CN105532876B - Buffalo milk lactic acid milk rich in gamma-aminobutyric acid and preparation method thereof - Google Patents
Buffalo milk lactic acid milk rich in gamma-aminobutyric acid and preparation method thereof Download PDFInfo
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- 235000020246 buffalo milk Nutrition 0.000 title claims abstract description 149
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 title claims abstract description 94
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 title claims abstract description 47
- 229960003692 gamma aminobutyric acid Drugs 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
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- 239000008267 milk Substances 0.000 title claims description 12
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title description 14
- 235000014655 lactic acid Nutrition 0.000 title description 7
- 239000004310 lactic acid Substances 0.000 title description 7
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- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 48
- 235000014897 Streptococcus lactis Nutrition 0.000 claims abstract description 42
- 241000194020 Streptococcus thermophilus Species 0.000 claims abstract description 37
- 241000894006 Bacteria Species 0.000 claims abstract description 35
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 2
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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
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1322—Inorganic compounds; Minerals, including organic salts thereof, oligo-elements; Amino-acids, peptides, protein-hydrolysates or derivatives; Nucleic acids or derivatives; Yeast extract or autolysate; Vitamins; Antibiotics; Bacteriocins
-
- 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
- A23V2400/169—Plantarum
-
- 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
- A23V2400/231—Lactis
-
- 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 buffalo milk yoghourt rich in gamma-aminobutyric acid (GABA) and a preparation method thereof, belongs to the technical field of dairy product processing, and is prepared by mixing three bacteria, namely lactococcus lactis subspecies lactis capable of producing GABA, lactobacillus plantarum and traditional yoghourt strain streptococcus thermophilus, which are separated from buffalo milk. The preparation process is simple, and the prepared yoghourt is rich in frankincense, sticky in texture, fine and smooth in taste and rich in nutrition; the GABA content is high and can reach 3.6g/kg at most, and the GABA has various physiological functions of reducing blood pressure, resisting anxiety, resisting depression, improving sleep, detoxifying and the like; and no additive is added, so that the quality guarantee period is long, and the method is safe, environment-friendly and healthy.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to the technical field of dairy product processing, in particular to buffalo milk yoghurt rich in gamma-aminobutyric acid and a preparation method thereof.
[ technical background ] A method for producing a semiconductor device
Buffalo milk is a special dairy product in southern China, and has the characteristics of high quality, safety and few properties. The buffalo milk has low yield, but the protein, the milk fat, the vitamins, the trace elements and the like contained in the buffalo milk are higher than those contained in the black and white milk, and are known as the king of milk. At present, the buffalo milk processing industry in China is in a primary stage, mainly liquid milk is used as main material, and high value-added products are few.
The yoghourt is a product prepared by taking cow milk as a raw material, adding lactobacillus after pasteurization and cooling, and fermenting, wherein the fermented milk contains a large amount of probiotics and has the effects of regulating gastrointestinal flora, improving the immune system of the organism and the like, so the yoghourt is popular and has great market development potential.
Gamma-aminobutyric acid (GABA for short) is a natural non-protein amino acid, and a large number of researches show that GABA has various physiological functions of reducing blood pressure, resisting convulsion, easing pain, improving sleep, detoxifying and the like. The buffalo milk lactic acid and GABA can be a perfect combination, and the buffalo milk yoghourt rich in GABA is obtained by utilizing the fermentation of the lactic acid bacteria, so that the nutritive value of the yoghourt can be further improved, and the buffalo milk yoghourt has more physiological effects in GABA so as to meet the high requirements of people on functional foods, be applied to market development and be the problem which is urgently needed to be solved by the buffalo milk processing industry at present.
[ summary of the invention ]
In view of the problems provided above, the invention provides buffalo milk rich in gamma-aminobutyric acid and a preparation method thereof, in order to make up the defects of the existing buffalo milk functional product and production technology.
