CN113088474A - Microbial combined leavening agent and preparation method and application thereof - Google Patents
Microbial combined leavening agent and preparation method and application thereof Download PDFInfo
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- CN113088474A CN113088474A CN202110523041.9A CN202110523041A CN113088474A CN 113088474 A CN113088474 A CN 113088474A CN 202110523041 A CN202110523041 A CN 202110523041A CN 113088474 A CN113088474 A CN 113088474A
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- streptococcus thermophilus
- microbial
- lactobacillus
- starter
- lactobacillus paracasei
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- 241000194020 Streptococcus thermophilus Species 0.000 claims abstract description 67
- 240000002605 Lactobacillus helveticus Species 0.000 claims abstract description 65
- 238000000855 fermentation Methods 0.000 claims abstract description 63
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Images
Classifications
-
- 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.
-
- 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
- A23C9/1238—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using specific L. bulgaricus or S. thermophilus microorganisms; using entrapped or encapsulated yoghurt bacteria; Physical or chemical treatment of L. bulgaricus or S. thermophilus cultures; Fermentation only with L. bulgaricus or only with S. thermophilus
-
- 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
- A23V2400/147—Helveticus
-
- 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
- A23V2400/165—Paracasei
-
- 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
Abstract
The invention relates to the technical field of microbial fermentation, and particularly discloses a microbial combined leavening agent as well as a preparation method and application thereof. The microbial combined leaven comprises lactobacillus helveticus L551, lactobacillus paracasei L578 and streptococcus thermophilus S56. The microbial combined starter can be used as a direct-vat-set yoghurt starter. The microbial combined starter provided by the invention has the advantages of a fermentation period and good flavor of a fermentation product, the fermentation product can produce gamma-aminobutyric acid at a high yield, and volatile short-chain fatty acid and aroma substances with high content are produced in the fermented milk, so that the obtained fermented milk has strong cheese flavor of acetic acid flavor.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to a microbial combined leavening agent and a preparation method and application thereof.
Background
The yoghourt is prepared by taking milk as a raw material, adding beneficial bacteria into the milk after pasteurization and fermenting, is a folk beverage which is rich in nutrition and easy to digest, can enhance the immune function of a human body and reduce the level of serum cholesterol, even under the condition of not using any medicine, about 240 g of yoghourt is used per meal, and the cholesterol is reduced after one week. In addition, the normal drinking of the yoghourt can promote the intestinal movement, soften glycolysis colon contents, increase the excrement excretion, prevent the occurrence of constipation and is beneficial to the prevention of colon cancer. Because of the above advantages, the yogurt industry is rapidly developing and favored by people.
In the process of yoghurt fermentation preparation, a yoghurt starter becomes a key factor influencing the quality of yoghurt, and is the basis and main reason for producing fragrance and acid of yoghurt products. Therefore, the quality and flavor of the yogurt mainly depend on the quality, type and vitality of the yogurt starter. The yoghurt starter also becomes one of the traditional Chinese medicine factors restricting the development of the yoghurt industry.
A direct-vat set yoghurt starter is a starter without edible essence. The edible essence is formed by combining various organic compounds and is added into a leavening agent, so that the fermentation activity and the storage stability of the strain can be reduced. However, if essence is not added, the flavor and mouthfeel of the yogurt product are affected. Therefore, the existing yoghourt is prepared by adjusting the essence before fermentation or adding the essence after fermentation to make the yoghourt have the required flavor. This results in a complicated fermentation process of yogurt and increased production costs.
Disclosure of Invention
Aiming at the problems of the existing yoghurt starter, the invention provides a microbial combined starter, a preparation method and application thereof.
In order to achieve the purpose of the invention, the embodiment of the invention adopts the following technical scheme:
a microbial composite starter, comprising the following strains:
lactobacillus helveticus L551, lactobacillus paracasei L578, and streptococcus thermophilus S56;
the strain preservation number of the Lactobacillus helveticus L551 is CGMCC No. 15604; lactobacillus helveticus L551 is a strain disclosed in patent CN111685328A and in patent application of Naqu 4580 probiotics and preparations thereof in foods for regulating emotion, and is preserved in the China general microbiological culture Collection center.
The strain preservation number of the lactobacillus paracasei L578 is CGMCC No. 15603; lactobacillus paracasei L578 strain disclosed in patent CN111685328A and patent application of Naqu 4580 probiotics and preparations thereof in foods for regulating emotion is preserved in the China general microbiological culture Collection center of the Committee for culture Collection of microorganisms.
