CN108220351A - A kind of method that biological enzyme prepares L-arginine-α-ketoglutaric acid - Google Patents
A kind of method that biological enzyme prepares L-arginine-α-ketoglutaric acid Download PDFInfo
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Abstract
The invention discloses a kind of methods that biological enzyme prepares L arginine alpha Ketoglutarates, generate alpha Ketoglutarate using glucose oxidation enzymatic glutamic acid, add in arginine, prepare L arginine alpha Ketoglutarates（2:1）, and pass through ceramic membrane degerming, the removal of impurities of ultrafiltration membrane removing protein, activated carbon decolorizing, vacuum concentration, spray drying extract refined, this method is simple for process, and the smart ketone of production is high-quality, the basic zero-emission for realizing waste water.
Description
Technical field
The present invention relates to a kind of methods for producing smart ketone, and in particular to a kind of biological enzyme prepares L-arginine-α -one penta
The method of diacid.
Background technology
Smart ketone（L-arginine-α-ketoglutaric acid）Not only there is the physiological function of arginine and α-ketoglutaric acid, Er Qieke
To be applied to clinic as functional nutrient hardening agent and liver-protecting medicine, it is mainly used in enhancing physique, promotes the quick of muscle
Increase and wound is replied, promote absorption and metabolism of the liver cell to nutrition, enhance energy and muscle power, physiological Mechanism is mainly essence
Ketone can improve internal NO levels in a short time, enhance protein assimilation.At present, smart ketone as functional food in international city
Sales volume on field increasingly increases, and is had attracted much attention together with bird ketone in medicine, nutrition, Sports Field.Two kinds of smart ketone are main former
Material is arginine and α-ketoglutaric acid, the wherein big mature production technology of arginine, so be readily available, but α-ketoglutaric acid
It is more difficult to get.The preparation method of present α-ketoglutaric acid mainly has fermentation method and chemical synthesis, fermentation method production α -one penta 2
Generation pyruvic acid, fumaric acid, the by-products such as malic acid are more during acid, and fermentation period is long, so being difficult to obtain purity high
Product.Present α-ketoglutaric acid is mainly produced using chemical method, but using a large amount of extremely toxic substances in producing, production pollution is big,
It is dangerous height, complex process, of high cost.
Invention content
To make up the deficiencies in the prior art, the present invention provides a kind of biological enzyme and prepares L-arginine-α-ketoglutaric acid（2:
1）Method, this method be directly added into conversion fluid arginine synthesis L-arginine-α-ketoglutaric acid（2:1）, simple for process,
At low cost, L-arginine-α-ketoglutaric acid impurity of production is few, pollution is few, in current environmental protection pressure situation increasingly increased greatly
Under, it is highly beneficial to mass producing.
The present invention is achieved through the following technical solutions：
A kind of method that biological enzyme prepares L-arginine-α-ketoglutaric acid, is characterized in that：Using dglutamic oxidase
Glutamic acid generation α-ketoglutaric acid is catalyzed, arginine is added in, prepares L-arginine-α-ketoglutaric acid（2:1）, the letter of this method technique
Single, high-quality, pollution is small.
The method that a kind of biological enzyme of the present invention prepares L-arginine-α-ketoglutaric acid, includes the following steps：
（1）It is prepared by reaction solution：Glutamic acid is added in into pure water and arginine prepares reaction solution, adjusting reacting liquid pH value is 6.6-7；
（2）Enzyme reaction：Dglutamic oxidase is added in into reaction solution and catalase is reacted, reaction temperature 38-40
DEG C, reaction time 8-10h, pH value in reaction control is controlled in 6.5-7.0, dissolved oxygen in 5-30%；
（3）Ceramic membrane filter degerming：Reaction solution crosses ceramic membrane filter degerming, trapped fluid 2-4 times of volume of retention after reaction
Pure water dialyse；
（4）Ultrafiltration membrance filter removing protein cleans：The clear liquid that ceramic membrane filter obtains crosses ultrafiltration membrane, and ultrafiltration membrane pore diameter mol amount is
3000, trapped fluid is dialysed with the pure water of 2-4 times of volume of retention；
（5）Activated carbon decolorizing：The clear liquid that ultrafiltration membrance filter obtains is decolourized with activated carbon, 60-65 DEG C of bleaching temperature, during decoloration
Between 20-30min, add in activated carbon amount be 3-10g/L.