The inventor discovers that lactococcus lactis subsp.lactis 25# separated from fresh buffalo milk is preserved in China center for type culture collection, CCTCC for short, and is stored in China center for type culture collection, namely Wuhan university, Wuhan city, Hubei province, China, with the preservation date of 2015.11.25 and the preservation number of CCTCC NO of M2015700, and Lactobacillus plantarum 12# and is preserved in China center for type culture collection, namely CCTCC for short, and is stored in Wuhan university, Wuhan city, Hubei province, China, with the preservation date of 2015.11.25 and the preservation number of CCTCC NO of M2015699, both strains can produce gamma-aminobutyric acid and have higher yield, so the two strains can be applied to fermentation of buffalo milk to prepare the buffalo milk rich in gamma-aminobutyric acid. The technical scheme of the invention is as follows:
the sour milk rich in gamma-aminobutanes is prepared by fermenting skim buffalo milk serving as a main material with 7-9% of white granulated sugar, 0.4-0.9% of sodium glutamate and 5-10% of composite strains in the total mass of the skim buffalo milk, wherein the preferable adding amount of the white granulated sugar, the sodium glutamate and the composite strains are respectively 8%, 0.6% and 7% of the total mass of the skim buffalo milk; the composite strain is formed by mixing activated lactococcus lactis subsp lactis, lactobacillus plantarum and streptococcus thermophilus, wherein the lactococcus lactis subsp lactis and the lactobacillus plantarum are obtained by separating from raw buffalo milk and can be called as 'derived from buffalo milk and used for buffalo milk'; the lactococcus lactis subsp. lactis obtained by separation is lactococcus lactis subsp. 25#, is preserved in China center for type culture collection in 11 months and 25 days 2015, and has a preservation number of CCTCCNO: M2015700; the Lactobacillus plantarum obtained by separation is Lactobacillus plantarum (Lactobacillus plantarum) No. 12, which is preserved in China center for type culture Collection in 2015, 11 and 25 months, with the preservation number of CCTCC NO: M2015699, while streptococcus thermophilus (with the preservation number of CGMCC NO:2718) is purchased from China center for general microbiological culture collection.
The skim buffalo milk is prepared by filtering and purifying qualified fresh buffalo milk and then carrying out cream separation, wherein the mass percent of protein in the skim buffalo milk is 5.5-6.8%, the mass percent of fat is 0.2-0.5%, and the acidity is less than or equal to 16 DEG T. In addition, preferably, the composite bacterial strain is prepared by activated lactococcus lactis subsp lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 1-2: 1: 1-2, mixing; further preferably, the composite strain is prepared by activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 2: 1: 1 are mixed.
The composite strain is prepared by respectively activating lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus and then mixing the activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to a mass ratio, and the specific activating preparation method comprises the following steps: respectively performing continuous secondary recovery subculture on lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus in an MRS slant culture medium to obtain strains; then respectively inoculating the strains into 100ml of MRS broth culture medium, and performing shake culture at 35-40 ℃ for 20-24h to obtain culture bacteria liquid; centrifuging the culture solution for 6-10min at 4800-; and finally, uniformly mixing the three target bacteria liquids according to the proportion, inoculating the three target bacteria liquids into the sterilized skim buffalo milk according to the amount of 20 percent of the total mass of the skim buffalo milk, and placing the skim buffalo milk in a shaking table at 37 ℃ for culturing for 16-24 hours to obtain the composite strain.
The buffalo milk yogurt provided by the invention is set yogurt. Wherein the viable count of lactobacillus is not less than 2.2 × 108CFU·mL-1The protein is more than or equal to 4.9 percent, the fat is less than or equal to 0.5 percent, the acidity is more than or equal to 70 degrees T, the GABA content can reach 3.6g/kg at most, the sanitary indexes all meet the requirements of GB19302-2010 fermented milk, and the product has good sense.
