CN101265487A - Preparation method for enriching gamma-aminobutyric acid by using fixed rice bran glutamic acid decarboxylase - Google Patents
Preparation method for enriching gamma-aminobutyric acid by using fixed rice bran glutamic acid decarboxylase Download PDFInfo
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- CN101265487A CN101265487A CNA2008100251362A CN200810025136A CN101265487A CN 101265487 A CN101265487 A CN 101265487A CN A2008100251362 A CNA2008100251362 A CN A2008100251362A CN 200810025136 A CN200810025136 A CN 200810025136A CN 101265487 A CN101265487 A CN 101265487A
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Abstract
The invention relates to a preparation method of enriching GABA by immobilized rice bran glutamate decarboxylase, and belongs to the technology field of the deep processing of the rice bran. In the method, the rice bran which is byproducts obtained after paddies are processed is used as raw material; the rice bran is mixed with sodium alginate solution with certain concentration and evenly stirred after being crushed and screened, and then is put in a balling device; CaCl2 solution is dropwise dropped into the mixed solution to act spherulization; ball bodies are sclerotized; the redundant CaCl2 and the rice bran which is not immobilized are washed, so that the immobilized rice bran glutamate decarboxylase is obtained; the immobilized rice bran glutamate decarboxylase is mixed with PBS buffer solution, sodium glutamate is added in and used as zymolyte, the concentration of the sodium glutamate is 0.01 to 0.05mol/l, the mixed solution reacts in a table concentrator under the temperature of 45 DEG C to obtain GABA enriched solution, and products are obtained after being dried. The protein content and the sugar content thereof are only a tenth part of the products which are obtained by adopting a traditional method of enriching the GABA by directly utilizing the rice bran; the products obtained by the method can be used as functional food additives; the efficacies of lowering blood pressure, calming, aiding sleeping, protecting liver, etc. can be realized; the preparation method has scientific and reasonable design, and has great significance to realize the comprehensive utilization of rice bran resources and increase the added value thereof.
Description
Technical field
The preparation method of a kind of enriching gamma-aminobutyric acid by using fixed rice bran glutamic acid decarboxylase (GABA) belongs to rice bran intensive processing technical field.
Background technology
Paddy is one of food crop main on the earth, and China usually is celebrated with " rice kingdom ", and the paddy annual production accounts for about 35% of world's paddy ultimate production, occupies first place in the world.Rice bran is as the byproduct of paddy, and it is very abundant to originate, and as fine utilization, can realize very high added value.L-glutamic acid cocarboxylase activity is very high in the rice bran, can be used for catalyzer, and catalysis monosodium glutamate generation decarboxylic reaction generates GABA.And the rice bran resource overwhelming majority of China consumes with traditional feed form, very waste.
The various food of the now existing high GABA content of Japan are sold in market, for example sprout brown rice, GABA tea, GABA skin care product, healthcare products etc., and market is fine, and China does not also have this type of commodity listing, suddenly treats that China production firm replenishes domestic blank.
Summary of the invention
The preparation method who the purpose of this invention is to provide fixed rice bran L-Glutamic decarboxylase enrichment GABA, utilize healthy nontoxic sodium alginate fixed rice bran L-Glutamic decarboxylase, conventional direct protein and the sugared content with rice bran enrichment GABA products obtained therefrom of products obtained therefrom declines to a great extent, only be the latter's 1/10, realize the enrichment of GABA and isolatingly carry out synchronously, saved the separation costs of GABA.And because the product foreign matter content greatly reduces, application wider.
Technical scheme of the present invention: a kind of preparation method of enriching gamma-aminobutyric acid by using fixed rice bran glutamic acid decarboxylase, earlier rice bran is pulverized, sieved, mixes with certain density sodium alginate soln and stir, the system of packing into ball equipment also dropwise splashes into CaCl
2Balling-up in the solution, the sclerosis spheroid cleans unnecessary CaCl
2The rice bran that is not immobilized gets the fixed rice bran L-Glutamic decarboxylase; This enzyme is mixed with PBS buffered soln, and the Sodium Glutamate that adds 0.05mol/L is as substrate, 45 ℃ of shaking tables react the γ-An Jidingsuan pregnant solution, oven dry promptly gets product;
(1) rice bran is pulverized, is sieved: earlier rice bran is pulverized, required 60 eye mesh screens, block the eyelet of system ball equipment to prevent larger particles.The raw material rice bran is wanted prepared fresh, prevents that the L-Glutamic decarboxylase enzyme activity of outmoded rice bran is too low.
