CN107648389B - Preparation method of banana aqueous extract containing gamma-aminobutyric acid - Google Patents
Preparation method of banana aqueous extract containing gamma-aminobutyric acid Download PDFInfo
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- CN107648389B CN107648389B CN201710801337.6A CN201710801337A CN107648389B CN 107648389 B CN107648389 B CN 107648389B CN 201710801337 A CN201710801337 A CN 201710801337A CN 107648389 B CN107648389 B CN 107648389B
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- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 title claims abstract description 86
- 235000018290 Musa x paradisiaca Nutrition 0.000 title claims abstract description 57
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 229960003692 gamma aminobutyric acid Drugs 0.000 title claims abstract description 43
- 239000006286 aqueous extract Substances 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 240000005561 Musa balbisiana Species 0.000 title 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims abstract description 75
- 239000004220 glutamic acid Substances 0.000 claims abstract description 64
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims abstract description 62
- 235000013922 glutamic acid Nutrition 0.000 claims abstract description 62
- 241000234295 Musa Species 0.000 claims abstract description 56
- 239000000203 mixture Substances 0.000 claims abstract description 54
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 37
- 238000006243 chemical reaction Methods 0.000 claims abstract description 34
- 238000003756 stirring Methods 0.000 claims abstract description 32
- 238000000034 method Methods 0.000 claims abstract description 23
- 238000001914 filtration Methods 0.000 claims abstract description 14
- 239000000284 extract Substances 0.000 claims abstract description 13
- 229930195712 glutamate Natural products 0.000 claims abstract description 13
- 230000000694 effects Effects 0.000 claims abstract description 11
- 239000000706 filtrate Substances 0.000 claims abstract description 11
- 102000008214 Glutamate decarboxylase Human genes 0.000 claims abstract description 10
- 108091022930 Glutamate decarboxylase Proteins 0.000 claims abstract description 10
- 238000001035 drying Methods 0.000 claims abstract description 10
- 230000001502 supplementing effect Effects 0.000 claims abstract description 9
- 238000012544 monitoring process Methods 0.000 claims abstract description 7
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims description 23
- 239000007787 solid Substances 0.000 claims description 20
- 239000006228 supernatant Substances 0.000 claims description 18
- 239000012528 membrane Substances 0.000 claims description 9
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 8
- 239000002244 precipitate Substances 0.000 claims description 6
- 239000011541 reaction mixture Substances 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 3
- 235000013399 edible fruits Nutrition 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 235000013919 monopotassium glutamate Nutrition 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 abstract description 8
- 108090000790 Enzymes Proteins 0.000 abstract description 8
- 239000007853 buffer solution Substances 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 5
- 229910017053 inorganic salt Inorganic materials 0.000 abstract description 3
- 238000004904 shortening Methods 0.000 abstract 1
- 229940049906 glutamate Drugs 0.000 description 9
- QKFJKGMPGYROCL-UHFFFAOYSA-N phenyl isothiocyanate Chemical compound S=C=NC1=CC=CC=C1 QKFJKGMPGYROCL-UHFFFAOYSA-N 0.000 description 8
- 235000013923 monosodium glutamate Nutrition 0.000 description 5
- 229940073490 sodium glutamate Drugs 0.000 description 5
- 229940117953 phenylisothiocyanate Drugs 0.000 description 4
- 230000035484 reaction time Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000012510 hollow fiber Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 208000019901 Anxiety disease Diseases 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- -1 after being crushed Substances 0.000 description 1
- 230000036506 anxiety Effects 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 208000029028 brain injury Diseases 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000012047 saturated solution Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/175—Amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Abstract
The invention discloses a preparation method of a banana water extract containing gamma-aminobutyric acid, which comprises the following steps: crushing banana pulp into homogenate, adding glutamate and glutamic acid, stirring and mixing to obtain a mixture with the pH value of 5.0-5.7, stirring and reacting at the temperature of 25-40 ℃, monitoring the change of the pH value of the mixture in the reaction process, supplementing the glutamic acid when the pH value is higher than 5.7 every time, continuously maintaining the pH value of the mixture between 5.0-5.7, reacting for 12-24 hours until the activity of glutamate decarboxylase disappears, adding water into the mixture after the reaction is finished, stirring and extracting, filtering, concentrating the filtrate, and drying at the temperature of 40-80 ℃ to obtain the banana aqueous extract containing gamma-aminobutyric acid. The invention does not need to carry out the step of extracting glutamic acid decarboxylase, namely, does not need to use the buffer solution of inorganic salt, thereby shortening the process steps, improving the process efficiency, improving the utilization rate of enzyme, and the used raw materials can be eaten, so that the prepared extract containing the gamma-aminobutyric acid is safer to eat.