The preparation method of the buffalo milk lactic acid rich in gamma-aminobutyric acid comprises the following steps:
1) heating skim buffalo milk to 60-80 ℃, adding white granulated sugar and sodium glutamate according to the proportion, uniformly stirring, heating to 80-95 ℃, sterilizing for 10-12min, and cooling to 35-37 ℃;
2) inoculating the composite strain into the skim buffalo milk treated in the step 1), fully stirring uniformly and then bottling;
3) fermenting the skimmed buffalo milk filled in the step 2) at the constant temperature of 38-41 ℃ for 3-5h until curd is formed, and taking the pH value of the skimmed buffalo milk as the fermentation end point when the pH value is 4.0-4.5 to obtain fermented yoghourt;
4) putting the fermented yoghourt obtained in the step 3) into a refrigerated cabinet with the temperature of 4-6 ℃, and after-ripening for 22-24h to obtain the finished product of the buffalo milk.
The invention provides buffalo milk yoghourt rich in gamma-aminobutyric acid and a preparation method thereof, and compared with the prior art, the buffalo milk yoghourt has the following beneficial effects:
1. the invention mixes two strains of lactococcus lactis subsp lactis and lactobacillus plantarum which are separated from buffalo milk and produce gamma-aminobutyric acid with a traditional yogurt strain streptococcus thermophilus to ferment the buffalo milk synergistically, and simultaneously adds a certain amount of sodium glutamate to promote the lactococcus lactis subsp lactis and the lactobacillus plantarum to enrich and produce more GABA, and prepares the buffalo milk rich in gamma-aminobutyric acid by changing process conditions and fermentation strains, wherein the GABA content in the yogurt can reach 3.6g/kg at most, so that the buffalo milk yogurt not only has the nutritional value of common yogurt, but also increases various physiological functions of blood pressure reduction, convulsion resistance, pain relief, detoxification and the like of GABA, and further improves the nutritional and health-care effects of the buffalo milk yogurt as a whole.
2. The invention relates to a method for separating lactococcus lactis subspecies lactis and lactobacillus plantarum from buffalo milk, wherein the lactococcus lactis subspecies lactis and lactobacillus plantarum are separated from the buffalo milk, and the commercially available streptococcus thermophilus are separated from the buffalo milk according to the mass ratio of 1-2: 1: 1-2, preparing a composite strain for buffalo milk fermented yoghourt, and overcoming the defects of over-viscous curd and over-hard tissue state of the buffalo milk in the fermentation process; moreover, the inventor gropes a plurality of experiments, the fermentation temperature is kept constant at 37-41 ℃, and is 3-4 ℃ lower than the fermentation temperature (40-45 ℃) adopted by the conventional fermented milk, so that the prepared yoghurt has strong frankincense, sticky texture and fine and smooth taste; in addition, because the lactobacillus plantarum has the capabilities of strongly producing acid and maintaining the balance of flora, the number of live lactobacillus in the yoghourt is higher, and is 5-8 times higher than that of the common yoghourt through detection, the acidification degree of the yoghourt is controllable, and the quality guarantee period of the product can be effectively prolonged.
3. The invention provides a preparation method of buffalo milk lactate rich in gamma-aminobutyric acid, which has simple process, is environment-friendly and has no additive, and the advantages of the buffalo milk lactate rich in gamma-aminobutyric acid are mainly shown as follows: (1) the invention adopts three bacteria of lactococcus lactis subspecies lactis, lactobacillus plantarum and traditional yogurt strain streptococcus thermophilus which are separated from buffalo milk to mix and ferment the yogurt, so that the obtained yogurt has the characteristics of viscosity, rich and safe frankincense and zero additive, and the application of a large amount of chemical raw materials such as thickening agent, essence and the like in the buffalo milk yogurt is reduced. (2) The strain is in a direct vat set mode, and the mixed strain is only required to be added into the sterilized skim buffalo milk to ferment the yoghourt at constant temperature, so that the process is simple, the requirement on production conditions is not high, and industrialization can be realized.
[ detailed description ] embodiments
The following examples may assist those skilled in the art in a more complete understanding of the present invention, but are not intended to limit the invention in any way.
In the following examples, the proportions are by weight unless otherwise specified.
