CN107937317B - Lactobacillus delbrueckii subsp bulgaricus, application thereof and yoghourt prepared from lactobacillus delbrueckii subsp bulgaricus - Google Patents
Lactobacillus delbrueckii subsp bulgaricus, application thereof and yoghourt prepared from lactobacillus delbrueckii subsp bulgaricus Download PDFInfo
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Abstract
The invention belongs to the technical field of dairy products, and particularly relates to lactobacillus delbrueckii subsp bulgaricus, application thereof and yoghourt prepared from the lactobacillus delbrueckii subsp bulgaricus. The strain is Lactobacillus delbrueckii subsp. bulgaricus LB-81, and is preserved in China general microbiological culture collection center with the preservation number of CGMCC No 12958. The lactobacillus delbrueckii subsp bulgaricus is matched with the streptococcus thermophilus for yoghurt fermentation, so that more beneficial bacilli can be provided, the post-acidification is well controlled, and more active bacilli are provided while the refreshing and delicious flavor is kept; the prepared yoghourt is uniform milk white or yellowish, the fermented fragrance is thick and mellow, the tissue is fine and uniform, the acidity is more than 70 DEG T when leaving a factory, the number of viable lactic acid bacteria is 2-7 multiplied by 108cfu/g, and the clearance rate after being stored for 10 days at the temperature of 2-8 ℃ is less than or equal to 1.5%.
Description
Technical Field
The invention belongs to the technical field of dairy products, and particularly relates to lactobacillus delbrueckii subsp bulgaricus, application thereof and yoghourt prepared from the lactobacillus delbrueckii subsp bulgaricus.
Background
In modern society, milk and dairy products have become important components of our daily food, and they have many aspects such as nutritional composition, basic food, leisure food and functional health, which cannot be replaced by other foods, and many countries and regions in the world history use fermented dairy products as the main part of their diets. The yoghourt is a fermented dairy product which is loved by people of various countries, and the sale amount of the yoghourt in 2016 is increased by 24.2% compared with that in 2015. The abundant nutritive value of the yoghourt is acknowledged by consumers, and the yoghourt mainly has the effects of supplementing protein, supplementing calcium element, resisting bacteria, relieving lactose intolerance, improving constipation, improving immunity, resisting cancer and the like.
According to the Chinese regulations, traditional yoghurt shall be called yoghurt. Legislation states that: the yogurt (yoghurt) is a product prepared by taking raw cow (sheep) milk or milk powder as a raw material, sterilizing, inoculating streptococcus thermophilus and lactobacillus delbrueckii subsp bulgaricus (lactobacillus bulgaricus) and fermenting. The yogurt contains only streptococcus thermophilus and lactobacillus bulgaricus, wherein the lactobacillus bulgaricus is the main strain for generating flavor substances. According to other researches, the lactobacillus delbrueckii subsp bulgaricus not only endows the yogurt with special aromatic smell, but also has physiological functions of resisting oxidation, resisting tumors, reducing cholesterol, reducing blood pressure, adjusting intestinal flora, inhibiting bacteria, resisting cancers and the like.
At present, various commercial microbial agents used for producing the yoghourt pay attention to the texture and post-acid control of the yoghourt, the corresponding fragrance of the yoghourt lacks of various changes, and the homogenization is serious. The proportion of the yoghourt strains in the common microbial inoculum at home and abroad is Lactobacillus delbrueckii subspecies Bulgaria: the ratio of streptococcus thermophilus is 1:3-1: 10. Although the product produced by the method can obtain better post-acid control by adding a small amount of lactobacillus delbrueckii subsp bulgaricus with strong acid production capability, the special flavor of the lactobacillus delbrueckii subsp bulgaricus is difficult to obtain, and abnormal fermentation and aroma production are caused by excessively high addition amount. The survival number of the strain with more health efficacy on human body is limited by less bacillus adding amount, so that consumers cannot take more lactobacillus bulgaricus through yoghourt to obtain health effect.
Patent discovery of retrieval of yogurt fermentation using cow milk: the strain used in the patent of the frozen yoghurt and the preparation method thereof is 1: 1.5-3 of lactobacillus bulgaricus and streptococcus thermophilus; in the patent 'a preparation method of a direct-vat-set yoghurt starter' the freeze-dried powder of streptococcus thermophilus and lactobacillus bulgaricus is mixed according to the proportion of 1: 10-1: 100; in the patent of the kelp dietary fiber yoghourt and the production method thereof, the adding proportion of streptococcus thermophilus to lactobacillus bulgaricus is 1:1. Only in the patent of 'a production method of collagen buffalo yoghurt' using buffalo milk as a raw material, the adding proportion of streptococcus thermophilus and lactobacillus bulgaricus reaches 1: 2.3-2.5, but the nutrition (such as various proteins) content and proportion of buffalo milk and milk are greatly different, and the milk serving as the raw material has large fermentation difference compared with milk, and is not suitable for being used as the reference of the patent of the invention.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide lactobacillus delbrueckii subsp bulgaricus which can provide more beneficial bacilli for people who eat the lactobacillus delbrueckii subsp bulgaricus, and also provides application of the lactobacillus delbrueckii subsp bulgaricus in yoghourt fermentation and yoghourt prepared from the lactobacillus delbrueckii subsp bulgaricus, which can better control post-acidification, keep fresh and delicious flavor of products and provide more active bacilli to meet the health requirements of consumers.