Description
The technical field is as follows:
the invention relates to the technical field of biological enzyme synthesis, in particular to a preparation method of a banana water extract containing gamma-aminobutyric acid.
Background art:
glutamate decarboxylase in banana can catalyze glutamic acid to remove a carboxyl group and convert the glutamic acid into gamma-aminobutyric acid and carbon dioxide. Gamma-aminobutyric acid has the effects of reducing blood pressure, improving sleep, resisting anxiety, repairing brain injury, improving memory and the like, and is an important raw material for medicaments and health-care foods.
In the prior patent method (application No. 201310122628.4), banana is taken as a raw material, after being crushed, phosphate buffer salt, calcium chloride and the like are used for extracting glutamic acid decarboxylase in the banana, and then the obtained product reacts with glutamic acid and sodium glutamate to prepare the gamma-aminobutyric acid.
In view of the above, the present inventors propose the following.
The invention content is as follows:
the invention aims to overcome the defects of the prior art and provides a preparation method of a banana aqueous extract containing gamma-aminobutyric acid.
In order to solve the technical problems, the invention adopts the following technical scheme: the preparation method of the banana water extract containing gamma-aminobutyric acid comprises the following steps: crushing banana pulp into homogenate, adding glutamate and glutamic acid, stirring and mixing to obtain a mixture with the pH value of 5.0-5.7, stirring and reacting at the temperature of 25-40 ℃, monitoring the change of the pH value of the mixture in the reaction process, supplementing the glutamic acid when the pH value is higher than 5.7 every time, continuously maintaining the pH value of the mixture between 5.0-5.7, reacting for 12-24 hours until the activity of glutamate decarboxylase disappears, adding water into the mixture after the reaction is finished, stirring and extracting, filtering, concentrating the filtrate, and drying at the temperature of 40-80 ℃ to obtain the banana aqueous extract containing gamma-aminobutyric acid.
Further, in the technical scheme, the banana pulp is obtained by peeling off the peel of the domestic or imported mature banana fruits.
Furthermore, in the above technical solution, 15-25 g glutamate is added per 100 g banana pulp homogenate.
Further, in the above technical solution, the glutamate is a solid powder of glutamic acid-sodium salt or glutamic acid-potassium salt.
Further, in the above technical solution, the glutamic acid is solid powder, and the glutamic acid is added in an amount that the added solid powder of glutamic acid is not dissolved in the banana homogenate in addition to a small amount of the solid powder of glutamic acid.
Furthermore, in the above technical scheme, after the glutamic acid in the mixture is reacted, the pH value of the mixture is rapidly increased, and when the pH value is higher than 5.7, the glutamic acid is supplemented for multiple times, so long as the existence of solid glutamic acid in the banana homogenate is ensured, the pH value of the reaction mixture can be maintained between 5.0 and 5.7.
Further, in the above technical scheme, the temperature of the stirring reaction is 30-35 ℃.
Further, in the above technical scheme, the filtering comprises adding water, stirring, extracting, performing solid-liquid separation by centrifugation, and filtering the separated aqueous solution with an ultra-micro filtration membrane.
Further, in the above technical scheme, after the reaction is completed, adding water into the mixture at normal temperature, stirring for 30 minutes, centrifuging for 5 minutes at a speed of 3000 rpm, taking the supernatant, adding water into the precipitate, stirring for 15 minutes, centrifuging for 5 minutes at a speed of 3000 rpm, taking the supernatant, combining, filtering the supernatant with an ultramicro filter membrane of 0.001 micron to obtain a filtrate, concentrating the filtrate under reduced pressure to obtain a thick paste, and drying the thick paste under vacuum at 60-80 ℃ to obtain the banana water extract containing gamma-aminobutyric acid.
After adopting the technical scheme, compared with the prior art, the invention has the following beneficial effects: the preparation method of the banana aqueous extract containing gamma-aminobutyric acid does not need a glutamic acid decarboxylase leaching step, namely, does not need to use a buffer solution of inorganic salt, shortens the process steps, improves the process efficiency, improves the utilization rate of enzyme, and in addition, all the used raw materials can be eaten in the preparation process of the gamma-aminobutyric acid, so that the prepared extract containing the gamma-aminobutyric acid is safer to eat.
The specific implementation mode is as follows:
the present invention is further illustrated by the following specific examples.
