CN111034730B - Method for cultivating bean sprouts rich in GABA and anthocyanin and method for preparing fermented milk beverage by using bean sprouts - Google Patents
Method for cultivating bean sprouts rich in GABA and anthocyanin and method for preparing fermented milk beverage by using bean sprouts Download PDFInfo
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- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 title claims abstract description 143
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N35/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical
- A01N35/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having two bonds to hetero atoms with at the most one bond to halogen, e.g. aldehyde radical containing aliphatically bound aldehyde or keto groups, or thio analogues thereof; Derivatives thereof, e.g. acetals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
-
- 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
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/133—Fruit or vegetables
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Environmental Sciences (AREA)
- Wood Science & Technology (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Pest Control & Pesticides (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Non-Alcoholic Beverages (AREA)
Abstract
The invention discloses a method for cultivating sprouts rich in GABA and anthocyanin and a method for preparing fermented milk beverage thereof, which are characterized in that the cultivating method comprises the steps of soaking Chinese violet seeds in sodium hypochlorite solution, washing the Chinese violet seeds with pure water, soaking and draining the Chinese violet seeds in a mixed solution containing 2.7-10.8g/L glucose and 0.5-1.5 g/L sedum aizoon total flavonoids, then replacing soaking liquid every day after 12/12h dark light treatment, cleaning the sprouts, draining, freeze drying and crushing the sprouts to obtain the sprouts rich in GABA and anthocyanin; the preparation method of the fermented milk beverage comprises the following steps: the bud vegetable powder rich in GABA and anthocyanin and the sugar-containing milk beverage prepared from wheat kernels and corn nibs are mixed in equal volume and boiled, filtered after ultrasonic treatment, the filtrate is added with lactobacillus plantarum suspension, and the mixture is canned, kept warm, fermented and ripened to obtain the fermented milk beverage rich in GABA and anthocyanin.
Description
Technical Field
The invention relates to a preparation method of a fermented milk beverage, in particular to a cultivation method of sprouts rich in GABA and anthocyanin and a preparation method of the fermented milk beverage.
Background
Gamma-aminobutyric acid (GABA) is a ubiquitous non-protein amino acid compound, and is widely present in animals and plants. GABA acts as an inhibitory neurotransmitter in the brain and spinal cord of mammals, regulating blood pressure and heart rate, relieving pain and anxiety, and thus is commonly used as an antihypertensive agent, diuretic, sedative and a neurotransmitter that inhibits sympathetic nerve function. Recent studies have shown that GABA is also a potent agent for insulin secretion by the pancreas, and is effective in preventing diabetes. Owing to these physiological functions, GABA has been approved as a new resource food by the national ministry of health in 2009. However, with the increase of age and pressure, the production amount of GABA in human body is gradually reduced, so that the supplement of GABA from an external source has important significance to the health of human body.
Currently, GABA products from plant sources are concerned about due to the characteristics of safety and naturalness, but the GABA products are generally low in many plants and generally between 0.3 and 32.5 mu mol/g. In response to external biotic and abiotic stress, the GABA branch of plants rapidly increases and accumulates GABA in large amounts. In 1996, rice germ products rich in GABA were commercialized in Japan. In 2001, functional food ingredients rich in GABA, which are developed and manufactured by Japanese scientists using rice germ and other raw materials, have been widely used in products such as beverages, jams, cakes, biscuits, seasonings and the like. Recently, the research and development of GABA food by Chinese scientists have entered into commercialization, and more functional products rich in GABA are developed and put on the market. The yogurt has high nutritional value and can further enrich GABA through strain fermentation, but the GABA content is low through strain fermentation.
Disclosure of Invention
The invention aims to provide a method for cultivating sprouts rich in GABA and anthocyanin and high in gamma-aminobutyric acid and anthocyanin content and a method for preparing fermented milk beverage by using the sprouts.