The strain preservation number of the streptococcus thermophilus S56 is CGMCC No.22066, and the Latin name is as follows: streptococcus thermophilus is preserved in China general microbiological culture Collection center at 25.03.2021, and the preservation addresses are as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
Compared with the prior art, the compound of the lactobacillus helveticus, the lactobacillus paracasei and the streptococcus thermophilus in the microbial combined starter provided by the invention has the advantages of a fermentation period and good flavor of a fermentation product. The fermented milk can produce gamma-aminobutyric acid with high yield, and volatile short-chain fatty acids (represented by acetic acid and a certain amount of caproic acid and caprylic acid) and aroma substances (butanedione and acetoin) with high content are produced in the fermented milk, so that the obtained fermented milk has a strong cheese flavor of the acetic acid flavor, and the fermented milk can keep the strong aroma for a long time without adding any essence.
Preferably, in the microbial combined starter, the ratio of the viable count of the Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56 is 350-450:200-400: 20-50.
The compounding of the ratio of the thalli can realize the synergistic growth of the thalli and further shorten the fermentation period of the thalli.
Preferably, the invention also provides a preparation method of the microbial combined leaven, which comprises the following steps:
a. activating the Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56, and then respectively adding the activated Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56 into a liquid culture medium for fermentation at 37-45 ℃ for 10-12h to obtain Lactobacillus helveticus fermentation liquid, Lactobacillus paracasei fermentation liquid and Streptococcus thermophilus fermentation liquid;
b. respectively centrifuging the lactobacillus helveticus fermentation liquor, the lactobacillus paracasei fermentation liquor and the streptococcus thermophilus fermentation liquor, and separating thalli to obtain lactobacillus helveticus bacterial mud, lactobacillus paracasei bacterial mud and streptococcus thermophilus bacterial mud;
c. respectively adding freeze-drying protective agents into the lactobacillus helveticus bacterial mud, the lactobacillus paracasei bacterial mud and the streptococcus thermophilus bacterial mud, and then carrying out freeze drying to obtain lactobacillus helveticus bacterial powder, lactobacillus paracasei bacterial powder and streptococcus thermophilus bacterial powder;
d. and compounding the lactobacillus helveticus bacterium powder, the lactobacillus paracasei bacterium powder and the streptococcus thermophilus bacterium powder according to the proportion of the viable count of the lactobacillus helveticus, the lactobacillus paracasei and the streptococcus thermophilus in the microbial combined starter to obtain the microbial combined starter.
Compared with the prior art, the preparation method of the microbial combined starter provided by the invention is simple to operate, can be completed by conventional thallus fermentation equipment, is short in fermentation time, and has good thallus activity and high viable bacteria content.
Preferably, in the step a, the liquid culture medium comprises the following components in percentage by mass: glucose: 2.5-3.5%, yeast extract powder: 1-2%, soy peptone: 0.8-1.2%, disodium hydrogen phosphate: 0.13-0.17%, sodium dihydrogen phosphate: 0.2-0.3%, magnesium sulfate: 0.01-0.03%, ascorbic acid: 0.2-0.4%, tween 80: 0.06-0.08% and the balance of sterile water.
Preferably, the freeze-drying protective agent comprises the following components in percentage by mass: 8-15% of skim milk, 5-12% of trehalose, 0.5-5% of monosodium glutamate, 1-5% of sucrose, 0.5-3% of glycerol, 0.1-3% of sodium ascorbate and the balance of sterile water; the addition amount of the freeze-drying protective agent is 85-200% of the mass of the bacterial sludge. More preferably, the freeze-drying protective agent comprises the following components in percentage by mass: 10% of skim milk, 8% of trehalose, 3% of monosodium glutamate, 2% of sucrose, 1% of glycerol, 1% of sodium ascorbate and the balance of sterile water.
The invention also provides application of the microbial combined starter in serving as a direct vat set yogurt starter.
The microbial combined starter can be used as a direct vat set yoghurt starter to directly obtain a yoghurt product with special flavor and strong fragrance, and the addition of essence is avoided, so that the preparation steps and the cost of yoghurt are simplified.