In the following examples, the detection methods involved are as follows:
the fat determination method comprises the following steps: reference is made to GB 5413.3;
the protein determination method comprises the following steps: reference is made to GB 5009.5;
the acidity determination method comprises the following steps: reference is made to GB 5413.34-2010;
the viable count method of the lactobacillus comprises the following steps: reference GB 2746-;
method for determining gamma-aminobutyric acid (GABA): high Performance Liquid Chromatography (HPLC).
The following examples are examples of sources of raw materials and equipment:
in the following examples, the compound strain used was prepared by mixing activated lactococcus lactis subsp lactis, Lactobacillus plantarum and streptococcus thermophilus, wherein lactococcus lactis subsp lactis 25# isolated from fresh buffalo milk is deposited in China Center for Type Culture Collection (CCTCC) with the address of Wuhan university, Wuhan, Hubei province, China, with the preservation number of M2015700 and the preservation date of 2015.11.25; the lactobacillus plantarum is lactobacillus plantarum 12# separated from fresh buffalo milk, is preserved in China center for type culture Collection (CCTCC for short), is named as Wuhan university in Wuhan City, Hubei province, China, has a preservation number of M2015699 and a preservation date of 2015.11.25; the streptococcus thermophilus is CGMCC NO:1.2718, and is purchased from China general microbiological culture Collection center.
Buffalo milk, a buffalo farm of the research institute of buffalo in Guangxi; MRS medium, bio-technology ltd, garden haibo, celadon; MRS broth, guangdong cika microbiology ltd; white granulated sugar, south Ning molasses commerce and trade company; low-speed large capacity centrifuge: the model is as follows: DD-5M; a freeze dryer: ALPHA 1-2LDplus type, Germany MARTIN CHRIST; the three-purpose constant temperature water tank model: DK-600S, Shanghai sperm macro laboratory Equipment Co., Ltd; electronic balance ME204E/02 Mettler-Torledo instruments (Shanghai) Inc.; cup sealing machine, model: ET-D6, E Dong electromechanical Co., Ltd, Guangzhou; fermentation tank: the model is as follows: fermentar 300 series 2L, Inc. of UK (HT Mini-Fermentar/Bioreacor).
Example 1
The buffalo milk yoghourt rich in gamma-aminobutyric acid is set yoghourt. The yoghourt is prepared by taking skim buffalo milk as a main material and adding 7% of white granulated sugar, 0.4% of sodium glutamate and 5% of composite strain by the total mass of the skim buffalo milk for fermentation. The skim buffalo milk is prepared by filtering and purifying qualified fresh buffalo milk and then performing cream separation, wherein the protein content is 5.5%, the fat content is 0.5%, and the acidity is less than or equal to 16 DEG T. The composite strain is prepared by activating lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 1: 1: 1, the specific activating preparation method comprises the following steps: respectively carrying out continuous secondary revival subculture on the freeze-dried lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus in an MRS slant culture medium to obtain strains; then respectively inoculating the strains into 100ml of MRS broth culture medium, and carrying out shake cultivation for 24h at 35 ℃ to obtain culture bacteria liquid; centrifuging the cultured bacteria liquid with a centrifuge at 4800rpm for 10min, and collecting supernatant to obtain target bacteria liquid, i.e. activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus; and finally, uniformly mixing the three target bacteria liquids according to the proportion, inoculating the three target bacteria liquids into the sterilized skim buffalo milk, wherein the inoculation amount is 20% of the total mass of the skim buffalo milk, and then placing the skim buffalo milk in a shaking table at 37 ℃ for culturing for 16h to obtain the composite strain.
The preparation method of the yoghourt comprises the following steps: heating skim buffalo milk to 60 ℃, then adding 7% of white granulated sugar and 0.4% of sodium glutamate, uniformly stirring, heating to 85-90 ℃, sterilizing for 12min, and cooling to 35 ℃ for later use; inoculating the composite strain into the skim buffalo milk treated by the method, wherein the inoculation amount is 5% of the total mass of the skim buffalo milk, fully stirring uniformly and then bottling; then placing the skimmed buffalo milk filled in the cup at the constant temperature of 38 ℃ for fermenting for 3-5h until the milk curd is formed, and regarding the fermentation end point when the pH value is 4.0-4.5 to obtain fermented yoghourt; and finally, putting the fermented yoghourt into a refrigerated cabinet with the temperature of 4 ℃, and after-ripening for 24 hours to obtain the finished product of the buffalo milk.
