the yogurt is a yogurt without added sucrose.
4. Use according to claim 2 or 3, wherein the yogurt is yogurt without the addition of additives, said additives comprising sweeteners.
5. A method for preparing weak post-acidified yogurt, characterized by comprising at least the following steps:
s1, activated streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies bulgaricus CICC 6047;
s2, sterilizing the raw materials for preparing the yogurt;
s3, adding Streptococcus thermophilus CICC 6038 and Lactobacillus delbrueckii subsp. Bulgaricus CICC 6047 in a ratio of viable bacteria number of 18-20: 1 is inoculated on the raw material;
and S4, fermenting after inoculation to obtain the yogurt product.
6. The method according to claim 5, wherein in S1, the activating comprises:
s11, respectively inoculating frozen streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies bulgaricus CICC 6047 preservation bacteria liquid into a milk test tube, and performing static culture on the streptococcus thermophilus CICC 6038 at 42-44 ℃ to obtain a first generation seed liquid and performing anaerobic static culture on the lactobacillus delbrueckii subspecies bulgaricus CICC 6047 at 42-44 ℃ to obtain a first generation seed liquid; the culture time is 14-16 hours;
s12, after the culture is finished, respectively transferring streptococcus thermophilus CICC 6038 and lactobacillus delbrueckii subspecies bulgaricus CICC 6047 into a cow milk blue cover bottle, performing static culture on a first generation seed solution of streptococcus thermophilus CICC 6038 at 42-44 ℃ to obtain a second generation seed solution, and performing anaerobic static culture on a first generation seed solution of lactobacillus delbrueckii subspecies bulgaricus CICC 6047 at 42-44 ℃ to obtain a second generation seed solution; culturing for 7-9 hours to obtain the second-generation seed liquid, and storing at the temperature of 2-8 ℃.
7. The method of manufacturing according to claim 5, wherein in S2, the sterilizing includes: taking whole milk or skimmed milk as a raw material, heating the raw material to 58-62 ℃, and homogenizing under the condition of 15-25 MPa; sterilizing for 8-12 minutes at 94-96 ℃.
8. The method according to claim 5, wherein the amount of S3 inoculated with S.thermophilus CICC 6038 is 5X 10 6 cfu/mL ~ 5×10 7 The inoculation amount of cfu/mL and Lactobacillus delbrueckii subspecies bulgaricus CICC 6047 is 1 multiplied by 10 6 cfu/mL ~ 5×10 6 cfu/mL。
9. The method according to claim 5, wherein the fermentation time in S4 is 4 to 7 hours, and the fermentation temperature is 42 to 44 ℃.
10. The method according to claim 5, wherein the fermentation is terminated at S4 when the pH of the yogurt obtained after fermentation is lowered to 4.6, and the yogurt is stored at a temperature of 2-8 ℃.
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