The technical scheme adopted by the invention for solving the technical problems is as follows:
1. a method for cultivating sprouts rich in GABA and anthocyanin comprises the following steps: soaking seeds of the purple Chinese violet in a sodium hypochlorite solution of 12-18wt% for 3-5 hours, removing suspended particles, washing with pure water until the pH value is neutral, soaking in a mixed solution of 2.7-10.8g/L glucose and 0.5-1.5 g/L total flavonoids of the sedum aizoon at 28-32 ℃ for 22-26 hours, replacing the soaking solution every 5-7 hours, draining, placing on a gauze bed of a seed machine, then continuously soaking with the mixed solution for germination under the conditions of 12-hour illumination and 12-hour dark treatment, replacing the soaking solution every day, taking out the roots after 7 days, cleaning the sprouts, and draining to obtain the sprouts rich in GABA and anthocyanin.
Preferably, the soaking solution is a mixed solution containing 5.4g/L glucose and 1.0g/L herba Sedi Aizoon total flavonoids.
The preparation method of the sedum aizoon total flavonoids comprises the following specific steps:
(1) separating fresh picked herba Sedi Aizoon stems and leaves, respectively drying in 60 deg.C oven, pulverizing with pulverizer, sieving with 60 mesh sieve, collecting herba Sedi Aizoon powder 1000 g, adding 40L 70% ethanol solution, homogenizing, ultrasonic extracting at 60 deg.C for 60 min, leaching at 60 deg.C for 2 hr, and filtering to obtain supernatant;
(2) concentrating the supernatant obtained in the step (1) by a rotary evaporator under reduced pressure, recovering the solvent, heating to 45-50 ℃, and adding absolute ethyl alcohol with volume twice that of the concentrated solution after the concentration is finished;
(3) treating the solution obtained in the step (2) at a low temperature of 4 ℃ for 24 hours, and performing suction filtration to obtain a supernatant;
(4) concentrating the supernatant obtained in the step (3) through a rotary evaporator under reduced pressure, recovering the solvent, and heating to 45-50 ℃; adding the concentrated solution into a 20% ethanol solution, diluting the concentrated solution into a sedum aizoon extract solution with the flavone concentration of 2.5 mg/mL, then adding macroporous resin which is treated by acid, alkali and alcohol until the solution is completely swelled, statically adsorbing for 24 hours, filtering, discarding filtrate, adding 80% ethanol solution with the same volume as the 20% ethanol solution for analysis, and filtering to obtain supernatant after the analysis is finished, wherein the ratio of the sedum aizoon extract solution to the macroporous resin is 10 mL: 1g of a compound;
(5) and (4) carrying out reduced pressure concentration on the supernatant obtained in the step (4) by using a rotary evaporator, recovering the solvent, heating to 45-50 ℃, obtaining a concentrated solution, namely the herba sedi aizoon total flavone, and freeze-drying for later use.
2. A preparation method of fermented milk beverage rich in GABA and anthocyanin comprises the following steps:
(1) preparation method of bean sprouts rich in GABA and anthocyanin
Soaking seeds of the purple Chinese violet in a sodium hypochlorite solution of 12-18wt% for 3-5 hours, removing suspended particles, washing with pure water until the pH value is neutral, soaking in a mixed solution of 2.7-10.8g/L glucose and 0.5-1.5 g/L total flavonoids of the sedum aizoon at 28-32 ℃ for 22-26 hours, draining, placing on a gauze bed of a seed machine, continuously soaking with the mixed solution for germination under the conditions of 12-hour illumination and 12-hour dark treatment, replacing the soaking solution every day, taking out the roots after 7 days, cleaning the sprouts, draining, placing in a freeze dryer for 22-26 hours, and crushing to obtain sprout powder rich in GABA and anthocyanin;
(2) preparation of sugar-containing milk beverage
Soaking wheat grains in 28-32 deg.C water for 22-26 hr, germinating for 4 days, taking out, and cutting; soaking the corn particles in water for 3-5 hours, steaming at high temperature, cooling to 40-50 ℃, and taking out the corn particles; then mixing and fermenting the wheat crumbs and the corn crumbs according to the volume ratio of 1:10 for 5-6 hours, squeezing, concentrating to 1/5 of the original volume, and fixing the volume to 10 times of the volume of the concentrated solution by using pure milk to obtain the sugar-containing milk beverage;
(3) preparing a lactobacillus plantarum suspension:
inoculating lactobacillus plantarum into an MRS culture medium, and performing shake cultivation for 22-26 hours at 36-38 ℃ to obtain lactobacillus plantarum suspension;
(4) and (3) mixing the bud vegetable powder rich in GABA and anthocyanin prepared in the step (1) and the sugar-containing milk beverage prepared in the step (2) according to the equal volume, boiling, sealing, carrying out ultrasonic treatment at the temperature of 18-22 ℃ for 1-3 hours, filtering, adding the filtrate into the lactobacillus plantarum suspension prepared in the step (3), canning, preserving the heat at the temperature of 40-44 ℃ for 3-5 hours, and carrying out after-ripening at the temperature of 0-4 ℃ for 14-18 hours to obtain the fermented milk beverage rich in GABA and anthocyanin.