(2) the back rice bran that sieves mixes with sodium alginate soln: earlier the preparation mass concentration is 2% sodium alginate soln, and sodium alginate dissolving back fully adds the rice bran that sieves, and makes the rice bran mass concentration of sieving control 6%, stirs, and gets mixed solution.
Because the sodium alginate solvability is relatively poor, can be lower than under 45 ℃ the condition heated and stirred to promote dissolving at solution temperature, after finishing, dissolving should use immediately or as early as possible, because of easily losing original viscosity after the placement of sodium alginate soln long period, until not wrapping rice bran.Rice bran and sodium alginate soln fully mixing are made ball again, mix abundant more, the parcel effect good more.
(3) system ball: the preparation mass concentration is the CaCl of 1%-1.5%
2Solution keeps solution stirring to flow; To go up the mixed solution of step (2) and put into system ball equipment, keep the speed that oozes of 1/s, splash into CaCl at distance liquid level 10cm place
2In the solution, balling-up immediately.
Keep CaCl
2The solution stirring velocity of flow is suitable, with the spheroid that falls into solution not the phase adhesion be advisable, the too fast spheroid that easily makes breaks; The speed that oozes of system ball equipment keeps 1/s, and is too fast or cross slow institute and become spheroid all not justify.
Drop keeps splashing into CaCl apart from liquid level 10cm place
2In the solution, balling-up immediately, too high or too low spheroid form and the immobilization effect of also influencing.
(4) sclerosis: keep CaCl
2Flow of solution so that spheroid wherein suspends, does not need too fastly or slow excessively, and CaCl is removed in the 1h hypsokinesis
2Solution, the time, the long enzyme that can reduce was lived, and CaCl is removed in the 1h hypsokinesis
2Solution.Strict control CaCl
2Strength of solution, CaCl
2The enzyme that has a strong impact on the fixed rice bran L-Glutamic decarboxylase is lived, and spheroid easily breaks during lower concentration, and Physical Mechanical is good inadequately, and high density can be lived by the strongly inhibited enzyme.
(5) clean: to equal CaCl
2The deionized water of liquor capacity is replaced CaCl
2Solution keeps stirring, and cleans spheroid, changes water altogether three times, stirs 20min at every turn; The purpose of cleaning is to remove not immobilized rice bran, and described scavenging period and number of times are enough, do not need additionally to add again.
Take during cleaning and carve the suspension of maintenance spheroid, could clean unnecessary CaCl
2, in case inhibitory enzyme is lived.
(6) rinse: deionized water is changed to PBS buffered soln, keeps stirring, the rinse spheroid changes PBS buffered soln altogether twice, stirs 20min at every turn, gets the immobilization L-Glutamic decarboxylase; The rinse operation promptly is the continuation of cleaning, and is again to prepare for reaction.This operation needs to keep spheroid to suspend constantly.
(7) reaction: with the immobilization L-Glutamic decarboxylase: PBS buffered soln mass ratio is controlled to be 3: 4 ratio in insulating container, the concentration of pressing 0.01-0.05mol/L adds Sodium Glutamate, keeps stirring reaction, 45 ℃ of temperature, react after 10-13h hour, get reaction solution;
The concentration of Sodium Glutamate can be regulated, the concentration of 0.01-0.05mol/L all can, the Sodium Glutamate of high density can strengthen enzyme lives, but the Sodium Glutamate add-on is big; The Sodium Glutamate enzyme of lower concentration is lived in increasing and is lacked, but the Sodium Glutamate add-on is few.Stirring velocity suspends with all spheroids and is advisable, and too fast or mistake is unfavorable for all that slowly Sodium Glutamate passes through the alginate calcium film, thereby reduces speed of reaction.
Reaction times can be regulated in 10-13h, and the time, too short GABA output was too low, and the GABA transaminase in the overlong time rice bran can cause that the GABA of generation decomposes.
(8) oven dry: remove the moisture in the reaction solution, promptly get the enriching gamma-aminobutyric product.Temperature is unsuitable too high in case the amino acid decomposition.
The preparation method of described enriching gamma-aminobutyric acid by using fixed rice bran glutamic acid decarboxylase, the raw material rice bran is the first fresh road rice bran; PBS buffered soln be formulated as 0.05mol/L KH
2PO
4And 0.05mol/LNa
2HPO
4Mixing solutions, the pH value is 5.6.
