CN115380953B - Fermented milk with ACE (angiotensin converting enzyme) inhibitory activity and preparation method thereof - Google Patents
Fermented milk with ACE (angiotensin converting enzyme) inhibitory activity and preparation method thereof Download PDFInfo
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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
- A23C9/123—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
- 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
- A23V2400/123—Bulgaricus
-
- 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
- A23V2400/249—Thermophilus
Abstract
The invention provides a preparation method of fermented milk with ACE inhibition activity, which comprises the following steps: (1) Adding concentrated milk protein into preheated buffalo milk, stirring uniformly, and homogenizing; (2) After sterilization, cooling to room temperature, inoculating lactobacillus plantarum with the preservation number of CGMCC No.19748 and a yoghurt starter; (3) And (5) preserving heat, fermenting, and cooling after the fermentation is completed to terminate the fermentation. The invention also provides fermented milk prepared by the preparation method. According to the fermented milk with ACE inhibition activity and the preparation method thereof, provided by the invention, the fermented milk product with high ACE inhibition rate is finally obtained through the selection of raw materials and fermentation strains, the optimization of the formula and the fermentation process, the specific requirements of consumers can be met to the greatest extent, and the fermented milk has a broad market prospect.
Description
Technical Field
The invention relates to the field of dairy products, in particular to fermented milk with ACE (angiotensin converting enzyme) inhibitory activity and a preparation method thereof.
Background
Hypertension is one of the most common cardiovascular diseases, and is a major risk factor for diseases such as stroke, myocardial infarction and other cardiovascular system injuries, and seriously threatens the health of human beings. Drugs are the primary method of treating hypertension, and Angiotensin Converting Enzyme (ACE) inhibitors are widely used in clinic as antihypertensive drugs. However, ACE inhibitors such as captopril are currently synthetic drugs, and long-term administration of ACE inhibitors is likely to cause side effects such as cough, loss of taste and impairment of renal function. Therefore, the research and development of food-derived antihypertensive products has important significance for human health.
Probiotics (Probiotics) are a type of living microbial preparation that can improve the host's microecological balance and benefit the host's health when taken in proper amounts. It has been found that certain probiotics not only can reduce blood pressure by improving intestinal flora, but also can hydrolyze milk proteins to release various ACE inhibitory peptides through a specific protease system, thereby exerting the antihypertensive effect of ACE inhibitors. At present, the fermented dairy product with the blood pressure reducing function is mainly prepared by adding functional components such as sweet tea extract (CN 111771976A), mixed enzymolysis liquid of red bean milk, pine pollen and sweet osmanthus (ZL 201510137934.4), calcium hydroxide (CN 110623068A), buffalo milk antihypertensive peptide (ZL 201610154801.2) and the like, and adding a fermenting agent (with or without probiotics). However, in the existing products, the inhibition activity of ACE is low, and the actual demands of consumers are difficult to meet.
In view of this, the present invention has been made.
Disclosure of Invention
The invention aims to provide fermented milk with ACE inhibition activity and a preparation method thereof, and the prepared fermented milk product has an ACE inhibition rate of more than 95% by improving raw materials, fermentation strains and fermentation technology.
The technical scheme adopted by the invention for achieving the purpose is as follows:
the invention provides a preparation method of fermented milk with ACE inhibition activity, which comprises the following steps:
(1) Adding concentrated milk protein into preheated buffalo milk, stirring uniformly, and homogenizing;
(2) Sterilizing, cooling to room temperature after sterilization, inoculating lactobacillus plantarum with the preservation number of CGMCC No.19748 and a yoghurt starter, uniformly mixing and canning;
(3) And (5) preserving heat, fermenting, and cooling after the fermentation is completed to terminate the fermentation.
Preferably or alternatively, the pre-heating temperature of buffalo milk in step (1) is from 90 to 100 ℃, preferably 95 ℃.
Preferably or alternatively, in the step (1), the amount of the concentrated milk protein added is 0.5 to 0.9% by mass of buffalo milk; preferably, the amount of the concentrated milk protein added is 0.5%.
Preferably or alternatively, in step (1), the homogenisation pressure is from 175 to 180Bar.
Preferably or alternatively, in step (2), the sterilization temperature is 93-97 ℃ and the sterilization time is 4-6min.
Preferably or alternatively, in the step (2), the inoculation amount of lactobacillus plantarum with the preservation number of CGMCC No.19748 is 2-4% of the volume of the buffalo milk; preferably, the lactobacillus plantarum is inoculated in an amount of 3%.