The invention relates to a preparation method of a banana water extract containing gamma-aminobutyric acid, which comprises the following steps: crushing banana pulp into homogenate, adding glutamate and glutamic acid, stirring and mixing to obtain a mixture with the pH value of 5.0-5.7, stirring and reacting at the temperature of 25-40 ℃, monitoring the change of the pH value of the mixture in the reaction process, supplementing the glutamic acid when the pH value is higher than 5.7 every time, continuously maintaining the pH value of the mixture between 5.0-5.7, reacting for 12-24 hours until the activity of glutamate decarboxylase disappears, adding water into the mixture after the reaction is finished, stirring and extracting, filtering, concentrating the filtrate, and drying at the temperature of 40-80 ℃ to obtain the banana aqueous extract containing gamma-aminobutyric acid.
The banana pulp is obtained by peeling off the peel of domestic or imported mature banana fruits.
Glutamic acid is an organic acid which is difficult to dissolve in water, the solubility of glutamic acid in the mixture is low, solid glutamic acid added is mostly in a solid form in the mixture, and a small amount of glutamic acid is dissolved in the banana homogenate, so that the mixture becomes a glutamic salt-glutamic acid buffer system, and the pH value is maintained between 5.0 and 5.7. The amount of glutamic acid added does not affect the pH of the mixture as long as solid glutamic acid is present in the mixture, and the less the amount of glutamic acid added, the more frequent the addition.
Since glutamate is readily soluble in water, the glutamate added to the banana homogenate is totally dissolved in the aqueous phase of the mixture, with more glutamate added the higher the pH of the mixture. Adding 15-25 g glutamate per 100 g banana pulp homogenate, wherein the glutamate is solid powder of glutamic acid-sodium salt or glutamic acid-potassium salt. The glutamic acid is difficult to dissolve in water, only a small amount of added glutamic acid is dissolved in the water phase of the mixture to form a glutamic acid saturated solution, most of the glutamic acid is still in a solid state in the solid phase of the mixture, the solid glutamic acid can continuously supplement the consumption of converting the glutamic acid in the water phase of the mixture into the gamma-aminobutyric acid, and the stability of the concentration of the glutamic acid in the water phase of the mixture is maintained, so that the stable pH value is maintained, and a glutamate-glutamic acid buffer system is formed.
Glutamic acid is a solid powder, and the glutamic acid is added in such an amount that the added solid powder of glutamic acid has not been dissolved in addition to a small amount of the dissolved solid powder of glutamic acid in the banana homogenate.
When the glutamic acid powder in the mixture is completely consumed by reaction, the pH value of the mixture is rapidly increased, and the original pH value buffer system is restored by supplementing the glutamic acid powder. The amount of glutamic acid in the solid phase of the mixture does not affect the pH value of the mixture, but only affects the frequency of glutamic acid supplement.
After the glutamic acid in the mixture is reacted, the pH value of the mixture is quickly increased, when the pH value is higher than 5.7, the glutamic acid is supplemented for many times, and the pH value of the reaction mixture can be maintained between 5.0 and 5.7 as long as the solid glutamic acid exists in the banana homogenate. Specifically, the activity of enzyme is strong when the reaction of glutamic acid in the mixture is started, the reaction speed is high, the glutamic acid added for the first time can be 5-10% of the mass of the banana homogenate, the amount of the glutamic acid added for each time in the later period of the reaction is preferably less, and the glutamic acid can be added according to 5-1% of the mass of the banana homogenate.
The time of the reaction is related to the temperature of the reaction, the higher the temperature, the faster the activity of the enzyme decays. The reaction temperature is 25 ℃, the enzyme activity is almost lost after 24 hours of reaction, the enzyme activity is almost lost after 12 hours of reaction at 40 ℃, the optimal reaction temperature is 30-35 ℃, the glutamate decarboxylase activity is highest at 30-35 ℃, the effect is best, and the optimal reaction time is 12-24 hours.
The filtration comprises adding water, stirring, extracting, performing solid-liquid separation by centrifugation, and filtering the separated water solution with ultramicro filter membrane. More specifically, after the reaction is finished, adding water into the mixture at normal temperature, stirring for 30 minutes, centrifuging for 5 minutes at the speed of 3000 rpm, taking the supernatant, adding water into the precipitate, stirring for 15 minutes, centrifuging for 5 minutes at the speed of 3000 rpm, taking the supernatant, combining, passing the supernatant through an ultramicro filter membrane of 0.001 micron to obtain filtrate, concentrating under reduced pressure to obtain thick paste, and drying the thick paste in vacuum at the temperature of 60-80 ℃ to obtain the banana water extract containing gamma-aminobutyric acid.