（6）It is concentrated under reduced pressure：Product after decoloration concentrates under-0.08-- 0.1mpa, is concentrated into L-arginine-α -one penta
A concentration of 40%-60% of diacid；
（7）Spray drying：It is spray-dried to obtain L-arginine-α-ketoglutaric acid product.
Preferably, step（1）Reaction solution Glutamic Acid a concentration of 100g/l, arginine concentrations 100g/l.
Preferably, step（2）The terminal of middle enzyme reaction is by measuring whether glutamic acid has residue to judge, paddy ammonia
It reacts and terminates after sour reaction completely.
Further, step（2）Middle reaction solution Glutamic Acid oxidizing ferment content is 3-5U/ml, and catalase content is
200-400U/ml。
Preferably, step（3）Bacterium mud drying after middle ceramic membrane filter is as feed addictive or feed egg
White raw material.
Preferably, step（4）Trapped fluid set after middle dialysis uses next group Synthesis liquid and crosses ceramic membrane together.
Preferably, step（5）The light transmittance T of solution after middle decoloration420nm≥95%。
Preferably, step（6）Middle thickening temperature is 65 DEG C.
Preferably, step（1）、（3）、（4）In pure water conductivity≤10.
The beneficial effects of the invention are as follows：
（1）α-ketoglutaric acid is to be made by glutamic acid by enzyme law catalysis, effectively reduces cost of material.
（2）There is no the addition of other impurities and inorganic salts in L-arginine-α-ketoglutaric acid building-up process, it is effective to improve
The product quality of L-arginine-α-ketoglutaric acid；L-arginine-α-ketoglutaric acid content of this method production reaches 99.0%
More than, meet every L-arginine-α-ketoglutaric acid quality standard.
（3）Ceramic membrane interception liquid is used for Fodder making albumen, and ultrafiltration membrane trapped fluid recycled, these all reduce waste water
Discharge.
Specific embodiment
The present invention will be further described in detail with reference to the specific embodiments, to help those skilled in the art
Inventive concept, technical solution to the present invention have more complete, accurate and deep understanding, and protection scope of the present invention is included but not
It is limited to following embodiment, any details to technical scheme of the present invention under the premise of without departing from spirit and scope
It is each fallen in protection scope of the present invention with the modification that form is made.
Embodiment 1
A kind of method that biological enzyme prepares L-arginine-α-ketoglutaric acid, includes the following steps：
（1）The preparation of reaction solution
Pure water 25m3 is added in into 50m3 reaction kettles, adds in 3000kg glutamic acid, 3650kg arginine adjusting pH value of solution to 6.9,
And it is settled to 30m3 with pure water.
（2）Enzyme reaction
The dglutamic oxidase of 3.5U/mL is added in into reaction solution, the catalase of 200U/mL starts to react, dissolved oxygen control
10%, temperature controls 39 ± 0.5 DEG C, and pH controls 6.9, reaction is detected with biosensor without glutamic acid after 10 hours, is added
3452kg arginine obtains L-arginine-α-ketoglutaric acid（2:1）Reaction solution.
（3）Ceramic membrane filter degerming
Reaction solution first crosses ceramic membrane after reaction, and trapped fluid is dialysed with the pure water of 4m3；Trapped fluid drying after dialysis is used
Forage protein is done, waste water is reduced and generates.
（4）Ultrafiltration membrance filter removing protein cleans
The clear liquid that ceramic membrane obtains crosses ultrafiltration membrane, and ultrafiltration membrane molecular weight is 3000 molecular weight, and trapped fluid is carried out with the pure water of 2m3
Analysis；Trapped fluid set after dialysis uses next group Synthesis liquid and crosses ceramic membrane together, cost-effective, and waste water is avoided to generate.
（5）Activated carbon decolorizing
65 DEG C of bleaching temperature, the amount for adding in activated carbon are 150kg, bleaching time 30min.The light transmission T of solution after decoloration420nm=
98.5%。
（6）It is concentrated under reduced pressure
65 DEG C of thickening temperature, pressure -0.1mpa are concentrated into 16m3.
（7）Spray drying
L-arginine-α-ketoglutaric acid is obtained after spray drying（2:1）Qualified products 10328kg, content 99.5%, yield are
95.4%。
Embodiment 2
A kind of method that biological enzyme prepares L-arginine-α-ketoglutaric acid, includes the following steps：
（1）The preparation of reaction solution
Pure water 25m3 is added in into 50m3 reaction kettles, then adds in 3000kg glutamic acid, 3640kg arginine, adjust pH value of solution to
6.9, and it is settled to 30m3 with pure water.