The Lactobacillus delbrueckii subsp. bulgaricus strain is Lactobacillus delbrueckii subsp. bulgaricus LB-81, is preserved in China general microbiological culture collection center with the preservation number of CGMCC No 12958.
The Culture is preserved in China General Microbiological Culture Collection Center (CGMCC) of the International Collection of microorganisms designated by the Budapest treaty at 9/12/2016. The address of the depository: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
The Lactobacillus delbrueckii subspecies bulgaricus LB-81 is a gram-positive bacterium, the nucleotide sequence of the bacterium is shown as seq. ID No.1, the bacterium is in a medium-short rod shape, and the size of the thallus of a vegetative cell is (0.6-0.8) Mumx (2.0-4.0) Mum. LB-81 forms a round colony with smooth surface, yellow white color and regular edge on the MRS plate.
The lactobacillus delbrueckii subspecies bulgaricus is applied to yogurt fermentation.
Preferably, lactobacillus delbrueckii subsp bulgaricus is combined with streptococcus thermophilus for yoghurt fermentation. The ratio of Lactobacillus delbrueckii subspecies bulgaricus to Streptococcus thermophilus is preferably in the range of 3:1 to 1:1.
The yoghourt prepared by adopting the lactobacillus delbrueckii subsp bulgaricus provided by the invention comprises the following raw materials: 750Kg of milk, 850Kg of white granulated sugar, 70 Kg to 120Kg of white granulated sugar and 250DCU of basic strains, wherein the basic strains are: the ratio of the Lactobacillus delbrueckii subspecies bulgaricus to the Streptococcus thermophilus is preferably 3:1-1: 1.
Preferably, 0-5Kg of stabilizer is also included, preferably 1-5Kg, including but not limited to: one or more of sodium carboxymethylcellulose, xanthan gum, pectin or modified starch.
As a preferred technical scheme, the production of the product adopts a secondary homogenization process, namely, after the white granulated sugar and the fresh milk are mixed and proportioned, the mixture is homogenized, sterilized, inoculated and fermented to the end point, and then cooled to below 20 ℃; mixing the mixed solution of white sugar and stabilizer, stirring, sterilizing, cooling to below 20 deg.C, mixing the yogurt base material and the mixed solution of stabilizer and sugar water at fermentation end, homogenizing at 180-200bar, bottling, and packaging.
The product of the invention contains more than or equal to 2.3 percent of protein, more than or equal to 2.5 percent of fat and 10 percent of viable bacteria8Per 100g, is a yogurt product rich in high-activity Lactobacillus bulgaricus.
As a preferred technical scheme, the preparation method of the live bacteria yoghourt comprises the step of fermenting the yoghourt according to a conventional method to obtain the yoghourt, wherein after the fermentation of the yoghourt is finished, sugar accounting for 0.5-4% of the weight of the yoghourt is added, and then secondary homogenization is carried out, and a stabilizer accounting for 0.1-0.8% of the weight of the yoghourt can be added at the same time or not added.
The addition conditions of the stabilizer and the sugar are as follows: and (3) cooling the yogurt to below 20 ℃ after the yogurt reaches the fermentation end point, and uniformly mixing the sugar and the stabilizer.
The conditions of the secondary homogenization are as follows: homogenizing under 18-20Mpa at 65-70 deg.C.
Before the secondary homogenization, the steps of cylinder turning and cooling can also be carried out, and the specific method comprises the following steps: and (3) opening ice water after the acidity reaches the requirement, inching the stirrer for a certain time, preferably 30s, turning the cylinder to the buffer tank, and controlling the temperature to be 18-20 ℃.
As a preferred technical scheme, the preparation method of the yoghourt comprises the following steps:
(1) adding white granulated sugar into fresh milk, mixing, stirring, homogenizing, sterilizing, inoculating, fermenting, and cooling after fermentation reaches the end point;
(2) uniformly mixing the white granulated sugar and the stabilizer, adding the mixture into water, sterilizing and cooling;
(3) and (3) uniformly stirring the yogurt base material with the fermentation end point obtained in the step (1) and the mixed solution of the stabilizer and the sugar water obtained in the step (2), homogenizing, filling and packaging.