The invention also provides a preparation method of the yoghourt, which comprises the following steps:
s1, heating fresh milk to 50-55 ℃, adding white granulated sugar, whey powder and sodium glutamate, stirring for dissolving, and heating to 55-60 ℃ to obtain milk liquid;
s2, homogenizing and pasteurizing the milk, cooling to 37-45 ℃, adding the microbial combined leaven, fermenting at 37-45 ℃, and stirring, demulsifying, cooling and refining when the pH of the fermented liquid is less than or equal to 4 and the acidity is more than or equal to 110 DEG T to obtain the yoghourt.
The preparation method of the yoghourt provided by the invention is simple to operate and high in fermentation speed, and the prepared yoghourt has a special cheese acetate fragrance flavor and a good taste.
Preferably, in the step S1, the mass ratio of the fresh milk, the white granulated sugar, the whey powder and the sodium glutamate is 850-110: 2-10: 0.5-2.
Preferably, in step S2, the mass ratio of the microbial composite starter to the fresh milk is 0.01-0.08: 850-920.
Preferably, in step S2, the homogenizing temperature is 50-75 deg.C and the pressure is 5-20 MPa.
Preferably, in step S2, the temperature of pasteurization is 94-96 deg.C for 300-350S.
The invention also provides the yoghourt which is prepared by the preparation method of the yoghourt.
Drawings
FIG. 1 is a spectrum of yogurt produced in example 1 detected by GC-MS detection in test example 1 of the present invention;
FIG. 2 is a standard graph showing the acetic acid content obtained in test example 1 of the present invention;
FIG. 3 is a chromatogram of an acetic acid standard in test example 1 of the present invention;
FIG. 4 is a chromatogram for measuring the acetic acid content in the yogurt obtained in example 1 in test example 1 of the present invention;
FIG. 5 is a chromatogram for measuring the acetic acid content in a commercially available yogurt 1 in test example 1 of the present invention;
FIG. 6 is a chromatogram for measuring the acetic acid content in a commercially available yogurt 2 in test example 1 of the present invention;
FIG. 7 is a chromatogram for measuring the content of gamma-aminobutyric acid in the yogurt obtained in example 1 of the present invention in test example 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The following examples are provided to better illustrate the embodiments of the present invention.
Example 1
A microbial composite starter, comprising the following strains:
lactobacillus helveticus L551, lactobacillus paracasei L578, and streptococcus thermophilus S56; wherein the ratio of the number of the viable bacteria of the Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56 is 400:300:35
The Lactobacillus helveticus L551 has the strain preservation number of CGMCC No.15604, the Latin name of Lactobacillus helveticus, the patent CN111685328A, the patent name of Naqu 4580 probiotics and the application of the preparation thereof in food for adjusting emotion, and the strain is preserved in the China general microbiological culture Collection center;
the strain preservation number of the Lactobacillus paracasei L578 is CGMCC No.15603, the Latin name is Lactobacillus paracasei, and the strain disclosed in patent CN111685328A and patent application of Naqu 4580 probiotics and preparations thereof in food for adjusting emotion is preserved in the China general microbiological culture Collection center;
the strain preservation number of the streptococcus thermophilus S56 is CGMCC No.22066, and the Latin name is as follows: streptococcus thermophilus is preserved in China general microbiological culture Collection center at 25.03.2021, and the preservation addresses are as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
The preparation method of the microbial combined leaven comprises the following steps:
a. activating lactobacillus helveticus L551, lactobacillus paracasei L578 and streptococcus thermophilus S56, and then respectively adding the activated lactobacillus helveticus L551, lactobacillus paracasei L578 and streptococcus thermophilus S56 into a liquid culture medium to ferment for 10 hours at 42 ℃ to respectively obtain lactobacillus helveticus fermentation liquor, lactobacillus paracasei fermentation liquor and streptococcus thermophilus fermentation liquor;
b. respectively centrifuging the lactobacillus helveticus fermentation liquor, the lactobacillus paracasei fermentation liquor and the streptococcus thermophilus fermentation liquor, and separating thalli to obtain lactobacillus helveticus bacterial sludge, lactobacillus paracasei bacterial sludge and streptococcus thermophilus bacterial sludge;
c. respectively adding freeze-drying protective agents into the lactobacillus helveticus bacterial sludge, the lactobacillus paracasei bacterial sludge and the streptococcus thermophilus bacterial sludge, and then carrying out freeze drying to obtain lactobacillus helveticus bacterial powder, lactobacillus paracasei bacterial powder and streptococcus thermophilus bacterial powder;
d. compounding the Lactobacillus helveticus bacterium powder, the Lactobacillus paracasei bacterium powder and the Streptococcus thermophilus bacterium powder according to the ratio of the viable count of the Lactobacillus helveticus, the Lactobacillus paracasei and the Streptococcus thermophilus in the microbial composite starter to obtain the microbial composite starter.