Preferably, the soaking solution is a mixed solution containing 5.4g/L glucose and 1.0g/L herba Sedi Aizoon total flavonoids.
Compared with the prior art, the invention has the advantages that:
1. by adopting the preparation process, because the mixed solution culture of glucose and the total flavonoids of sedum aizoon is adopted in the germination process of the purple-flower sprouts, the tricarboxylic acid cycle is influenced by utilizing the glucose to enrich acetyl coenzyme A through a glucose metabolism way so as to influence GABA branches, and the total flavonoids of sedum aizoon influences alpha-ketoglutaric acid in the tricarboxylic acid cycle by producing a large amount of calcium ions and cooperating with the glucose, and simultaneously influences the activity of GAD. Therefore, the GABA content of the fermented milk beverage is far higher than that of functional yogurt produced by the prior art, and the fermented milk beverage has higher nutritional value.
2. The purple cauliflower is a variety of cauliflower, other characters of the purple cauliflower are similar to those of the cauliflower, but the purple cauliflower is rich in anthocyanin, so that the yogurt made of the purple cauliflower has a clear color, and the anthocyanin content of the yogurt is far higher than that of ordinary yogurt.
3. The invention firstly proposes that the flavonoids are used for culturing the plant sprouts, and experiments show that the GABA content of the sprouts cultured after the flavonoids are added is 1 time higher than that of a control.
4. The maltose prepared from the wheat and corn particles is a healthier traditional sugar, does not bring too much burden to human body, and milk beverage is required to be fermented when preparing yoghourt, so that the sugar-containing milk beverage is prepared in advance, a little astringent taste brought by adding bean sprouts can be covered, and the integration of mouthfeel in the fermentation process is facilitated.
Drawings
FIG. 1 is a graph showing the effect of immersion treatment of glucose solutions of different concentrations on the change of GABA content, with the abscissa representing the concentration of the glucose solution of different concentrations and the ordinate representing the GABA content;
FIG. 2 shows the effect of soaking glucose solutions of different concentrations on the change of anthocyanin content, with the abscissa representing the concentration of the glucose solution and the ordinate representing the anthocyanin content;
FIG. 3 is a graph of the effect of treatment of various concentrations of Philippine violet herb total flavonoids on the change in GABA content, with the abscissa representing various concentrations of Philippine violet herb total flavonoids and the ordinate representing GABA content;
FIG. 4 shows the effect of treatment of Philippine fleabane total flavone solutions of different concentrations on the change of anthocyanin content, with the abscissa representing the concentration of Philippine fleabane total flavone solution and the ordinate representing the anthocyanin content.
Detailed Description
The invention is described in further detail below with reference to the accompanying examples.
Detailed description of the preferred embodiment
A method for cultivating sprouts rich in GABA and anthocyanin comprises the following steps: soaking seeds of the purple Chinese violet in a sodium hypochlorite solution of 12-18wt% for 3-5 hours, removing suspended particles, washing with pure water until the pH value is neutral, soaking in a mixed solution of 2.7-10.8g/L glucose and 0.5-1.5 g/L total flavonoids of the sedum aizoon at 28-32 ℃ for 22-26 hours, replacing the soaking solution every 5-7 hours, draining, placing on a gauze bed of a seed machine, then continuously soaking with the mixed solution for germination under the conditions of 12-hour illumination and 12-hour dark treatment, replacing the soaking solution every day, taking out the roots after 7 days, cleaning the sprouts, and draining to obtain the sprouts rich in GABA and anthocyanin. The preferred soaking solution is a mixed solution containing 5.4g/L glucose and 1.0g/L herba Sedi Aizoon total flavonoids.