The homogenizing and sterilizing in the step (1) are as follows: homogenizing under 18-20Mpa at 65-70 deg.C, and sterilizing at 95-100 deg.C/300 s; the fermentation is carried out at the temperature of 43 +/-2 ℃, and the fermentation end point is as follows: the end point acidity is controlled at 70-80T.
The homogenization in the step (3) is as follows: homogenizing under 18-20Mpa at 65-70 deg.C.
As a preferable technical scheme, the preparation method of the yoghourt comprises the following steps:
(1) detecting raw milk;
(2) standardizing raw milk;
(3) homogenizing and sterilizing;
(4) cooling and then inoculating;
(5) fermenting by a conventional method;
(6) turning over the cylinder and cooling;
(7) preparing sugar and stabilizer solution, uniformly mixing the sugar and the stabilizer, uniformly stirring, sterilizing and cooling;
(8) homogenizing for the second time;
(9) and (6) filling.
The raw milk detection steps are as follows: the 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃;
the raw milk standardization step comprises: milk temperature (45-50 ℃) and circulation time (15min), standardized fat and milk solids-not-fat must meet the specified requirements: fat is more than or equal to 3.1 percent, and non-fat milk solid is more than or equal to 8.1 percent;
the homogenizing and sterilizing steps are as follows: homogenizing at 18-20Mpa/65-70 deg.C, and sterilizing at 95-100 deg.C/300 s;
the cooling and inoculation steps are as follows: cooling the materials to 43-45 deg.C, adding basic strain, mixing for 20min, and turning off the stirrer;
the fermentation steps are as follows: fermenting at 43 + -2 deg.C, and controlling the final acidity at 70-80 deg.C T;
the fermentation is that the starter inoculum accounts for 2-5% of the volume ratio of the raw milk, the fermentation is carried out for 3-6 h at 40 +/-5 ℃, and the final acidity is 70-80 DEG T;
the preparation steps of the sugar and stabilizer solution are as follows: mixing sugar and stabilizer, slowly adding into hot water of 65-70 deg.C, mixing and stirring. Sterilizing at 95-100 deg.C/300 s, and cooling to below 20 deg.C;
the filling step is followed by warehousing and refrigeration, and the method comprises the following steps: cooling to 10 deg.C within 1h, and refrigerating at 2-6 deg.C for 24 h.
More preferably, the preparation method of the yoghurt comprises the following steps:
(1) detecting raw milk;
(2) standardizing raw milk;
(3) yogurt base material ingredients: adding 40Kg of white granulated sugar into 750-850Kg of fresh milk heated to 65-70 ℃, mixing and stirring, and cooling to below 10 ℃;
(4) homogenizing and sterilizing: homogenizing under 18-20Mpa at 65-70 deg.C, and sterilizing at 95-100 deg.C/300 s;
(5) cooling and inoculating: cooling to 43-45 deg.C, adding basic seed, and mixing;
(6) fermentation: fermenting at 43 +/-2 ℃, and controlling the end-point acidity to be 70-80 DEG T:
(7) and (3) cooling: the ice water was turned on and the temperature was controlled below 20 ℃:
(8) preparing a sugar and stabilizer solution: adding 20-50Kg of sugar and stabilizer after mixing uniformly into 65-70 deg.C hot water, or adding 20-50Kg of sugar into 65-70 deg.C hot water, mixing and stirring uniformly, sterilizing at 95-100 deg.C/300 s, cooling to below 20 deg.C after sterilization to obtain yogurt base material;
(9) and (3) secondary homogenization: uniformly mixing the yogurt base material obtained in the step 7 and cooled to below 20 ℃ after reaching the fermentation end point and the mixed solution of the sugar and the stabilizer obtained in the step 8 and cooled to below 20 ℃, and homogenizing under the condition of 18-20Mpa/65-70 ℃;
(10) and (6) filling and packaging.
The amount of water in the step (8) is based on the capability of dissolving the sugar and the stabilizing agent or the sugar and the stabilizing agent are prepared into pasty dry powder-free powder.
And after the packaging is finished, putting the mixture into a warehouse for refrigeration, reducing the temperature to 10 ℃ within 1 hour, and then refrigerating the mixture for 24 hours at the temperature of 2-6 ℃.
In the preparation method of the product, the amount of the sugar added before the yogurt is fermented is the conventional technology, the influence of the amount of the sugar added on the product is not great, and the key points are the amount of the sugar and the amount of the stabilizer added after the fermentation and before the secondary homogenization.