Wherein the liquid culture medium comprises the following components in percentage by mass: glucose: 3%, yeast extract powder: 1.5%, soy peptone: 1%, disodium hydrogen phosphate: 0.15%, sodium dihydrogen phosphate: 0.25%, magnesium sulfate: 0.02%, ascorbic acid: 0.3%, tween 80: 0.07 percent.
The freeze-drying protective agent comprises the following components in percentage by mass: 10% of skim milk, 8% of trehalose, 3% of monosodium glutamate, 2% of sucrose, 1% of glycerol, 1% of sodium ascorbate and the balance of sterile water; the addition amount of the freeze-drying protective agent is 100 percent of the mass of the bacterial sludge.
The preparation method of the yoghourt by using the prepared microbial combined starter comprises the following steps:
s1, preparing 80 g of white granulated sugar, 900 g of fresh milk, 5g of concentrated whey powder, 1 g of sodium glutamate and 0.04 g of microbial combined leaven (wherein, the viable count of the Lactobacillus helveticus is 4 multiplied by 10)10cfu/g, viable count of Lactobacillus paracasei 3X 1010cfu/g, Streptococcus thermophilus 3.5X 109cfu/g) and balance water; heating fresh milk to 52 deg.C, adding white sugar, whey powder and sodium glutamate, stirring for dissolving, and heating to 58 deg.C to obtain milk solution;
s2, homogenizing the milk at 60 ℃ and 10Mpa, pasteurizing at 95 ℃ for 330S, cooling to 42 ℃ after pasteurization, adding a microbial combined starter, fermenting at 42 ℃ for 11h, adjusting the pH of the fermentation liquor to 3.8 and the acidity to 115 DEG T, stirring and demulsifying in a fermentation tank, turning over the tank, cooling and refining, filling the obtained yoghourt at 20 +/-1 ℃, sealing, refrigerating at 4 ℃ and aging.
Example 2
A microbial composite starter, comprising the following strains:
lactobacillus helveticus L551, lactobacillus paracasei L578, and streptococcus thermophilus S56; wherein the ratio of the viable count of the Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56 is 350:400: 50.
The preparation method of the microbial combined leaven comprises the following steps:
a. activating lactobacillus helveticus L551, lactobacillus paracasei L578 and streptococcus thermophilus S56, and then respectively adding the activated lactobacillus helveticus L551, lactobacillus paracasei L578 and streptococcus thermophilus S56 into a liquid culture medium to ferment for 12 hours at 37 ℃ to respectively obtain lactobacillus helveticus fermentation liquor, lactobacillus paracasei fermentation liquor and streptococcus thermophilus fermentation liquor;
b. respectively centrifuging the lactobacillus helveticus fermentation liquor, the lactobacillus paracasei fermentation liquor and the streptococcus thermophilus fermentation liquor, and separating thalli to obtain lactobacillus helveticus bacterial sludge, lactobacillus paracasei bacterial sludge and streptococcus thermophilus bacterial sludge;
c. respectively adding freeze-drying protective agents into the lactobacillus helveticus bacterial sludge, the lactobacillus paracasei bacterial sludge and the streptococcus thermophilus bacterial sludge, and then carrying out freeze drying to obtain lactobacillus helveticus bacterial powder, lactobacillus paracasei bacterial powder and streptococcus thermophilus bacterial powder;
d. compounding the Lactobacillus helveticus bacterium powder, the Lactobacillus paracasei bacterium powder and the Streptococcus thermophilus bacterium powder according to the ratio of the viable count of the Lactobacillus helveticus, the Lactobacillus paracasei and the Streptococcus thermophilus in the microbial composite starter to obtain the microbial composite starter.
Wherein the liquid culture medium comprises the following components in percentage by mass: glucose: 2.5%, yeast extract powder: 1%, soy peptone: 0.8%, disodium hydrogen phosphate: 0.13%, sodium dihydrogen phosphate: 0.2%, magnesium sulfate: 0.01%, ascorbic acid: 0.2%, tween 80: 0.06% and the balance sterile water.