Lactobacillus plantarum with the preservation number of CGMCC No.19748 is preserved in China general microbiological culture Collection center (China center for type culture Collection) at the time of 4 months and 27 days in 2020: the strain is disclosed by the prior art at the institute of microbiology, national academy of sciences, 1 st, 3 rd, north Chen, west, of the Chaoyang area, beijing, china.
Preferably or alternatively, in the step (2), the yoghurt starter is a compound microbial inoculum of lactobacillus bulgaricus and streptococcus thermophilus, and the addition amount is 0.2-0.4 percent of the mass of the buffalo milk
Preferably or alternatively, in the step (3), the temperature of the insulation fermentation is 35-43 ℃; preferably, the incubation fermentation temperature is 39 ℃.
Preferably or alternatively, in the step (3), the time of heat preservation and fermentation is 8-10h; preferably, the incubation fermentation time is 10 hours.
On the other hand, the invention also provides fermented milk with ACE inhibitory activity, and the fermented milk is prepared by adopting the preparation method of the fermented milk with ACE inhibitory activity.
Advantageous effects
The invention provides fermented milk with ACE inhibition activity and a preparation method thereof, and the fermented milk product with the ACE inhibition rate exceeding 95% is prepared by selecting and improving raw materials, fermentation strains and fermentation process.
Drawings
FIG. 1 is a graph showing the results of ACE inhibition ratios of the products of examples 1 to 3 and comparative examples 1 to 2 measured in effect example 1;
FIG. 2 is a graph showing the results of ACE inhibition ratios of the products of examples 3 to 5 and comparative examples 3 to 4 measured in effect example 1;
FIG. 3 is a graph showing the results of ACE inhibition ratios of the products of examples 3, 6-7 and comparative examples 5-6 measured in effect example 1;
FIG. 4 is a graph showing the results of ACE inhibition ratios of examples 3 and 8 to 11 measured in effect example 1.
Detailed Description
The present invention will be described more fully hereinafter with reference to the accompanying drawings and preferred examples for the purpose of facilitating understanding of the present invention, but the scope of the present invention is not limited to the following specific examples.
Unless defined otherwise, all technical and scientific terms used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the scope of the present invention.
Unless otherwise specifically indicated, the various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or may be prepared by existing methods.
The compound microbial agents of lactobacillus bulgaricus and streptococcus thermophilus used in each example and comparative example in the present invention were purchased from danish company;
the buffalo milk used in each of the examples and comparative examples in the present invention was purchased from the company of the milk industry, inc. Of guangxi;
the common cow milk used in each of the examples and comparative examples in the present invention was purchased from Beijing ternary food Co.
Example 1
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748 accounting for 2% of the volume of buffalo milk, wherein the concentration of Lactobacillus plantarum in the seed solution is 2.39X10% 10 CFU/mL, lactobacillus bulgaricus and thermophilus with water buffalo milk quality of 0.3%A compound bacterial agent of streptococcus. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Example 2
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Example 3
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Comparative example 1
The comparative example provides a fermented milk with ACE inhibitory activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748 1% of the volume of buffalo milk, wherein the concentration of Lactobacillus plantarum in the seed solution is 2.39X10% 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Comparative example 2
The comparative example provides a fermented milk with ACE inhibitory activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Example 4
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.7% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Example 5
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.9% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Comparative example 3
The comparative example provides a fermented milk with ACE inhibitory activity, which is prepared by the following steps.
Adding 0.1% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Comparative example 4
The comparative example provides a fermented milk with ACE inhibitory activity, which is prepared by the following steps.
Adding 0.3% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
At 95 DEG CSterilizing for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10% 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Example 6
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 9 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Example 7
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Comparative example 5
The comparative example provides a fermented milk with ACE inhibitory activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 7 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Comparative example 6
The comparative example provides a fermented milk with ACE inhibitory activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 37deg.C for 11 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibiting activity.
Example 8
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating Buffalo milk with preservation number of CGMCC 4% by volumeSeed solution of Lactobacillus plantarum No.19748, wherein the concentration of Lactobacillus plantarum in the seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 39deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Example 9
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 41 deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibiting activity.
Example 10
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 43deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibiting activity.
Example 11
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing at 175 Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 35deg.C for 8 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibiting activity.