After the product is prepared, taste evaluation is carried out, and 50 people are subjected to scoring evaluation in an anonymous scoring mode. 9 indexes of front aroma, back aroma, natural feeling, thin consistency, sticky mouthfeel, sweetness, acidity, smoothness, refreshing degree and integral feeling are 10 points respectively, the full score is 100 points, the higher the score is, the better the effect is, the statistical analysis is carried out on the tasting result, the questionnaire with the total score of 75 points and above is the qualified questionnaire of mouthfeel, and the quality of the product is verified when the quantity of the qualified questionnaire of mouthfeel exceeds 70%.
More preferably, the preparation method of the yoghurt comprises the following detailed steps:
1. and (5) detecting the raw milk. The 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃. And after the detection is qualified, the next procedure (control point) can be carried out.
2. Standardizing raw milk. When in standardization, the milk temperature (45-50 ℃) and the circulation time (15min) are noticed, and the fat and the milk solid non-fat after standardization must meet the specified requirements;
3. and (4) yogurt base material ingredients. 40Kg of sugar was slowly added to 850Kg of fresh milk heated to 65-70 deg.C and 750-. Cooling to below 10 deg.C before sterilization. And after the detection is qualified, the next procedure (control point) can be carried out.
4. Homogenizing and sterilizing. Homogenizing under 18-20Mpa at 65-70 deg.C, and sterilizing at 95-100 deg.C/300 s. And after the detection is qualified, the next procedure (control point) can be carried out.
5. Cooling and inoculating. The material was cooled to 43-45 ℃ as per bacillus: adding basic strains (Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus, the same below) at a ratio of 3:1-1:1, mixing for 20min, and turning off the stirrer.
6. And (5) fermenting. Fermenting at 43 + -2 deg.C, and controlling final acidity at 70-80 deg.C T.
7. And (6) cooling. And (3) after the acidity meets the requirement, opening ice water, inching the stirrer for 30s, turning the cylinder to the buffer tank, controlling the temperature to be 18-20 ℃, and carefully regulating the flow.
8. And (3) preparing a sugar and stabilizer solution. After 20-50Kg of sugar and stabilizer are mixed evenly, the mixture is slowly added into hot water with the temperature of 65-70 ℃, the amount of the water is based on the fact that the sugar and the stabilizer can be just dissolved or the sugar and the stabilizer are prepared into pasty dry powder, and the mixture is mixed and stirred evenly. Sterilizing at 95-100 deg.C/300 s, and cooling to below 20 deg.C.
9. And (5) homogenizing for the second time. Cooling to below 20 deg.C after reaching fermentation end point, mixing sugar and stabilizer mixed solution uniformly, and homogenizing at 18-20Mpa/65-70 deg.C.
10. Filling and packaging
11. And (5) warehousing and refrigerating. Cooling to 10 deg.C within 1h, and refrigerating at 2-6 deg.C for 24 h.
12. And (6) checking.
In summary, the invention has the following advantages:
the invention provides a Lactobacillus delbrueckii subspecies bulgaricus which can provide more beneficial bacilli for edible people, and the invention also providesThe maximum adding proportion of the Lactobacillus delbrueckii subsp bulgaricus and the Streptococcus thermophilus can reach 3:1, the proportion of the Lactobacillus delbrueckii subsp bulgaricus in the yoghurt is increased, more beneficial bacilli are provided, the post-acidification can be well controlled, the fresh and delicious flavor of the product is kept, and more active bacilli can be provided to meet the health requirements of consumers. The yogurt prepared by the invention has uniform milky white or yellowish appearance color, inherent taste and smell of the yogurt, strong and mellow fermentation fragrance, fine and uniform tissue, acidity higher than 70 DEG T when the product leaves the factory, and viable lactic acid bacteria number of 2-7 multiplied by 108cfu/g, and the clearance rate of the product after being stored for 10 days at the temperature of 2-8 ℃ is less than or equal to 1.5 percent.
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1
1. And (5) detecting the raw milk. The 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃. And after the detection is qualified, the next procedure (control point) can be carried out.
2. Standardizing raw milk. Milk temperature (50 ℃) and circulation time (15min) are standardized, and fat and non-fat milk solids must meet the specified requirements after standardization;
3. and (4) yogurt base material ingredients. 40Kg of sugar was slowly added to 850Kg of fresh milk heated to 67 deg.C, mixed and stirred for 10 min. Cooling to below 10 deg.C before sterilization. And after the detection is qualified, the next procedure (control point) can be carried out.
4. Homogenizing and sterilizing. Homogenizing under 20Mpa/65 deg.C, and sterilizing at 95 deg.C/300 s. And after the detection is qualified, the next procedure (control point) can be carried out.