The freeze-drying protective agent comprises the following components in percentage by mass: 8% of skim milk, 5% of trehalose, 0.5% of monosodium glutamate, 1% of sucrose, 0.5% of glycerol, 0.1% of sodium ascorbate and the balance of sterile water; the addition amount of the freeze-drying protective agent is 85 percent of the mass of the bacterial sludge.
The preparation method of the yoghourt by using the prepared microbial combined starter comprises the following steps:
s1, preparing 110 g of white granulated sugar, 850 g of fresh milk, 2g of concentrated whey powder, 0.5g of sodium glutamate and 0.01 g of microbial combined leaven (wherein, the viable count of the Lactobacillus helveticus is 3.5 multiplied by 10)10cfu/g, viable count of Lactobacillus paracasei 4X 1010cfu/g, Streptococcus thermophilus 5X 109cfu/g) and balance water; heating fresh milk to 50 deg.C, adding white sugar, whey powder and sodium glutamate, stirring for dissolving, and heating to 55 deg.C to obtain milk solution;
s2, homogenizing the milk at 50 ℃ and 20Mpa, pasteurizing at 94 ℃ for 300S, cooling to 37 ℃ after pasteurization, adding a microbial combined starter, fermenting at 37 ℃ for 15h, adjusting the pH of the fermentation liquor to 3.9 and the acidity to 114 DEG T, stirring and demulsifying in a fermentation tank, turning over the tank, cooling and refining, filling the obtained yoghourt at 20 +/-1 ℃, sealing, refrigerating at 4 ℃ and aging.
Example 3
A microbial composite starter, comprising the following strains:
lactobacillus helveticus L551, lactobacillus paracasei L578, and streptococcus thermophilus S56; wherein the ratio of the viable count of the Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56 is 450:200: 20.
The preparation method of the microbial combined leaven comprises the following steps:
a. activating lactobacillus helveticus L551, lactobacillus paracasei L578 and streptococcus thermophilus S56, and then respectively adding the activated lactobacillus helveticus L551, lactobacillus paracasei L578 and streptococcus thermophilus S56 into a liquid culture medium to ferment for 10 hours at 45 ℃ to respectively obtain lactobacillus helveticus fermentation liquor, lactobacillus paracasei fermentation liquor and streptococcus thermophilus fermentation liquor;
b. respectively centrifuging the lactobacillus helveticus fermentation liquor, the lactobacillus paracasei fermentation liquor and the streptococcus thermophilus fermentation liquor, and separating thalli to obtain lactobacillus helveticus bacterial sludge, lactobacillus paracasei bacterial sludge and streptococcus thermophilus bacterial sludge;
c. respectively adding freeze-drying protective agents into the lactobacillus helveticus bacterial sludge, the lactobacillus paracasei bacterial sludge and the streptococcus thermophilus bacterial sludge, and then carrying out freeze drying to obtain lactobacillus helveticus bacterial powder, lactobacillus paracasei bacterial powder and streptococcus thermophilus bacterial powder;
d. compounding the Lactobacillus helveticus bacterium powder, the Lactobacillus paracasei bacterium powder and the Streptococcus thermophilus bacterium powder according to the ratio of the viable count of the Lactobacillus helveticus, the Lactobacillus paracasei and the Streptococcus thermophilus in the microbial composite starter to obtain the microbial composite starter.
Wherein the liquid culture medium comprises the following components in percentage by mass: glucose: 3.5%, yeast extract powder: 2%, soy peptone: 1.2%, disodium hydrogen phosphate: 0.17%, sodium dihydrogen phosphate: 0.3%, magnesium sulfate: 0.03%, ascorbic acid: 0.4%, tween 80: 0.08% and the balance sterile water.
The freeze-drying protective agent comprises the following components in percentage by mass: 15% of skim milk, 12% of trehalose, 5% of monosodium glutamate, 5% of sucrose, 3% of glycerol, 3% of sodium ascorbate and the balance of sterile water; the addition amount of the freeze-drying protective agent is 200 percent of the mass of the bacterial sludge.