The preparation method of the herba sedi aizoon total flavonoids comprises the following specific steps:
(1) separating fresh picked herba Sedi Aizoon stems and leaves, respectively drying in 60 deg.C oven, pulverizing with pulverizer, sieving with 60 mesh sieve, collecting herba Sedi Aizoon powder 1000 g, adding 40L 70% ethanol solution, homogenizing, ultrasonic extracting at 60 deg.C for 60 min, leaching at 60 deg.C for 2 hr, and filtering to obtain supernatant;
(2) concentrating the supernatant obtained in the step (1) by a rotary evaporator under reduced pressure, recovering the solvent, heating to 45-50 ℃, and adding absolute ethyl alcohol with volume twice that of the concentrated solution after the concentration is finished;
(3) treating the solution obtained in the step (2) at a low temperature of 4 ℃ for 24 hours, and performing suction filtration to obtain a supernatant;
(4) concentrating the supernatant obtained in the step (3) through a rotary evaporator under reduced pressure, recovering the solvent, and heating to 45-50 ℃; adding the concentrated solution into a 20% ethanol solution, diluting the concentrated solution into a sedum aizoon extract solution with the flavone concentration of 2.5 mg/mL, then adding macroporous resin which is treated by acid, alkali and alcohol until the solution is completely swelled, statically adsorbing for 24 hours, filtering, discarding filtrate, adding 80% ethanol solution with the same volume as the 20% ethanol solution for analysis, and filtering to obtain supernatant after the analysis is finished, wherein the ratio of the sedum aizoon extract solution to the macroporous resin is 10 mL: 1g of a compound;
(5) and (4) carrying out reduced pressure concentration on the supernatant obtained in the step (4) by using a rotary evaporator, recovering the solvent, heating to 45-50 ℃, obtaining a concentrated solution, namely the herba sedi aizoon total flavone, and freeze-drying for later use.
Detailed description of the invention
A preparation method of fermented milk beverage rich in GABA and anthocyanin comprises the following steps:
(1) preparation method of bean sprouts rich in GABA and anthocyanin
Soaking Chinese violet seeds serving as a raw material in a sodium hypochlorite solution of 12-18wt% for 3-5 hours, removing suspended particles, washing with pure water until the pH value is neutral, soaking in a mixed solution containing 5.4g/L glucose and 1.0g/L sedum aizoon total flavonoids at 28-32 ℃ for 22-26 hours, draining, placing on a gauze bed of a seed machine, continuously soaking with the mixed solution for germination under the conditions of 12-hour illumination and 12-hour dark treatment, replacing the soaking solution every day, taking out roots on the 7 th day, cleaning sprouts, draining, placing in a freeze dryer for 22-26 hours, and crushing to obtain sprout powder rich in GABA and anthocyanin;
(2) preparation of sugar-containing milk beverage
Soaking wheat grains in 28-32 deg.C water 2 times the volume of wheat grains for 22-26 hr, germinating for 4 days, and cutting; soaking the corn particles for 3-5 hours, and cooling after high-temperature cooking; mixing and fermenting the wheat crumbs and the corn crumbs for 5-6 hours according to the volume ratio of 1:10, squeezing, concentrating to 1/5 of the original volume, and fixing the volume to 10 times of the volume of the concentrated solution by using pure milk to obtain the sugar-containing milk beverage;
(3) preparation of Lactobacillus plantarum suspensions
Inoculating lactobacillus plantarum into an MRS culture medium, and performing shake cultivation for 22-26 hours at 36-38 ℃ to obtain lactobacillus plantarum suspension;
(4) and (3) mixing the bud vegetable powder rich in GABA and anthocyanin prepared in the step (1) and the sugar-containing milk beverage prepared in the step (2) according to the equal volume, boiling, sealing, carrying out ultrasonic treatment at the temperature of 18-22 ℃ for 1-3 hours, filtering, adding the filtrate into the lactobacillus plantarum suspension prepared in the step (3), canning, preserving heat at the temperature of 40-44 ℃ for 3-5 hours, and carrying out after-ripening at the temperature of 0-4 ℃ for 14-18 hours to obtain the fermented milk beverage rich in GABA and anthocyanin.