5. Cooling and inoculating. The batch was cooled to 45 ℃ as follows: adding basic strain at a ratio of 3:1, mixing for 20min, and turning off the stirrer.
6. And (5) fermenting. Fermenting at 45 deg.C, and controlling final acidity to be more than 70 deg.C T.
7. And (6) cooling. And (4) after the acidity meets the requirement, opening ice water, inching the stirrer for 30s, turning the cylinder to the buffer tank, controlling the temperature at 20 ℃, and carefully regulating the flow.
8. And (5) preparing a sugar solution. 20Kg of sugar was slowly added to hot water at 70 ℃ and mixed well. Sterilizing at 95 deg.C/300 s, and cooling to below 20 deg.C.
9. And (5) homogenizing for the second time. Mixing yogurt base material cooled to below 20 deg.C after reaching fermentation end point with sugar cooled to below 20 deg.C, and homogenizing at 20Mpa/65 deg.C.
10. And (6) filling and packaging.
11. And (5) warehousing and refrigerating. The temperature is reduced to 10 ℃ within 1h, and then the mixture is refrigerated for 24h at the temperature of 2 ℃.
12. The test result is as follows: the acidity is 78 DEG T, the quantity of questionnaires qualified in taste is 84%, and the product ingredients are not separated or float upwards.
Example 2
1. And (5) detecting the raw milk. The 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃. And after the detection is qualified, the next procedure (control point) can be carried out.
2. Standardizing raw milk. When in standardization, the milk temperature (45 ℃) and the circulation time (15min) are noticed, and after standardization, the fat and the non-fat milk solid must meet the specified requirements;
3. and (4) yogurt base material ingredients. 40Kg of sugar was slowly added to 810Kg of fresh milk heated to 70 ℃ and mixed and stirred for 10 min. Cooling to below 10 deg.C before sterilization. And after the detection is qualified, the next procedure (control point) can be carried out.
4. Homogenizing and sterilizing. Homogenizing under 20Mpa/65 deg.C, and sterilizing at 95 deg.C/300 s. And after the detection is qualified, the next procedure (control point) can be carried out.
5. Cooling and inoculating. The batch was cooled to 43 ℃ as follows: adding basic strain at a ratio of 3:1, mixing for 20min, and turning off the stirrer.
6. And (5) fermenting. Fermenting at 41 deg.C, and controlling final acidity to be more than 70 deg.C T.
7. And (6) cooling. And (4) after the acidity meets the requirement, opening ice water, inching the stirrer for 30s, turning the cylinder to the buffer tank, controlling the temperature to be 18 ℃, and carefully regulating the flow.
8. And (5) preparing a sugar solution. 50Kg of sugar was slowly added to hot water at 65-70 deg.C, and mixed and stirred uniformly. Sterilizing at 100 deg.C/300 s, and cooling to below 20 deg.C.
9. And (5) homogenizing for the second time. Mixing yogurt base material cooled to below 20 deg.C after reaching fermentation end point with sugar solution cooled to below 20 deg.C, and homogenizing at 18Mpa/70 deg.C.
10. And (6) filling and packaging.
11. And (5) warehousing and refrigerating. The temperature is reduced to 10 ℃ within 1h, and then the mixture is refrigerated for 24h at 6 ℃.
12. The test result is as follows: the acidity is 76 degrees T, the quantity of questionnaires qualified in taste is 90 percent, and the product ingredients are not separated or floated.
Example 3
1. And (5) detecting the raw milk. The 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃. And after the detection is qualified, the next procedure (control point) can be carried out.
2. Standardizing raw milk. When in standardization, the milk temperature (48 ℃) and the circulation time (15min) are noticed, and after standardization, the fat and the non-fat milk solid must meet the specified requirements;
3. and (4) yogurt base material ingredients. 40Kg of sugar was slowly added to 850Kg of fresh milk heated to 65 deg.C, mixed and stirred for 10 min. Cooling to below 10 deg.C before sterilization. And after the detection is qualified, the next procedure (control point) can be carried out.
4. Homogenizing and sterilizing. Homogenizing at 20Mpa/65 deg.C, and sterilizing at 98 deg.C/300 s. And after the detection is qualified, the next procedure (control point) can be carried out.
5. Cooling and inoculating. The batch was cooled to 44 ℃ as follows: adding basic strain at a ratio of 1:1, mixing for 20min, and turning off the stirrer.
6. And (5) fermenting. Fermenting at 43 deg.C, and controlling final acidity to be more than 70 deg.C T.
7. And (6) cooling. And (4) after the acidity meets the requirement, opening ice water, inching the stirrer for 30s, turning the cylinder to the buffer tank, controlling the temperature to be 19 ℃, and carefully regulating the flow.