The preparation method of the yoghourt by using the prepared microbial combined starter comprises the following steps:
s1, preparing 50 g of white granulated sugar, 920 g of fresh milk, 10 g of concentrated whey powder, 2g of sodium glutamate and 0.08 g of microbial combined leaven for each liter of yoghourt product (wherein the viable count of lactobacillus helveticus is 4.5 multiplied by 10)10cfu/g, viable count of Lactobacillus paracasei 2X 1010cfu/g, Streptococcus thermophilus 2X 109cfu/g) and balance water; heating fresh milk to 55 deg.C, adding white sugar, whey powder and sodium glutamate, stirring for dissolving, and heating to 60 deg.C to obtain milk solution；
S2, homogenizing the milk at 75 ℃ and 5Mpa, pasteurizing at 96 ℃ for 350S, cooling to 45 ℃ after pasteurization, adding a microbial combined starter, fermenting at 45 ℃ for 5h, adjusting the pH of the fermentation liquor to 3.9 and the acidity to 110 DEG T, stirring and demulsifying in a fermentation tank, turning over the tank, cooling and refining, filling the obtained yoghourt at 20 +/-1 ℃, sealing, refrigerating at 4 ℃ and aging.
Test example 1
1. The aroma of the yogurt obtained in example 1 was detected and analyzed, and the analysis method and analysis result were as follows:
GC-MS detection method for detecting aroma components
The instrument comprises the following steps: 7890A gas chromatograph-5975C mass spectrometer (Agilent, USA); solid Phase Microextraction (SPME) hand sample handle, 50/30 μm DVB/CAR/PDMS extraction head, 0.75mm I.D.SPME special liner (Supelco Corp.); 5078HW-1 digital display constant temperature magnetic stirrer (Hangzhou instrument motor Co., Ltd.); model JJ200 electronic balance (jimmy test instruments).
Chromatographic conditions are as follows: a chromatographic column: HP-5 polymethylsiloxane quartz capillary column; flow rate of He gas: 1.0 ml/min; temperature rising procedure: the initial temperature is 35 ℃, the temperature is kept for 6min, then the temperature is increased to 90 ℃ at the speed of 3 ℃/min, then the temperature is increased to 150 ℃ at the speed of 5 ℃/min, finally the temperature is increased to 210 ℃ at the speed of 10 ℃/min, and the temperature is kept for 10 min; sample inlet temperature: 250 deg.C (applicable to DVB/CAR/PDMS extraction head).
Mass spectrum conditions: EI ion source: electron bombardment energy 70 eV; ion source temperature: 230 ℃ to 230 ℃.
headspace-SPME method: taking 5ml of yogurt sample, adding 1ml of saturated NaCl solution, extracting at 60 ℃ for 30min by using 50/30 mu m DVB/CAR/PDMS extraction head headspace at constant temperature, then thermally analyzing the extraction head at 250 ℃ for 5min, then injecting and carrying out GC-MS analysis.
The detection result is shown in fig. 1, the fragrance of the yogurt obtained in example 1 is obviously different from that of a common starter, the yogurt takes butanedione and acetoin as main fragrant substances, the content of volatile acids is the highest, and acetic acid is taken as a representative, and a certain amount of caproic acid, caprylic acid and capric acid are used. The butanedione is cream fragrance, and the 2-heptanone is blue and sweet cheese fragrance. Therefore, the yoghourt has strong cheese fragrance with acetic acid flavor.
2. Detection of acetic acid content
Acetic acid is a short-chain fatty acid, is a metabolite of probiotics in intestinal tracts, can reduce the pH value in the intestinal tracts, promote the absorption of calcium and magnesium ions in the intestinal tracts, inhibit the infection of harmful bacteria and stimulate the intestinal tract to creep so as to relieve constipation; it also has effects in improving intestinal permeability and barrier function of intestinal mucosa, and enhancing intestinal immunity.
2.1 test materials: the yogurt obtained in example 1, the glacial acetic acid standard, the commercially available yogurt 1 fermented from the commercially available yogurt starter (commercially available yogurt starter one: Streptococcus thermophilus, Lactobacillus bulgaricus, Lactobacillus paracasei), and the commercially available yogurt 2 fermented from the commercially available yogurt starter two (commercially available yogurt starter two: Streptococcus thermophilus, Lactobacillus bulgaricus, Lactobacillus paracasei, Bifidobacterium lactis, Lactobacillus acidophilus).
2.2 test equipment: agilent Technologies 1260Infinity II
2.3 sample pretreatment: diluting the sample by 4 times, centrifuging at 9000 r/min for 10min, filtering with a membrane, and injecting.
2.4 test results:
2.4.1 preparation of Standard Curve
The acetic acid content of the standards is shown in table 1.
TABLE 1
The resulting standard curve is shown in FIG. 2. The standard product spectrum peaks at about 4.7min, as shown in FIG. 3.