Comparative test
1. Effect of glucose treatment at various concentrations on GABA content and anthocyanins in Brassica oleracea
The method for culturing the sprouts rich in GABA and anthocyanin is the same as the step (1) in the embodiment, and is characterized in that the sprouts are cultured by glucose solutions with the concentrations of 0g/L, 2.7g/L, 5.4g/L, 10.8g/L, 16.2 g/L, 21.6 g/L and 27 g/L respectively, and the GABA content of the purple-flower sprouts at all concentrations is measured. As shown in the graph 1, the GABA enrichment effect in the purple-flower buds is the best when the concentration of the glucose solution is 5.4g/L, the GABA content is 2.07 mg/g, the concentration of the glucose solution is 10.8g/L, the GABA content is 1.53 mg/g, and the GABA content is 1.36 mg/g when the concentration of the glucose solution is 2.7g/L, so that the difference is large. As can be seen from FIG. 2, when the concentration of the glucose solution is 10.8g/L, the anthocyanin enrichment effect in the purple-flower sprouts is the best, the anthocyanin content is 6.8 mg/100g, when the concentration of the glucose solution is 5.4g/L, the anthocyanin enrichment effect in the purple-flower sprouts is the second best, and the anthocyanin content is 6.6 mg/100g, which are relatively close to each other. Therefore, the concentration of the glucose solution is preferably 5.4 g/L.
The method for measuring GABA content in the purple cauliflower sprouts comprises the following steps: weighing 0.4g of purple-flower bud vegetable sample, adding 2 mL of 0.05 mol/L lanthanum chloride solution, grinding in ice bath, oscillating at room temperature for 15 min, freezing at high speed (10000 r/min) (4 ℃) and centrifuging for 5min to obtain supernatant. Taking out 1.6 mL of supernatant, adding 320 mu L and 1 mol/L of potassium hydroxide solution, oscillating for 5min at room temperature, after complete precipitation, centrifuging for 5min at 10000 r/min and 4 ℃ to obtain GABA extracting solution, taking out 400 mu L of GABA extracting solution, adding 600 mu L, pH 9.0.0 of boric acid buffer solution, shaking up, sequentially adding 2 mL and 6% of phenol solution and 1mL and 5% of sodium hypochlorite solution, mixing, placing into a boiling water bath for reaction for 5min, placing into an ice bath for cooling for 5min after the color becomes blue-green, finally adding 2 mL and 60% of ethanol solution for stopping the reaction, and determining the value at 645nm wavelength.
The method for measuring the anthocyanin content in the purple cauliflower sprouts comprises the following steps: grinding 0.2g of herba Violae Japonicae sample in 1mL of methanol containing 1% (V/V) hydrochloric acid, freezing at high speed (13000 r/min), centrifuging at 4 deg.C for 15 min to obtain supernatant, and measuring absorbance at 535 nm. The anthocyanin content was calculated using the following formula:
2. Effect of treatment with flavones of varying concentrations on GABA and anthocyanin levels in Brassica oleracea
The method for culturing the sprouts rich in GABA and anthocyanin is the same as the step (1) of the specific embodiment, the optimal treatment concentration of the total flavonoids of the sedum aizoon is screened out on the basis of the screened optimal glucose concentration so as to obtain the optimal combination effect, and the difference is that the total flavonoids of the sedum aizoon with the concentrations of 0g/L, 0.5 g/L, 1.0g/L, 1.5 g/L, 2.0 g/L and 2.5 g/L are respectively mixed with 5.4g/L of glucose for culture, and the GABA and anthocyanin content in the brassica oleracea sprouts under each concentration is measured. The result is shown in fig. 3, and the comparison result of the GABA content in the broccoli sprouts at each concentration shows that when the concentration of the Philippine flemingia root total flavone solution is 1.0g/L, the GABA enrichment effect in the purple flower sprouts is the best, and the GABA content is 4.6 mg/g. As can be seen from FIG. 3, the purple-flower sprouts cultured with different concentrations of the total flavonoids of sedum aizoon can obtain more obvious GABA enrichment effect than the single glucose treatment under the determined preferred glucose concentration. It can also be seen that the most obvious effect is obtained when the total flavone content of herba Fimbristylis Dichotomae is 1.0g/L and when the extract is mixed with 5.4g/L glucose solution.