Example 2
The buffalo milk yoghourt rich in gamma-aminobutyric acid is set yoghourt. The yoghourt takes skim buffalo milk as a main material, and is prepared by adding 8% of white granulated sugar, 0.6% of sodium glutamate and 7% of composite strain in the total mass of the skim buffalo milk and fermenting. The skim buffalo milk is prepared by filtering and purifying qualified fresh buffalo milk and then performing cream separation, wherein the protein content is 5.82%, the fat content is 0.4%, and the acidity is less than or equal to 16 DEG T. The composite strain is prepared by activating lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 2: 1: 1, the specific activation preparation method comprises the following steps: respectively carrying out continuous twice revival subculture on lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus stored in the glycerinum tube in an MRS slant culture medium to obtain strains; then respectively inoculating the strains into 100ml of MRS broth culture medium, and carrying out shake cultivation for 22h at 37 ℃ to obtain culture bacteria liquid; centrifuging the culture bacteria liquid for 8min at 5000rpm by a centrifuge, and collecting supernatant to obtain target bacteria liquid, namely activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus; and finally, uniformly mixing the three target bacteria liquids according to the proportion, inoculating the three target bacteria liquids into the sterilized skim buffalo milk, wherein the inoculation amount is 20 percent of the total mass of the skim buffalo milk, and then placing the skim buffalo milk in a shaking table at 37 ℃ for 18 hours to obtain the composite strain.
The preparation method of the yoghourt comprises the following steps: heating skim buffalo milk to 70 ℃, then adding 8% of white granulated sugar and 0.6% of sodium glutamate, uniformly stirring, heating to 90-95 ℃, sterilizing for 10min, and cooling to 37 ℃ for later use; inoculating the composite strain into the treated skim buffalo milk, wherein the inoculation amount is 7 percent of the total mass of the skim buffalo milk, fully stirring uniformly and then bottling; then placing the skimmed buffalo milk filled in the cup at the constant temperature of 41 ℃ for fermenting for 3-5h until the milk curd is formed, and regarding the fermentation end point when the pH value is 4.0-4.5 to obtain fermented yoghourt; and finally, putting the fermented yoghourt into a refrigerated cabinet with the temperature of 6 ℃, and after-ripening for 22 hours to obtain the finished buffalo milk.
Example 3
The buffalo milk yoghourt rich in gamma-aminobutyric acid is set yoghourt. The yoghourt takes skim buffalo milk as a main material, and is prepared by adding 9% of white granulated sugar, 0.9% of sodium glutamate and 10% of composite strain in the total mass of the skim buffalo milk and fermenting. The skim buffalo milk is prepared by filtering and purifying qualified fresh buffalo milk and then carrying out milk fat separation, wherein the protein content is 6.8%, the fat content is 0.5%, and the acidity is less than or equal to 16 DEG T. The composite strain is prepared by activating lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 1: 1: 2, the specific activating preparation method comprises the following steps: respectively carrying out continuous twice revival subculture on freeze-dried lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus in an MRS slant culture medium to obtain strains; then respectively inoculating the strains into 100ml of MRS broth culture medium, and performing shake cultivation for 20h at 40 ℃ to obtain culture bacteria liquid; centrifuging the culture solution with a centrifuge at 5200rpm for 6min, and collecting supernatant to obtain target bacteria solution, i.e. activated lactococcus lactis subspecies lactis, Lactobacillus plantarum and Streptococcus thermophilus; and finally, uniformly mixing the three target bacteria liquids according to the proportion, inoculating the three target bacteria liquids into the sterilized skim buffalo milk, wherein the inoculation amount is 20 percent of the total mass of the skim buffalo milk, and then placing the skim buffalo milk in a shaking table at 37 ℃ for culturing for 20 hours to obtain the composite strain.