8. And (5) preparing a sugar solution. 20Kg of sugar was slowly added to hot water at 68 ℃ and mixed well. Sterilizing at 95 deg.C/300 s, and cooling to below 20 deg.C.
9. And (5) homogenizing for the second time. Mixing yogurt base material cooled to below 20 deg.C after reaching fermentation end point with sugar solution cooled to below 20 deg.C, and homogenizing at 20Mpa/65 deg.C.
10. And (6) filling and packaging.
11. And (5) warehousing and refrigerating. The temperature is reduced to 10 ℃ within 1h, and then the mixture is refrigerated for 24h at 4 ℃.
12. The test result is as follows: the acidity is 73 degrees T, the quantity of questionnaires qualified in taste is 92 percent, and the product ingredients are not separated or float upwards.
Example 4
1. And (5) detecting the raw milk. The 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃. And after the detection is qualified, the next procedure (control point) can be carried out.
2. Standardizing raw milk. Milk temperature (45 ℃) and circulation time (15min) were noted for standardization, and fat and milk solids-not-fat had to meet the specifications after standardization.
3. And (4) yogurt base material ingredients. 40Kg of sugar was slowly added to 810Kg of fresh milk heated to 68 ℃ and mixed and stirred for 10 min. Cooling to below 10 deg.C before sterilization. And after the detection is qualified, the next procedure (control point) can be carried out.
4. Homogenizing and sterilizing. Homogenizing under 20Mpa/65 deg.C, and sterilizing at 95 deg.C/300 s. And after the detection is qualified, the next procedure (control point) can be carried out.
5. Cooling and inoculating. The batch was cooled to 43 ℃ as follows: adding basic strain at a ratio of 1:1, mixing for 20min, and turning off the stirrer.
6. And (5) fermenting. Fermenting at 42 deg.C, and controlling final acidity to be more than 70 deg.C T.
7. And (6) cooling. And (4) after the acidity meets the requirement, opening ice water, inching the stirrer for 30s, turning the cylinder to the buffer tank, controlling the temperature at 20 ℃, and carefully regulating the flow.
8. And (3) preparing a sugar and stabilizer solution. 50Kg of sugar was slowly added to hot water at 65 ℃ and mixed well. Sterilizing at 95 deg.C/300 s, and cooling to below 20 deg.C.
9. And (5) homogenizing for the second time. Mixing yogurt base material cooled to below 20 deg.C after reaching fermentation end point with sugar solution cooled to below 20 deg.C, and homogenizing at 19Mpa/65 deg.C.
10. And (6) filling and packaging.
11. And (5) warehousing and refrigerating. The temperature is reduced to 10 ℃ within 1h, and then the mixture is refrigerated for 24h at the temperature of 2 ℃.
12. The test result is as follows: the acidity is 77 DEG T, the quantity of questionnaires qualified in taste is 88 percent, and the product ingredients are not separated or float upwards.
Example 5
1. And (5) detecting the raw milk. The 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃. And after the detection is qualified, the next procedure (control point) can be carried out.
2. Standardizing raw milk. Milk temperature (45-50 ℃) and circulation time (15min) are noted during standardization, and after standardization, fat and milk solids-not-fat must meet the specified requirements.
3. And (4) yogurt base material ingredients. 40Kg of sugar was slowly added to 750Kg of fresh milk heated to 70 ℃ and mixed and stirred for 10 min. Cooling to below 10 deg.C before sterilization. And after the detection is qualified, the next procedure (control point) can be carried out.
4. Homogenizing and sterilizing. Homogenizing under 20Mpa/65 deg.C, and sterilizing at 95 deg.C/300 s. And after the detection is qualified, the next procedure (control point) can be carried out.
5. Cooling and inoculating. Cooling the material to 43 deg.C, adding basic strain at a ratio of 3:1, mixing for 20min, and turning off the stirrer.
6. And (5) fermenting. Fermenting at 43 deg.C, and controlling final acidity to be more than 70 deg.C T.
7. And (6) cooling. And (4) after the acidity meets the requirement, opening ice water, inching the stirrer for 30s, turning the cylinder to the buffer tank, controlling the temperature to be 18 ℃, and carefully regulating the flow.
8. And (3) preparing a sugar and stabilizer solution. After 25Kg of white granulated sugar and 5Kg of stabilizer are mixed uniformly, the mixture is slowly added into hot water with the temperature of 65 ℃, and the mixture is mixed and stirred uniformly. Sterilizing at 95 deg.C/300 s, and cooling to below 20 deg.C.
9. And (5) homogenizing for the second time. Cooling the yogurt base material to below 20 deg.C after reaching the fermentation end point, cooling the sugar to below 20 deg.C, mixing with the stabilizer mixture, and homogenizing at 20Mpa/65 deg.C.