Influence of different reaction temperatures and different reaction times on the amount of produced gamma-aminobutyric acid
Preparing mature banana pulp into banana pulp, taking 50 g of homogenate per part, putting the homogenate into a 100ml flask, respectively adding 10 g of sodium glutamate and 5 g of glutamic acid, stirring and reacting at 25 ℃, 27, 30, 33, 35, 37 and 40 ℃ respectively, and respectively reacting for 4 hours, and supplementing 2.5 g of glutamic acid each time for 8 hours. After 8, 12, 16, 20 and 24 hours, 1 g of the reaction mixture is put into a 20ml test tube, 10ml of water is added, the mixture is shaken and heated in boiling water for 15 minutes, then the mixture is transferred to a 500ml measuring flask, and the water is added to the scale mark. And (3) measuring the content of the gamma-aminobutyric acid by a phenyl isothiocyanate method, and calculating the amount (mg) of the gamma-aminobutyric acid generated in each gram of mixture at different reaction temperatures and different reaction times. The results are shown in Table 1:
table 1 content of gamma-aminobutyric acid (content of gamma-aminobutyric acid mg/g of fermentation product) produced in the mixture at different reaction temperatures for different periods of time.
It can be seen that the optimum reaction (fermentation) temperature is 30-35 deg.C, and the optimum reaction time is 12-24 hr at 30-35 deg.C.
The preparation method of the banana aqueous extract containing gamma-aminobutyric acid does not need a glutamic acid decarboxylase leaching step, namely, does not need to use a buffer solution of inorganic salt, shortens the process steps, improves the process efficiency, improves the utilization rate of enzyme, and in addition, all the used raw materials can be eaten in the preparation process of the gamma-aminobutyric acid, so that the prepared extract containing the gamma-aminobutyric acid is safer to eat.
Example 1:
crushing mature banana pulp into homogenate, taking 500 g of homogenate, placing the homogenate in a 1000ml flask, adding 100 g of sodium glutamate powder, stirring to dissolve, adding 50 g of glutamic acid powder, stirring, and reacting the mixture under stirring at the constant temperature of 33 ℃. Monitoring the change of the pH value in the reaction process, and supplementing a proper amount of glutamic acid powder when the pH value is higher than 5.7 so as to continuously maintain the pH value of the mixture between 5.0 and 5.7. After 20 hours of reaction, 1000ml of water was added, the mixture was stirred for 30 minutes, centrifuged at 3000 rpm for 5 minutes, and the supernatant was collected. The precipitate was stirred with 1000ml of water for 30 minutes, centrifuged at 3000 rpm for 5 minutes, and the supernatants were combined. Filtering the supernatant with a hollow fiber filter membrane with the aperture of 0.001 micron, concentrating the filtrate, and drying at 40-80 ℃ to obtain 205.4 g of the banana aqueous extract containing the gamma-aminobutyric acid. The content of gamma-aminobutyric acid in the extract is 31.8% by phenyl isothiocyanate method.
Example 2:
crushing mature banana pulp into homogenate, taking 500 g of homogenate, placing the homogenate in a 1000ml flask, adding 75 g of sodium glutamate powder, stirring to dissolve, adding 50 g of glutamic acid powder, and stirring. Stirring the mixture at a constant temperature of 30 ℃ for reaction, monitoring the change of pH value in the reaction process, and supplementing a proper amount of glutamic acid powder when the pH value is higher than 5.7 so as to continuously maintain the pH value of the mixture between 5.0 and 5.7. After 24 hours of reaction, 1000ml of water was added, the mixture was stirred for 30 minutes, centrifuged at 3000 rpm for 5 minutes, and the supernatant was collected. The precipitate was stirred with 1000ml of water for 30 minutes, centrifuged at 3000 rpm for 5 minutes, and the supernatants were combined. Filtering the supernatant with a hollow fiber filter membrane with the aperture of 0.001 micron, concentrating the filtrate, and drying at 40-80 ℃ to obtain 180.7 g of the banana aqueous extract containing gamma-aminobutyric acid. The content of gamma-aminobutyric acid in the extract is 34.1% by phenyl isothiocyanate method.