The preparation method of the yoghourt comprises the following steps: heating skim buffalo milk to 80 ℃, then adding 9% of white granulated sugar and 0.9% of sodium glutamate, uniformly stirring, heating to 80-90 ℃, sterilizing for 11min, and cooling to 36 ℃ for later use; inoculating the composite strain into the treated skim buffalo milk, wherein the inoculation amount is 10% of the total mass of the skim buffalo milk, fully stirring uniformly and then bottling; then placing the skimmed buffalo milk filled in the cup at the constant temperature of 40 ℃ for fermenting for 3-5h until the milk curd is formed, and regarding the fermentation end point when the pH value is 4.0-4.5 to obtain fermented yoghourt; and finally, putting the fermented yoghourt into a refrigerated cabinet with the temperature of 5 ℃, and after-ripening for 23 hours to obtain the finished product of the buffalo milk.
Example 4
The buffalo milk yoghourt rich in gamma-aminobutyric acid is set yoghourt. The yoghourt takes skim buffalo milk as a main material, and is prepared by adding 9% of white granulated sugar, 0.7% of sodium glutamate and 10% of composite strain in the total mass of the skim buffalo milk and fermenting. The skim buffalo milk is prepared by filtering and purifying qualified fresh buffalo milk and then carrying out milk fat separation, wherein the protein content is 6.65%, the fat content is 0.2%, and the acidity is less than or equal to 16 DEG T. The composite strain is prepared by activating lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 2: 1: 2, the specific activating preparation method comprises the following steps: continuously carrying out secondary revival subculture on lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus stored in the glycerinum tube in an MRS slant culture medium respectively to obtain strains; then respectively inoculating the strains into 100ml of MRS broth culture medium, and performing shake cultivation for 23h at 36 ℃ to obtain culture bacteria liquid; centrifuging the cultured bacteria liquid for 7min at 5100rpm by a centrifugal machine respectively, and collecting supernatant to obtain target bacteria liquid, namely activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus; and finally, uniformly mixing the three target bacteria liquids according to the proportion, inoculating the three target bacteria liquids into the sterilized skim buffalo milk, wherein the inoculation amount is 20 percent of the total mass of the skim buffalo milk, and then placing the skim buffalo milk in a shaking table at 37 ℃ for 22 hours to obtain the composite strain.
The preparation method of the yoghourt comprises the following steps: heating skim buffalo milk to 75 ℃, then adding 9% of white granulated sugar and 0.7% of sodium glutamate, uniformly stirring, heating to 80-90 ℃, sterilizing for 10min, and cooling to 37 ℃ for later use; inoculating the composite strain into the skim buffalo milk treated by the method, wherein the inoculation amount is 8 percent of the total mass of the skim buffalo milk, fully stirring uniformly and then bottling; then placing the skimmed buffalo milk filled in the cup at the constant temperature of 40 ℃ for fermenting for 3-5h until the milk curd is formed, and regarding the fermentation end point when the pH value is 4.0-4.5 to obtain fermented yoghourt; and finally, putting the fermented yoghourt into a refrigerated cabinet with the temperature of 4 ℃, and after-ripening for 24 hours to obtain the finished product of the buffalo milk.