Effect example 1
In this effect example, spectrophotometry was used to determine ACE inhibition rate.
Taking 200 mu L of 0.88mM furan acrylyl tripeptide (FAPCG), preheating for 5min at 37 ℃, sequentially adding 20 mu L of whey and 20 mu L of ACE after centrifugation of each sample, uniformly mixing, immediately measuring the initial absorbance a1 at 340nm, reacting for 15min at 37 ℃, and measuring the absorbance a2 at 340nm again; meanwhile, double distilled water is adopted as a blank control group, the absorbance at 340nm before the reaction is measured as b1, and the absorbance at 340nm after the reaction is measured as b2.
And calculates the ACE inhibition ratio according to the formula ACE inhibition ratio (%) = (b 1-b 2) - (a 1-a 2)/(b 1-b 2) ×100%.
ACE inhibition rates of the products of examples 1 to 3 and comparative examples 1 to 2 were measured, respectively, and the results are shown in fig. 1.
The ACE inhibition rates of the products of examples 3 to 5 and comparative examples 3 to 4 were measured, respectively, and the results are shown in FIG. 2.
ACE inhibition rates of the products of examples 3, 6 to 7 and comparative examples 5 to 6 were measured, respectively, and the results are shown in fig. 3.
ACE inhibition rates of examples 3 and 8 to 11 were measured, and the results are shown in fig. 4.
From figures 1-4, it can be seen that when the added amount of the concentrated milk protein is 0.5-0.9%, the inoculation amount of lactobacillus plantarum with the preservation number of CGMCC No.19748 is 2-4%, the fermentation time is 8-10h, and the ACE inhibition rate of the product is higher when the fermentation temperature is 35-43 ℃ compared with other conditions.
Example 12
The parameters were further optimized for the results of effect example 1.
The embodiment provides fermented milk with ACE inhibition activity, which is prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product with ACE inhibitory activity.
Comparative example 7
The comparative example provides a fermented milk prepared by the following steps.
The buffalo milk preheated to 90-100 ℃ is homogenized at 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of buffalo milk. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product.
Comparative example 8
The comparative example provides a fermented milk prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of buffalo milk into buffalo milk preheated to 90-100deg.C, mixing, and homogenizing under 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, and inoculating lactobacillus bulgaricus and Streptococcus thermophilus compound microbial inoculum with mass of 0.3% of that of buffalo milk. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product.
Comparative example 9
The comparative example provides a fermented milk prepared by the following steps.
The buffalo milk preheated to 90-100 ℃ is homogenized at 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, and inoculating lactobacillus bulgaricus and Streptococcus thermophilus compound microbial inoculum with mass of 0.3% of that of buffalo milk. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product.
Comparative example 10
The comparative example provides a fermented milk prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of common cow milk into common cow milk preheated to 90-100deg.C, mixing, and homogenizing under 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and 0.3% of the mass of milk. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product.
Comparative example 11
The comparative example provides a fermented milk prepared by the following steps.
Homogenizing common cow milk preheated to 90-100deg.C under 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, inoculating seed solution of Lactobacillus plantarum with preservation number of CGMCC No.19748, wherein the concentration of Lactobacillus plantarum in seed solution is 2.39X10 10 CFU/mL, and cowA compound microbial inoculum of Lactobacillus bulgaricus and Streptococcus thermophilus with milk quality of 0.3%. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product.
Comparative example 12
The comparative example provides a fermented milk prepared by the following steps.
Adding 0.5% concentrated milk protein by weight of common cow milk into common cow milk preheated to 90-100deg.C, mixing, and homogenizing under 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, and inoculating lactobacillus bulgaricus and Streptococcus thermophilus compound microbial inoculum with mass of 0.3% of cow milk. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product.
Comparative example 13
The comparative example provides a fermented milk prepared by the following steps.
Homogenizing common cow milk preheated to 90-100deg.C under 180Bar.
Sterilizing at 95deg.C for 5min, cooling to room temperature, and inoculating lactobacillus bulgaricus and Streptococcus thermophilus compound microbial inoculum with mass of 0.3% of cow milk. Mixing well.
Fermenting at 39deg.C for 10 hr, and refrigerating to stop fermentation to obtain fermented milk product.
Effect example 2
The ACE inhibition rates of example 12 and comparative examples 7 to 13 were measured by the method for measuring ACE inhibition rate described in effect example 1, and the results are shown in table 1.