2.4.2 the spectrum of the yogurt obtained in example 1 was determined as shown in fig. 4, and the peak area in the spectrum was substituted into the standard curve formula of the acetic acid content (y: 900667x +25.246), and the acetic acid content was calculated to be 0.38 g/L.
However, no target peak appeared around 4.7min in the current commercial yoghurt (as shown in fig. 5 and 6), i.e. no acetic acid could be detected in the commercial starter fermented milk under the test conditions.
2.5 conclusion of the test
The fermented yoghurt obtained in example 1 was subjected to detection of acetic acid by liquid chromatography, and the acetic acid content was 0.38 g/L. And the detection of acetic acid is carried out on the two types of the commercial starter fermented yoghurts by the same method, and the detection result is as follows: acetic acid was not detected in both fermented milks.
3. Gamma aminobutyric acid content detection
3.1 advantages of Strain combination
The preparation of the yoghurt was carried out by setting up the starter cultures composed of different strains, the preparation method was the same as in example 1, and the contents of γ -aminobutyric acid in the yoghurts obtained with different starter cultures are shown in table 2.
TABLE 2
Note: the total number of viable bacteria of each group of the leavening agents is basically equivalent to that of the leavening agents in the embodiment 1, and the ratio of the number of viable bacteria of the strains in the leavening agents related to the strain combination is the same as that of the embodiment 1.
As can be seen from table 2, the amount of γ -aminobutyric acid produced in the yogurt produced by the combination of the three strains in the microbial composite starter in example 1 is the largest, and the yogurt is stable in taste and flavor by the combination.
3.2 liquid-phase detection and analysis of the content of gamma-aminobutyric acid in the yogurt
The chromatogram obtained by performing liquid phase detection analysis on the yogurt in example 1 is shown in fig. 7, and the calculated content of gamma-aminobutyric acid in the yogurt is 800 ug/g.
3.3 relationship between increase in acidity and content of gamma-aminobutyric acid
In the fermentation process of the yogurt in example 1, 1.0g/L of sodium glutamate was added. And (3) determining the change condition of the gamma-aminobutyric acid under different acidity. The content of the gamma-aminobutyric acid is increased along with the increase of the fermentation acidity, and when the acidity of the yoghourt reaches 150 DEG T, the content of the gamma-aminobutyric acid reaches the maximum. The increase of the gamma-aminobutyric acid tends to be smooth when the fermentation time is about 11 hours, and meanwhile, the sour-sweet ratio of the yoghourt is relatively suitable for consumers to eat, so that the yoghourt is fermented to about 115 DEG T when the fermentation time is 11 hours, and the yoghourt is suitable for the acidity of high-yield gamma-aminobutyric acid.
3.4 influence of different sodium glutamate additions on the content of gamma-aminobutyric acid
Performing a fresh milk fermentation test by using the microbial combined starter in the embodiment 1, and adding 0.125g/L, 0.25g/L, 0.5g/L, 1.0g/L, 1.5g/L, 2.0g/L, 3.0g/L and 5.0g/L of sodium glutamate respectively, wherein the fermentation method is the same as that of the embodiment 1, the fermentation is terminated when the temperature reaches 110 ℃ T, and the detection of gamma-aminobutyric acid is performed, and the result shows that different contents of sodium L-glutamate have certain influence on the content of gamma-aminobutyric acid, and when the content of sodium L-glutamate is lower, the content of gamma-aminobutyric acid increases along with the increase of the content of sodium L-glutamate; when the content of the sodium L-glutamate reaches a certain value (1.5g/L), the content of the gamma-aminobutyric acid is slightly changed, which is probably because other factors in the fermentation liquor limit the conversion of the lactic acid bacteria on the sodium L-glutamate. Therefore, 1.5g/L was selected as the optimum sodium L-glutamate addition concentration.
3.5 the aroma components of the yoghurts prepared in examples 2 and 3 were measured, and the main aroma components were the same as in example 1.
Meanwhile, the contents of acetic acid and gamma-aminobutyric acid in the yoghurts prepared in examples 2 and 3 were measured. Through detection, the content of acetic acid in the yoghourt prepared in the example 2 is 0.32g/L, and the content of gamma-aminobutyric acid is 500 ug/g. The content of acetic acid in the yogurt prepared in example 3 was 0.35g/L, and the content of gamma-aminobutyric acid was 700 ug/g.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A microbial combined starter which is characterized in that: the following strains are included:
lactobacillus helveticus L551, lactobacillus paracasei L578, and streptococcus thermophilus S56;
the strain preservation number of the Lactobacillus helveticus L551 is CGMCC No. 15604;
the strain preservation number of the lactobacillus paracasei L578 is CGMCC No. 15603;
the strain preservation number of the streptococcus thermophilus S56 is CGMCC No. 22066.