Example 5
The buffalo milk yoghourt rich in gamma-aminobutyric acid is set yoghourt. The yoghourt takes skim buffalo milk as a main material, and is prepared by adding 9% of white granulated sugar, 0.7% of sodium glutamate and 10% of composite strain in the total mass of the skim buffalo milk and fermenting. The skim buffalo milk is prepared by filtering and purifying qualified fresh buffalo milk and then performing milk fat separation, wherein the protein content is 6.12%, the fat content is 0.3%, and the acidity is less than or equal to 16 DEG T. The composite strain is prepared by activating lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 1: 4: 4, the specific activating preparation method comprises the following steps: respectively carrying out continuous secondary revival subculture on the freeze-dried lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus in an MRS slant culture medium to obtain strains; then respectively inoculating the strains into 100ml of MRS broth culture medium, and carrying out shake cultivation for 21h at 38 ℃ to obtain culture bacteria liquid; centrifuging the culture bacteria liquid for 9min at 4900rpm by a centrifuge, and collecting supernatant to obtain target bacteria liquid, namely activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus; and finally, uniformly mixing the three target bacteria liquids according to the proportion, inoculating the three target bacteria liquids into the sterilized skim buffalo milk, wherein the inoculation amount is 20 percent of the total mass of the skim buffalo milk, and then placing the skim buffalo milk in a shaking table at 37 ℃ for 24 hours to obtain the composite strain.
The preparation method of the yoghourt comprises the following steps: firstly heating skim buffalo milk to 75 ℃, then adding 9% of white granulated sugar and 0.7% of sodium glutamate, uniformly stirring, heating to 90 ℃, sterilizing for 10min, and cooling to 37 ℃ for later use; inoculating the composite strain into the treated skim buffalo milk, wherein the inoculation amount is 10% of the total mass of the skim buffalo milk, fully stirring uniformly and then bottling; then placing the skimmed buffalo milk filled in the cup at the constant temperature of 40 ℃ for fermenting for 3-5h until the milk curd is formed, and regarding the fermentation end point when the pH value is 4.0-4.5 to obtain fermented yoghourt; and finally, putting the fermented yoghourt into a refrigerated cabinet with the temperature of 4 ℃, and after-ripening for 24 hours to obtain the finished product of the buffalo milk.
Example 6
And (3) comparison test: the fermented buffalo milk yogurt rich in gamma-aminobutyric acid (GABA) of example 1, example 2, example 3, example 4 and example 5 is compared with two commercially available buffalo milk yogurt control groups 1 and 2 (the strains are mixed strains of traditional Lactobacillus bulgaricus and Streptococcus thermophilus), and the acidity, the viable count of lactic acid bacteria, the whey folding amount and the sensory evaluation condition of the yogurt after being preserved for 10 days at 4 ℃ are respectively detected, and the results are shown in the following table 1:
TABLE 1 analysis table of comparative test results
As can be seen from the above table 1, in the yoghurts obtained in the embodiments 1 to 5 of the present invention, the number of live bacteria of lactic acid bacteria is higher than that of the commercially available buffalo milk of the control group 1 and the control group 2; whey out was less than both of the commercial control products; the acidity of the yoghurt is lower after the yoghurt is preserved for 10 days at 4 ℃, which indicates that the product of the invention has controllable post-acidification degree, and the yoghurt can be effectively prolonged by adopting three bacteria of lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus which are separated from buffalo milk to mix and ferment the yoghurt; particularly, the GABA content in the commercial buffalo milk in the control group is zero, which indicates that the ordinary traditional yogurt strains do not produce GABA, but the yogurt obtained in the examples 1 to 5 of the invention contains GABA, wherein the GABA yield in the example 2 is the highest and reaches 3.6 g/kg; in addition, the products obtained in the embodiments 1 to 5 of the invention have good sensory evaluation, and the physical, chemical and microbial quality indexes of the products meet the current national standard requirements.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (7)
1. The buffalo yoghurt rich in gamma-aminobutyric acid is characterized in that skim buffalo milk is used as a main material, white granulated sugar accounting for 8% of the total mass of the skim buffalo milk, sodium glutamate accounting for 0.6% of the total mass of the skim buffalo milk and a compound strain accounting for 7% of the total mass of the skim buffalo milk are added and fermented to prepare the yoghurt, the compound strain is formed by mixing activated lactococcus lactis subsp lactis, lactobacillus plantarum and streptococcus thermophilus, and the lactococcus lactis subsp lactis and the lactobacillus plantarum are obtained by separating from fresh buffalo milk;
the lactococcus lactis subspecies lactis is lactococcus lactis subspecies lactis 25#, and the preservation number is CCTCC NO: M2015700; the Lactobacillus plantarum is Lactobacillus plantarum (Lactobacillus plantarum) No. 12, and the preservation number is CCTCC NO: M2015699.