As can be seen from fig. 4, when the addition amount of the sedum aizoon total flavonoids is less than 1.0g/L, the anthocyanin content does not significantly change, but the anthocyanin content is increased from 6.6 mg/100g (no sedum aizoon total flavonoids are added) of the control group to 6.9 mg/100g when the addition amount of the sedum aizoon total flavonoids is 1.0g/L, and the anthocyanin synthesis is adversely affected as the treatment concentration of the sedum aizoon total flavonoids is increased. Therefore, the purple-flower bud vegetable cultured by mixing the 1.0g/L herba sedi aizoon total flavonoids and the 5.4g/L glucose solution is more suitable for preparing the yoghourt.
3. Comparison of GABA and anthocyanin contents in fermented milk beverage and conventional fermented milk beverage
Respectively taking 3mL of the conventional fermented milk beverage sold on the market and 3mL of the fermented milk beverage added with the broccoli sprouts, centrifuging for 10min at 4000g, and taking the supernatant to measure the GABA content and the anthocyanin content according to the method. Meanwhile, the acidity, viscosity, water holding rate and other relevant indexes are measured by taking appropriate amount.
Remarking: the preparation method of the fermented milk beverage obtained by soaking sprout with single 5.4g/L glucose is different from the preparation method of the fermented milk beverage of the invention in that: 5.4g/L glucose is adopted as the soak solution in the step (1); the preparation method of the fermented milk beverage obtained by soaking bean sprouts with 1.0g/L single sedum aizoon total flavonoids is the same as the preparation method of the fermented milk beverage provided by the invention, and the difference is that: the soaking solution in the step (1) adopts 1.0g/L of herba sedi aizoon total flavone solution.
The synergetic principle is as follows: the single glucose treatment can influence the content of endogenous sugar through exogenous sugar, influence tricarboxylic acid cycle, and then influence GABA branch road enrichment GABA. The single treatment of the total flavonoids of sedum aizoon affects the enrichment of calcium ions, and further affects the enrichment of alpha-ketoglutaric acid in tricarboxylic acid cycle, and simultaneously affects the activity of GAD. The two are mixed, and the key product of alpha-ketoglutaric acid can reach higher concentration through the promoting effect of the exogenous sugar on acetyl coenzyme A and the enriching effect of flavone on calcium ions, so that tricarboxylic acid cycle is promoted, and the GABA is enriched at higher concentration.
The above description is not intended to limit the present invention, and the present invention is not limited to the above examples. Those skilled in the art should also realize that changes, modifications, additions and substitutions can be made without departing from the true spirit and scope of the invention.
Claims (5)
1. The method for cultivating the bean sprouts rich in GABA and anthocyanin is characterized by comprising the following steps of: soaking seeds of the purple Chinese violet in a sodium hypochlorite solution of 12-18wt% for 3-5 hours, removing suspended particles, washing with pure water until the pH value is neutral, soaking in a mixed solution of 2.7-10.8g/L glucose and 0.5-1.5 g/L total flavonoids of the sedum aizoon at 28-32 ℃ for 22-26 hours, replacing the soaking solution every 5-7 hours, draining, placing on a gauze bed of a seed machine, then continuously soaking with the mixed solution for germination under the conditions of 12-hour illumination and 12-hour dark treatment, replacing the soaking solution every day, taking out the roots after 7 days, cleaning the sprouts, and draining to obtain the sprouts rich in GABA and anthocyanin.
2. The method for cultivating sprouts rich in GABA and anthocyanin according to claim 1, wherein the method comprises the following steps: the soak solution is a mixed solution containing 5.4g/L glucose and 1.0g/L herba Sedi total flavonoids.