Beneficial effect of the present invention: that adopts the inventive method preparation is rich in the γ-An Jidingsuan product, under identical alpha-aminobutyric acid content condition, its protein content and sugared content only are traditional 1/10 of rice bran enriching gamma-aminobutyric method products obtained therefrom that directly utilizes, and the scope of application is more wide.Because foreign matter content is few, can be widely used in the production of health or functional food additives, healthcare products, medicine, makeup etc., the inventive method design science is reasonable, to the comprehensive utilization that realizes the rice bran resource with improve its added value and have great importance.
Description of drawings
Preparation technology's schematic flow sheet of Fig. 1 fixed rice bran L-Glutamic decarboxylase enrichment GABA.
Embodiment
Embodiment 1
Take by weighing the 20g sodium alginate and be dissolved in the 1L deionized water, the even sodium alginate soln that gets of stirring and dissolving, solution weight is as the part of immobilized enzyme gross weight; The raw material rice bran was through pulverizing 60 mesh sieves, took by weighing the 60g sieve back rice bran sodium alginate soln mixing and stirring good with dissolving, and the system of packing into ball device splashes into 1.5% CaCl apart from liquid level 10cm place with the speed of 1/s
2(contain CaCl in the solution
2The vessel whipping appts of solution, stirring velocity do not sink to the bottom with the spheroid in the solution and are advisable), the system ball.Behind system chou bundle, make particle in CaCl
2Stirring suspension 1h sclerosis in the solution.Remove CaCl behind the 1h
2Solution adds and CaCl
2The deionized water of solution equivalent is regulated and is continued to stir 20min, renews bright deionized water behind the 20min and stirs again, so uses deionized water rotating wash bowl body three times, and each 20min uses PBS buffered soln rinse spheroid twice again instead, each 20min; Remove the about 1.13kg of being fixed of PBS buffered soln enzyme.Immobilized enzyme is added 1.5L (about 1.5kg) PBS buffered soln, add 13.1g (0.07mol) Sodium Glutamate, be incubated 45 ℃ of stirring reaction 12h, remove immobilized enzyme behind the 12h, dry reaction liquid must be rich in the GABA product.
Embodiment 2
Take by weighing the 10g sodium alginate and be dissolved in the 0.5L deionized water, the even sodium alginate soln that gets of stirring and dissolving, solution weight is as the part of immobilized enzyme gross weight; The raw material rice bran took by weighing the 30g sodium alginate soln mixing and stirring good with dissolving through pulverizing 60 mesh sieves, and the system of packing into ball device splashes into 1% CaCl apart from liquid level 10cm place with the speed of 1/s
2Solution (is contained CaCl
2The vessel whipping appts of solution, stirring velocity do not sink to the bottom with the spheroid in the solution and are advisable) in, the system ball.Behind system chou bundle, make particle in CaCl
2Stirring suspension 1h sclerosis in the solution.Remove CaCl behind the 1h
2Solution adds and CaCl
2The deionized water of solution equivalent is regulated and is continued to stir 20min, renews bright deionized water behind the 20min and stirs again, so uses deionized water rotating wash bowl body three times, and each 20min uses PBS buffered soln rinse spheroid twice instead, each 20min; Remove the about 0.56kg of being fixed of PBS buffered soln enzyme.Immobilized enzyme is added 0.75L (about 0.75kg) PBS buffered soln, add 6.55g (0.035mol) Sodium Glutamate, be incubated 45 ℃ of stirring reaction 12h, remove immobilized enzyme behind the 12h, dry reaction liquid must be rich in the GABA product.