2. The gamma-aminobutyric acid enriched buffalo milk according to claim 1, wherein: the skim buffalo milk is prepared by filtering fresh buffalo milk serving as a raw material to remove milk and then performing cream separation; the mass percent of the protein in the defatted buffalo milk is 5.5-6.8%, the mass percent of the fat is 0.2-0.5%, and the acidity is less than or equal to 16 DEG T.
3. The gamma-aminobutyric acid enriched buffalo milk according to claim 1, wherein: the composite strain is prepared from activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 1-2: 1: 1-2, and mixing.
4. The gamma-aminobutyric acid enriched buffalo milk according to claim 3, wherein: the composite strain is prepared from activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus according to the mass ratio of 2: 1: 1 are mixed.
5. The gamma-aminobutyric acid enriched buffalo milk according to claim 3, wherein: the composite strain is prepared by activating lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus respectively and mixing the activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus in proportion; the specific activating preparation method comprises the following steps:
a) continuously and secondarily reviving and subculturing lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus in an MRS slant culture medium respectively to obtain strains;
b) respectively inoculating the strains obtained in the step a) into 100ml of MRS broth culture medium, and performing shake culture at 35-40 ℃ for 20-24h to obtain culture bacteria liquid;
c) centrifuging the culture bacterial liquid obtained in the step b) for 6-10min at the rotating speed of 4800-5200rpm by using a centrifuge respectively, and collecting supernatant to obtain target bacterial liquid, namely activated lactococcus lactis subspecies lactis, lactobacillus plantarum and streptococcus thermophilus;
d) and c) uniformly mixing the three target bacterial liquids obtained in the step c) according to the proportion, inoculating the three target bacterial liquids into the sterilized skim buffalo milk, wherein the inoculation amount is 20% of the total mass of the skim buffalo milk, and then placing the skim buffalo milk in a shaking table at 37 ℃ for culturing for 16-24h to obtain the composite bacterial strain.
6. The buffalo milk yoghurt enriched with gamma-aminobutyric acid according to claim 5, wherein the yoghurt is a set yoghurt.
7. A process for the preparation of a milk of buffalo milk enriched with gamma-aminobutyric acid according to any one of claims 1 to 6, wherein the process comprises the steps of:
1) heating skim buffalo milk to 60-80 ℃, adding white granulated sugar and sodium glutamate according to mass percentage, stirring uniformly, heating to 80-95 ℃, sterilizing for 10-12min, and cooling to 35-37 ℃ for later use;
2) inoculating the composite strain into the skim buffalo milk treated in the step 1), fully stirring uniformly and then bottling;
3) fermenting the skimmed buffalo milk filled in the step 2) at the constant temperature of 38-41 ℃ for 3-5h until curd is formed, and taking the pH value of the skimmed buffalo milk as the fermentation end point when the pH value is 4.0-4.5 to obtain fermented yoghourt;
4) putting the fermented yoghourt obtained in the step 3) into a refrigerated cabinet with the temperature of 4-6 ℃, and after-ripening for 22-24h to obtain the finished product of the buffalo milk.
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| CN110257447B (en) * | 2019-06-20 | 2021-11-09 | 卡士乳业(深圳)有限公司 | Method for improving GABA (gamma-aminobutyric acid) production capacity of lactococcus lactis subsp |
| CN113512514B (en) * | 2021-07-01 | 2023-05-26 | 杭州娃哈哈科技有限公司 | Lactococcus lactis with depression improving effect and application thereof |
| CN115380953B (en) * | 2022-08-31 | 2023-09-19 | 桂林理工大学 | Fermented milk with ACE (angiotensin converting enzyme) inhibitory activity and preparation method thereof |
| CN115530325B (en) * | 2022-10-20 | 2024-02-27 | 江南大学 | Chickpea flour rich in gamma-aminobutyric acid and application thereof |
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