10. And (6) filling and packaging.
11. And (5) warehousing and refrigerating. Cooling to 10 deg.C within 1h, and refrigerating at 2-6 deg.C for 24 h.
12. The test result is as follows: the acidity is 76 degrees T, the quantity of questionnaires qualified in taste is 88 percent, and the product ingredients are not separated or float upwards.
Example 6
1. And (5) detecting the raw milk. The 72% alcohol protein is stable, the acidity is less than or equal to 18 DEG T, and the boiling is normal; detecting fat, protein, non-fat milk solids and antibiotic residues; the milk temperature is less than or equal to 4 ℃. And after the detection is qualified, the next procedure (control point) can be carried out.
2. Standardizing raw milk. Milk temperature (45-50 ℃) and circulation time (15min) are noted during standardization, and after standardization, fat and milk solids-not-fat must meet the specified requirements.
3. And (4) yogurt base material ingredients. 40Kg of sugar was slowly added to 750Kg of fresh milk heated to 65 ℃ and mixed and stirred for 10 min. Cooling to below 10 deg.C before sterilization. And after the detection is qualified, the next procedure (control point) can be carried out.
4. Homogenizing and sterilizing. Homogenizing under 20Mpa/65 deg.C, and sterilizing at 95 deg.C/300 s. And after the detection is qualified, the next procedure (control point) can be carried out.
5. Cooling and inoculating. Cooling the material to 43 deg.C, adding basic strain at a ratio of 1:1, mixing for 20min, and turning off the stirrer.
6. And (5) fermenting. Fermenting at 43 deg.C, and controlling final acidity to be more than 70 deg.C T.
7. And (6) cooling. And (4) after the acidity meets the requirement, opening ice water, inching the stirrer for 30s, turning the cylinder to the buffer tank, controlling the temperature to be 18 ℃, and carefully regulating the flow.
8. And (3) preparing a sugar and stabilizer solution. After 25Kg of white granulated sugar and 5Kg of stabilizer are mixed uniformly, the mixture is slowly added into hot water with the temperature of 65 ℃, and the mixture is mixed and stirred uniformly. Sterilizing at 95 deg.C/300 s, and cooling to below 20 deg.C.
9. And (5) homogenizing for the second time. Cooling the yogurt base material to below 20 deg.C after reaching the fermentation end point, cooling the sugar to below 20 deg.C, mixing with the stabilizer mixture, and homogenizing at 20Mpa/65 deg.C.
10. And (6) filling and packaging.
11. And (5) warehousing and refrigerating. Cooling to 10 deg.C within 1h, and refrigerating at 2-6 deg.C for 24 h.
12. The test result is as follows: the acidity is 78 degrees T, the quantity of questionnaires qualified in taste is 80 percent, and the product ingredients are not separated or float upwards.
Sequence listing
<110> Shandong De Yi milk industry Co., Ltd
<120> Lactobacillus delbrueckii subsp bulgaricus, application thereof and yoghourt prepared from same
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1473
<212> DNA
<213> Lactobacillus delbrueckii subsp. bulgaricus (Lactobacillus delbrueckii subsp. bulgaricus)
<400> 1
actcctataa aggttatccc accgactttg ggcattgcag acttccatgg tgtgacgggc 60
ggtgtgtaca aggcccggga acgtattcac cgcggcgtgc tgatccgcga ttactagcga 120
ttccagcttc gtgcaggcga gttgcagcct gcagtccgaa ctgagaacag ctttaagaga 180
tccgcttacc ctcgcgggtt cgcttctcgt tgtactgccc attgtagcac gtgtgtagcc 240
caggtcataa ggggcatgat gacttgacgt catccccacc ttcctccggt ttgtcaccgg 300
cagtctcttt agagtgccca acttaatgat ggcaactaaa gacaagggtt gcgctcgttg 360
cgggacttaa cccaacatct cacgacacga gctgacgaca gccatgcacc acctgtctct 420
gcgtccccga agggaaccac ctatctctag gtgtagcaca ggatgtcaag acctggtaag 480
gttcttcgcg ttgcttcgaa ttaaaccaca tgctccaccg cttgtgcggg cccccgtcaa 540
ttcctttgag tttcaacctt gcggtcgtac tccccaggcg gagcgcttaa tgcgtttgct 600
gcggcactga gaaccggaaa gtccccaaca cctagcgctc atcgtttacg gcatggacta 660
ccagggtatc taatcctgtt cgctacccat gctttcgagc ctcagcgtca gttgcagacc 720
agagagccgc cttcgccact ggtgttcttc catatatcta cgcattccac cgctacacat 780