Claims (2)
1, a kind of preparation method of enriching gamma-aminobutyric acid by using fixed rice bran glutamic acid decarboxylase is characterized in that earlier rice bran being pulverized, being sieved, and mixes with certain density sodium alginate soln and stirs, and the system of packing into ball equipment also dropwise splashes into CaCl
2Balling-up in the solution, the sclerosis spheroid cleans unnecessary CaCl
2The rice bran that is not immobilized gets the fixed rice bran L-Glutamic decarboxylase; This enzyme is mixed with PBS buffered soln, and the Sodium Glutamate that adds 0.01-0.05mol/L is as substrate, 45 ℃ of shaking tables react the γ-An Jidingsuan pregnant solution, oven dry promptly gets product;
(1) rice bran is pulverized, is sieved: earlier rice bran is pulverized, required 60 eye mesh screens;
(2) the back rice bran that sieves mixes with sodium alginate soln: earlier the preparation mass concentration is 2% sodium alginate soln, and sodium alginate dissolving back fully adds the rice bran that sieves, and makes the rice bran mass concentration of sieving control 6%, stirs, and gets mixed solution;
(3) system ball: the preparation mass concentration is 1% CaCl
2Solution keeps solution stirring to flow; To go up the mixed solution of step (2) and put into system ball equipment, keep the speed that oozes of 1/s, splash into CaCl at distance liquid level 10cm place
2In the solution, balling-up immediately;
(4) sclerosis: keep CaCl
2Flow of solution, so that spheroid wherein suspends, CaCl is removed in the 1h hypsokinesis
2Solution;
(5) clean: to equal CaCl
2The deionized water of liquor capacity is replaced CaCl
2Solution keeps stirring, and cleans spheroid, changes water altogether three times, stirs 20min at every turn;
(6) rinse: deionized water is changed to PBS buffered soln, keeps stirring, the rinse spheroid changes PBS buffered soln altogether twice, stirs 20min at every turn, gets the immobilization L-Glutamic decarboxylase;
(7) reaction: with the immobilization L-Glutamic decarboxylase: PBS buffered soln mass ratio is controlled to be 3: 4 ratio in insulating container, and the concentration of press 0.01-0.05mol/L adds Sodium Glutamate, keeps stirring reaction, and 45 ℃ of temperature were reacted after 12 hours, must reaction solution;
(8) oven dry: remove the moisture in the reaction solution, promptly get the enriching gamma-aminobutyric product.
2,, it is characterized in that the raw material rice bran is the first fresh road rice bran according to the preparation method of the described enriching gamma-aminobutyric acid by using fixed rice bran glutamic acid decarboxylase of claim 1; PBS buffered soln be formulated as 0.05mol/LKH
2PO
4With 0.05mol/L Na
2HPO
4Mixing solutions, the pH value is 5.6.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676598A (en) * | 2012-05-30 | 2012-09-19 | 广东乐尔康生物科技股份有限公司 | Method for catalytically synthesizing gamma-aminobutyric acid by using sodium glutamate and immobilized bio-enzyme |
| CN102690846A (en) * | 2012-05-30 | 2012-09-26 | 广东乐尔康生物科技股份有限公司 | Method for catalytically synthesizing gamma-aminobutyric acid from glutamate biological solid-phase enzyme |
| CN103181523A (en) * | 2013-03-25 | 2013-07-03 | 南京财经大学 | Method for producing food material with high GABA function with rice endogenous enzyme activation method |
| CN103518942A (en) * | 2013-10-21 | 2014-01-22 | 江苏丹绿食品股份有限公司 | Production technology of rice bran protein powder |
| CN104872768A (en) * | 2015-05-27 | 2015-09-02 | 成都易创思生物科技有限公司 | Preparation process of GABA food beverage |
| CN104920810A (en) * | 2015-05-08 | 2015-09-23 | 李香子 | Preparation method of feed additive rich in gamma-aminobutyric acid |
| WO2021040085A1 (en) * | 2019-08-28 | 2021-03-04 | 주식회사 라파젠 | Composition for alleviating and treating narcolepsy, containing fermented rice bran powder as active ingredient, and preparation method therefor |
-
2008
- 2008-04-24 CN CNA2008100251362A patent/CN101265487A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102676598A (en) * | 2012-05-30 | 2012-09-19 | 广东乐尔康生物科技股份有限公司 | Method for catalytically synthesizing gamma-aminobutyric acid by using sodium glutamate and immobilized bio-enzyme |
| CN102690846A (en) * | 2012-05-30 | 2012-09-26 | 广东乐尔康生物科技股份有限公司 | Method for catalytically synthesizing gamma-aminobutyric acid from glutamate biological solid-phase enzyme |
| CN103181523A (en) * | 2013-03-25 | 2013-07-03 | 南京财经大学 | Method for producing food material with high GABA function with rice endogenous enzyme activation method |
| CN103181523B (en) * | 2013-03-25 | 2015-01-28 | 南京财经大学 | Method for producing food material with high GABA function with rice endogenous enzyme activation method |
| CN103518942A (en) * | 2013-10-21 | 2014-01-22 | 江苏丹绿食品股份有限公司 | Production technology of rice bran protein powder |
| CN104920810A (en) * | 2015-05-08 | 2015-09-23 | 李香子 | Preparation method of feed additive rich in gamma-aminobutyric acid |
| CN104872768A (en) * | 2015-05-27 | 2015-09-02 | 成都易创思生物科技有限公司 | Preparation process of GABA food beverage |
| WO2021040085A1 (en) * | 2019-08-28 | 2021-03-04 | 주식회사 라파젠 | Composition for alleviating and treating narcolepsy, containing fermented rice bran powder as active ingredient, and preparation method therefor |
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Open date: 20080917 |