2. The microbial starter culture of claim 1, wherein: in the microbial combined starter, the ratio of the viable count of the Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56 is 350-450:200-400: 20-50.
3. The method for producing a microbial composite fermentation product according to claim 1 or 2, wherein: the method comprises the following steps:
a. activating the Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56, and then respectively adding the activated Lactobacillus helveticus L551, the Lactobacillus paracasei L578 and the Streptococcus thermophilus S56 into a liquid culture medium for fermentation at 37-45 ℃ for 10-12h to obtain Lactobacillus helveticus fermentation liquid, Lactobacillus paracasei fermentation liquid and Streptococcus thermophilus fermentation liquid;
b. respectively centrifuging the lactobacillus helveticus fermentation liquor, the lactobacillus paracasei fermentation liquor and the streptococcus thermophilus fermentation liquor, and separating thalli to obtain lactobacillus helveticus bacterial mud, lactobacillus paracasei bacterial mud and streptococcus thermophilus bacterial mud;
c. respectively adding freeze-drying protective agents into the lactobacillus helveticus bacterial mud, the lactobacillus paracasei bacterial mud and the streptococcus thermophilus bacterial mud, and then carrying out freeze drying to obtain lactobacillus helveticus bacterial powder, lactobacillus paracasei bacterial powder and streptococcus thermophilus bacterial powder;
d. and compounding the lactobacillus helveticus bacterium powder, the lactobacillus paracasei bacterium powder and the streptococcus thermophilus bacterium powder according to the proportion of the viable count of the lactobacillus helveticus, the lactobacillus paracasei and the streptococcus thermophilus in the microbial combined starter to obtain the microbial combined starter.
4. The method for preparing a microbial starter culture according to claim 3, wherein: in the step a, the liquid culture medium comprises the following components in percentage by mass: glucose: 2.5-3.5%, yeast extract powder: 1-2%, soy peptone: 0.8-1.2%, disodium hydrogen phosphate: 0.13-0.17%, sodium dihydrogen phosphate: 0.2-0.3%, magnesium sulfate: 0.01-0.03%, ascorbic acid: 0.2-0.4%, tween 80: 0.06-0.08% and the balance of sterile water.
5. The method for preparing a microbial starter culture according to claim 3, wherein: the freeze-drying protective agent comprises the following components in percentage by mass: 8-15% of skim milk, 5-12% of trehalose, 0.5-5% of monosodium glutamate, 1-5% of sucrose, 0.5-3% of glycerol, 0.1-3% of sodium ascorbate and the balance of sterile water; the addition amount of the freeze-drying protective agent is 85-200% of the mass of the bacterial sludge.
6. Use of the microbial starter combination according to claim 1 or 2 as a starter for ready-to-use yoghurt.
7. A preparation method of yoghourt is characterized by comprising the following steps: the method comprises the following steps:
s1, heating fresh milk to 50-55 ℃, adding white granulated sugar, whey powder and sodium glutamate, stirring for dissolving, and heating to 55-60 ℃ to obtain milk liquid;
s2, homogenizing and pasteurizing the milk, cooling to 37-45 ℃, adding the microbial combined starter according to claim 1 or 2, fermenting at 37-45 ℃, and stirring for demulsification, cooling and refining when pH of the fermented liquid is less than or equal to 4 and acidity is more than or equal to 110 ℃ T to obtain the yoghourt.
8. A process for preparing yoghurt as claimed in claim 7, characterized in that: in the step S1, the mass ratio of the fresh milk, the white granulated sugar, the whey powder and the sodium glutamate is 850-110: 2-10: 0.5-2; and/or
In step S2, the mass ratio of the microbial composite starter to the fresh milk is 0.01-0.08: 850-920.
9. A process for preparing yoghurt as claimed in claim 7, characterized in that: in step S2, the homogenizing temperature is 50-75 deg.C and the pressure is 5-20 Mpa; and/or
In step S2, the temperature of pasteurization is 94-96 deg.C and the time is 300-350S.
10. A yogurt, characterized by: the method for preparing the yoghourt according to any one of claims 7-9.
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