3. The method for cultivating sprouts rich in GABA and anthocyanin according to claim 1, wherein the method for preparing the total flavonoids of sedum aizoon comprises the following specific steps:
(1) separating fresh picked herba Sedi Aizoon stems and leaves, respectively drying in 60 deg.C oven, pulverizing with pulverizer, sieving with 60 mesh sieve, collecting herba Sedi Aizoon powder 1000 g, adding 40L 70% ethanol solution, homogenizing, ultrasonic extracting at 60 deg.C for 60 min, leaching at 60 deg.C for 2 hr, and filtering to obtain supernatant;
(2) concentrating the supernatant obtained in the step (1) by a rotary evaporator under reduced pressure, recovering the solvent, heating to 45-50 ℃, and adding absolute ethyl alcohol with volume twice that of the concentrated solution after the concentration is finished;
(3) treating the solution obtained in the step (2) at a low temperature of 4 ℃ for 24 hours, and performing suction filtration to obtain a supernatant;
(4) concentrating the supernatant obtained in the step (3) through a rotary evaporator under reduced pressure, recovering the solvent, and heating to 45-50 ℃; adding the concentrated solution into a 20% ethanol solution, diluting the concentrated solution into a sedum aizoon extract solution with the flavone concentration of 2.5 mg/mL, then adding macroporous resin which is treated by acid, alkali and alcohol until the solution is completely swelled, statically adsorbing for 24 hours, filtering, discarding filtrate, adding 80% ethanol solution with the same volume as the 20% ethanol solution for analysis, and filtering to obtain supernatant after the analysis is finished, wherein the ratio of the sedum aizoon extract solution to the macroporous resin is 10 mL: 1g of a compound;
(5) and (4) carrying out reduced pressure concentration on the supernatant obtained in the step (4) by using a rotary evaporator, recovering the solvent, heating to 45-50 ℃, obtaining a concentrated solution, namely the herba sedi aizoon total flavone, and freeze-drying for later use.
4. A preparation method of fermented milk beverage rich in GABA and anthocyanin is characterized by comprising the following steps:
(1) preparation method of bean sprouts rich in GABA and anthocyanin
Soaking seeds of the purple Chinese violet in a sodium hypochlorite solution of 12-18wt% for 3-5 hours, removing suspended particles, washing with pure water until the pH value is neutral, soaking in a mixed solution of 2.7-10.8g/L glucose and 0.5-1.5 g/L total flavonoids of the sedum aizoon at 28-32 ℃ for 22-26 hours, draining, placing on a gauze bed of a seed machine, continuously soaking with the mixed solution for germination under the conditions of 12-hour illumination and 12-hour dark treatment, replacing the soaking solution every day, taking out the roots after 7 days, cleaning the sprouts, draining, placing in a freeze dryer for 22-26 hours, and crushing to obtain sprout powder rich in GABA and anthocyanin;
(2) preparation of sugar-containing milk beverage
Soaking wheat grains in 28-32 deg.C water for 22-26 hr, germinating for 4 days, taking out, and cutting; soaking the corn particles in water for 3-5 hours, steaming at high temperature, cooling to 40-50 ℃, and taking out the corn particles; then mixing and fermenting the wheat crumbs and the corn crumbs according to the volume ratio of 1:10 for 5-6 hours, squeezing, concentrating to 1/5 of the original volume, and fixing the volume to 10 times of the volume of the concentrated solution by using pure milk to obtain the sugar-containing milk beverage;
(3) preparation of Lactobacillus plantarum suspensions
Inoculating lactobacillus plantarum into an MRS culture medium, and performing shake cultivation for 22-26 hours at 36-38 ℃ to obtain lactobacillus plantarum suspension;
(4) and (3) mixing the bud vegetable powder rich in GABA and anthocyanin prepared in the step (1) and the sugar-containing milk beverage prepared in the step (2) according to the equal volume, boiling, sealing, carrying out ultrasonic treatment at the temperature of 18-22 ℃ for 1-3 hours, filtering, adding the filtrate into the lactobacillus plantarum suspension prepared in the step (3), canning, preserving the heat at the temperature of 40-44 ℃ for 3-5 hours, and carrying out after-ripening at the temperature of 0-4 ℃ for 14-18 hours to obtain the fermented milk beverage rich in GABA and anthocyanin.
5. The method for preparing fermented milk beverage rich in GABA and anthocyanins according to claim 4, wherein the method comprises the following steps: the soak solution is a mixed solution containing 5.4g/L glucose and 1.0g/L herba Sedi total flavonoids.
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