Table 1 results of various examples and comparative examples table of ACE inhibition rates of fermented milk
As can be seen from Table 1, the ACE inhibition rate of the final product is far higher than that of other similar products by the selection of raw materials and fermentation strains and the improvement of the formula.
Effect example 3
The polypeptide content and distribution of the fermented milk of example 12 and comparative example 10 were qualitatively determined by LC-MS/MS, respectively, by the following method: separating the supernatant of the sample by adopting an ultrafiltration membrane with the molecular weight cut-off of 10k and 3k in sequence to obtain a polypeptide component II (3-10 k) and a component III (less than 3 k), further extracting small peptide by adopting an extracting solution (methanol: chloroform: water=3:1:4, V/V/V), and detecting by adopting LC-MS/MS after vacuum drying; the test results are shown in Table 2.
TABLE 2 Table of polypeptide profiles for fermented milk products of example 13 and comparative example 10
Table 2 shows the ACE inhibitory peptide content of the milk fermented from buffalo and common cow milk containing Lactobacillus plantarum B. As can be seen from Table 2, 96 amino acid sequences related to ACE inhibition are generated in the fermentation process of buffalo milk, and the quantity and the abundance of the amino acid sequences respectively account for 17.6% and 40.0% of total peptides, which are obviously higher than those of fermented milk produced by taking common cow milk as a raw material.
The invention provides the fermented milk with ACE inhibition activity and the preparation method thereof, and the fermented milk product with high ACE inhibition rate is finally obtained through the selection of raw materials and fermentation strains, the optimization of a formula and a fermentation process, so that the specific requirements of consumers can be met to the greatest extent, and the fermented milk has a wide market prospect.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (12)
1. A method for preparing fermented milk with ACE inhibitory activity, which is characterized by comprising the following steps:
(1) Adding concentrated milk protein into preheated buffalo milk, stirring uniformly, and homogenizing; the addition amount of the concentrated milk protein is 0.5-0.9% based on the mass of buffalo milk;
(2) Sterilizing, cooling to room temperature after sterilization, inoculating lactobacillus plantarum with the preservation number of CGMCC No.19748 and a yoghurt starter, uniformly mixing and canning; the inoculation amount of lactobacillus plantarum is 2-4% of the volume of buffalo milk, and the concentration of the bacterial liquid of lactobacillus plantarum is 2.39X10 10 CFU/mL；
(3) Preserving heat and fermenting for 8-10h, cooling and terminating fermentation after fermentation is completed, thus obtaining the final product.
2. The method for producing fermented milk with ACE inhibiting activity according to claim 1, wherein the preheating temperature of buffalo milk in step (1) is 90 to 100 ℃.
3. The method for producing fermented milk with ACE inhibiting activity according to claim 2, wherein the preheating temperature of buffalo milk in step (1) is 95 ℃.
4. The method for producing fermented milk having ACE inhibitory activity according to claim 1, wherein the amount of the concentrated milk protein added in step (1) is 0.5%.
5. The method for producing fermented milk with ACE inhibitory activity according to claim 1, wherein in step (1), the homogenizing pressure is 175 to 180Bar.
6. The method for producing fermented milk with ACE inhibitory activity according to claim 1, wherein in step (2), the sterilization temperature is 93 to 97 ℃ and the sterilization time is 4 to 6min.
7. The method for producing fermented milk with ACE inhibitory activity according to claim 1, wherein the inoculation amount of lactobacillus plantarum in step (2) is 3%.
8. The method for producing fermented milk with ACE inhibitory activity according to claim 1, wherein in the step (2), the yogurt starter is a compound microbial inoculum of lactobacillus bulgaricus and streptococcus thermophilus, and the addition amount is 0.2 to 0.4% based on the mass of buffalo milk.
9. The method for producing fermented milk with ACE inhibitory activity according to claim 1, wherein in the step (3), the temperature of the incubation fermentation is 35 to 43 ℃.
10. The method for producing fermented milk with ACE inhibitory activity according to claim 9, wherein in the step (3), the temperature of the incubation fermentation is 39 ℃.
11. The method for producing fermented milk with ACE inhibitory activity according to claim 1, wherein in step (3), the incubation fermentation time is 10 hours.
12. A fermented milk with ACE inhibitory activity, characterized in that it is prepared by the method for preparing a fermented milk with ACE inhibitory activity according to any one of claims 1 to 11.
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