ggaattccac tctcctcttc tgcactcaag aatgacagtt tccgatgcag ttccacggtt 840
gagccgtggg ctttcacatc agacttatca ttccgcctgc gctcgcttta cgcccaataa 900
atccggacaa cgcttgccac ctacgtatta ccgcggctgc tggcacgtag ttagccgtga 960
ctttctggtt gattaccgtc aaataaagac cagttactgc ctctatcctt cttcaccaac 1020
aacagagctt tacgatccga aaaccttctt cactcacgcg gcgttgctcc atcagacttg 1080
cgtccattgt ggaagattcc ctactgctgc ctcccgtagg agtttgggcc gtgtctcagt 1140
cccaatgtgg ccgatcagtc tctcaactcg gctacgcatc attgccttgg taggccttta 1200
ccccaccaac tagctaatgc gccgcgggct catcctaaag tgacagcttg cgccgccttt 1260
caaacttgaa tcatgcgatt catgttgtta tccggtatta gcacctgttt ccaagtggta 1320
tcccagtctt tagggcagat tgcccacgtg ttactcaccc atccgccgct agcgtccaac 1380
aaatcacccc gaaggaatct ttgaattcag ctcgctcgac ttgcatgtat taggcacgcc 1440
gccagcgttc gtcctgagcc aggatcaaac tct 1473
Claims (2)
1. The application of lactobacillus delbrueckii subspecies bulgaricus is characterized in that: the Lactobacillus delbrueckii subsp. bulgaricus LB-81 is preserved in China general microbiological culture collection center with the preservation number of CGMCC No 12958, and the Lactobacillus delbrueckii subsp. bulgaricus and the streptococcus thermophilus are matched for yoghurt fermentation, wherein the ratio of the Lactobacillus delbrueckii subsp. bulgaricus to the streptococcus thermophilus is 3: 1.
2. The use of Lactobacillus delbrueckii subsp bulgaricus according to claim 1, characterized in that: the preparation method of the yoghourt comprises the following steps:
(1) detecting raw milk;
(2) standardizing raw milk;
(3) yogurt base material ingredients: adding 40Kg of white granulated sugar into 750-850Kg of fresh milk heated to 65-70 ℃, mixing and stirring, and cooling to below 10 ℃;
(4) homogenizing and sterilizing: homogenizing under 18-20Mpa at 65-70 deg.C, and sterilizing at 95-100 deg.C/300 s;
(5) cooling and inoculating: cooling to 43-45 deg.C, adding basic seed, and mixing;
(6) fermentation: fermenting at 43 +/-2 ℃, and controlling the end-point acidity to be 70-80 DEG T:
(7) and (3) cooling: the ice water was turned on and the temperature was controlled below 20 ℃:
(8) preparing a sugar and stabilizer solution: adding 20-50Kg of sugar and stabilizer after mixing uniformly into 65-70 deg.C hot water, or adding 20-50Kg of sugar into 65-70 deg.C hot water, mixing and stirring uniformly, sterilizing at 95-100 deg.C/300 s, cooling to below 20 deg.C after sterilization to obtain yogurt base material;
(9) and (3) secondary homogenization: uniformly mixing the yogurt base material obtained in the step 7 and cooled to below 20 ℃ after reaching the fermentation end point and the mixed solution of the sugar and the stabilizer obtained in the step 8 and cooled to below 20 ℃, and homogenizing under the condition of 18-20Mpa/65-70 ℃;
(10) and (6) filling and packaging.
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| CN110100896A (en) * | 2019-06-18 | 2019-08-09 | 甘南藏族自治州燎原乳业有限责任公司 | A kind of yak milk flavor yoghourt and preparation method thereof |
| RU2731738C1 (en) * | 2019-11-11 | 2020-09-08 | Федеральное бюджетное учреждение науки "Московский научно-исследовательский институт эпидемиологии и микробиологии им. Г.Н. Габричевского" Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека | Bacterial strain lactobacillus delbrueckii subsp_ bulgaricus k 1903, used as starter of direct application for preparation of fermented milk products |
| CN114532400B (en) * | 2021-12-22 | 2023-09-01 | 山东得益乳业股份有限公司 | Streptococcus thermophilus and yoghourt prepared by using same |
| CN114391577A (en) * | 2021-12-30 | 2022-04-26 | 山东得益乳业股份有限公司 | High-calcium flavored fermented milk and preparation method thereof |
| CN114854623B (en) * | 2022-03-15 | 2023-09-29 | 泸州老窖股份有限公司 | Lactobacillus delbrueckii subspecies bulgaricus, microbial inoculum containing lactobacillus delbrueckii subspecies bulgaricus and application of lactobacillus delbrueckii subspecies bulgaricus |
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