Example 3:
crushing mature banana pulp into homogenate, taking 500 g of homogenate, placing the homogenate in a 1000ml flask, adding 125 g of sodium glutamate powder, stirring to dissolve, adding 50 g of glutamic acid powder, and stirring. The mixture was stirred for reaction at a constant temperature of 35 ℃. Monitoring the change of the pH value in the reaction process, and supplementing a proper amount of glutamic acid powder when the pH value is higher than 5.7 so as to continuously maintain the pH value of the mixture between 5.0 and 5.7. After 12 hours of reaction, 1000ml of water was added, the mixture was stirred for 30 minutes, centrifuged at 3000 rpm for 5 minutes, and the supernatant was collected. The precipitate was stirred with 1000ml of water for 30 minutes, centrifuged at 3000 rpm for 5 minutes, and the supernatants were combined. Filtering the supernatant with a hollow fiber filter membrane with the aperture of 0.001 micron, concentrating the filtrate, and drying at 40-80 ℃ to obtain 234.7 g of the banana aqueous extract containing the gamma-aminobutyric acid. The content of gamma-aminobutyric acid in the extract is 25.2% by phenyl isothiocyanate method.
It should be understood that the above description is only exemplary of the present invention, and is not intended to limit the scope of the present invention, which is defined by the appended claims.
Claims (7)
1. A preparation method of a banana aqueous extract containing gamma-aminobutyric acid is characterized by comprising the following steps: which comprises the following steps: crushing banana pulp into homogenate, adding glutamate and glutamic acid, stirring and mixing to obtain a mixture with the pH value of 5.0-5.7, stirring and reacting at the temperature of 25-40 ℃, monitoring the change of the pH value of the mixture in the reaction process, supplementing the glutamic acid when the pH value is higher than 5.7 every time, continuously maintaining the pH value of the mixture between 5.0-5.7, reacting for 12-24 hours until the activity of glutamate decarboxylase disappears, adding water into the mixture after the reaction is finished, stirring and extracting, filtering, concentrating the filtrate, and drying at the temperature of 40-80 ℃ to obtain the banana aqueous extract containing gamma-aminobutyric acid;
adding glutamate 15-25 g per 100 g banana pulp homogenate;
the filtration comprises adding water, stirring, extracting, performing solid-liquid separation by centrifugation, and filtering the separated water solution with ultramicro filter membrane.
2. The method for preparing aqueous extract of banana containing gamma-aminobutyric acid as claimed in claim 1, wherein the method comprises the following steps: the banana pulp is obtained by peeling off the peel of domestic or imported mature banana fruits.
3. The method for preparing aqueous extract of banana containing gamma-aminobutyric acid as claimed in claim 1, wherein the method comprises the following steps: the glutamate is solid powder of glutamic acid-sodium salt or glutamic acid-potassium salt.
4. The method for preparing aqueous extract of banana containing gamma-aminobutyric acid as claimed in claim 1, wherein the method comprises the following steps: glutamic acid is a solid powder, and the glutamic acid is added in such an amount that the added solid powder of glutamic acid has not been dissolved in addition to a small amount of the dissolved solid powder of glutamic acid in the banana homogenate.
5. The method for preparing aqueous extract of banana containing gamma-aminobutyric acid as claimed in claim 1, wherein the method comprises the following steps: after the glutamic acid in the mixture is reacted, the pH value of the mixture is quickly increased, when the pH value is higher than 5.7, the glutamic acid is supplemented for many times, and the pH value of the reaction mixture can be maintained between 5.0 and 5.7 as long as the solid glutamic acid exists in the banana homogenate.
6. The method for preparing aqueous extract of banana containing gamma-aminobutyric acid as claimed in claim 1, wherein the method comprises the following steps: the temperature of the stirring reaction is 30-35 ℃.
7. The method for preparing aqueous extract of banana containing gamma-aminobutyric acid as claimed in claim 1, wherein the method comprises the following steps: after the reaction is finished, adding water into the mixture at normal temperature, stirring for 30 minutes, centrifuging at the speed of 3000 rpm for 5 minutes, taking supernatant, adding water into the precipitate, stirring for 15 minutes, centrifuging at the speed of 3000 rpm for 5 minutes, taking supernatant, combining, passing the supernatant through an ultramicro filter membrane of 0.001 micron to obtain filtrate, concentrating under reduced pressure to obtain thick paste, and drying the thick paste in vacuum at the temperature of 60-80 ℃ to obtain the banana water extract containing the gamma-aminobutyric acid.
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| JP2008245527A (en) * | 2007-03-29 | 2008-10-16 | Acera:Kk | Method for producing food material containing large amount of gamma-aminobutyric acid, method for producing gamma-aminobutyric acid enriched food, food material containing large amount of gamma-aminobutyric acid, gamma-aminobutyric acid enriched food, and food material for producing gamma-aminobutyric acid |
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