（2）Enzyme reaction
Dglutamic oxidase, the 250U/mL catalases that 5U/mL is added in into reaction solution start to react, dissolved oxygen control 20%,
Temperature controls 39.5 ± 0.5 DEG C, controls pH 6.9, and reaction is detected with biosensor without glutamic acid after 8 hours, adds arginine
3462kg obtains L-arginine-α-ketoglutaric acid（2:1）Reaction solution.
（3）Ceramic membrane filter degerming
Reaction solution first crosses ceramic membrane after reaction, and trapped fluid is dialysed with the pure water of 6m3；Trapped fluid drying after dialysis is used
Do forage protein.
（4）Ultrafiltration membrance filter removing protein cleans
The clear liquid that ceramic membrane obtains crosses ultrafiltration membrane, and ultrafiltration membrane molecular weight is 3000 molecular weight, and trapped fluid is carried out with the pure water of 3m3
Analysis；Trapped fluid set after dialysis uses next group Synthesis liquid and crosses ceramic membrane together.
（5）Activated carbon decolorizing
60 DEG C of bleaching temperature adds in the amount of activated carbon as 180kg, bleaching time 25min, the light transmission T of the solution after decoloration420nm=
98.1%。
（6）It is concentrated under reduced pressure
65 DEG C of thickening temperature, pressure -0.1mpa.It is concentrated into 15m3.
（7）Spray drying
L-arginine-α-ketoglutaric acid is obtained after spray drying（2:1）Qualified products 10360kg, content 99.4%, yield are
95.7%。
Embodiment 3
A kind of method that biological enzyme prepares L-arginine-α-ketoglutaric acid, includes the following steps：
（1）The preparation of reaction solution
Pure water 25m3 is added in into 50m3 reaction kettles, then adds in 3000kg glutamic acid, 3658kg arginine, adjust pH value of solution to
7.0, and it is settled to 30m3 with pure water.
（2）Enzyme reaction
Dglutamic oxidase, the 400U/mL catalases that 5U/mL is added in into reaction solution start to react, dissolved oxygen control 25%,
Temperature controls 38.5 ± 0.5 DEG C, and stream plus arginine control pH 7.0, reaction are detected with biosensor without paddy ammonia after 10 hours
Acid adds arginine 3444kg, obtains L-arginine-α-ketoglutaric acid（2:1）Reaction solution.
（3）Ceramic membrane filter degerming
Reaction solution first crosses ceramic membrane after reaction, and trapped fluid is dialysed with the pure water of 4m3；Trapped fluid drying after dialysis is used
Do forage protein.
（4）Ultrafiltration membrance filter removing protein cleans
The clear liquid that ceramic membrane obtains crosses ultrafiltration membrane, and ultrafiltration membrane molecular weight is 3000 molecular weight, and trapped fluid is carried out with the pure water of 2m3
Analysis；Trapped fluid set after dialysis uses next group Synthesis liquid and crosses ceramic membrane together.
（5）Activated carbon decolorizing
60 DEG C of bleaching temperature adds in the amount of activated carbon as 240kg, bleaching time 30min, the light transmission T of the solution after decoloration420nm=
97.1%。
（6）It is concentrated under reduced pressure
65 DEG C of thickening temperature, pressure -0.1mpa.It is concentrated into 18m3.
（7）Spray drying
L-arginine-α-ketoglutaric acid is obtained after spray drying（2:1）Qualified products 10253kg, content 99.5%, yield are
94.7%。
Claims (10)
1. a kind of method that biological enzyme prepares L-arginine-α-ketoglutaric acid, it is characterised in that：It is urged using dglutamic oxidase
Change glutamic acid generation α-ketoglutaric acid, add in arginine, prepare L-arginine-α-ketoglutaric acid.
2. the method that a kind of biological enzyme according to claim 1 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Include the following steps：
（1）It is prepared by reaction solution：Glutamic acid is added in into pure water and arginine prepares reaction solution, adjusting reacting liquid pH value is 6.6-7；
（2）Enzyme reaction：Dglutamic oxidase is added in into reaction solution and catalase is reacted, reaction temperature 38-40
DEG C, reaction time 8-10h, pH value in reaction control is controlled in 6.5-7.0, dissolved oxygen in 5-30%；
（3）Ceramic membrane filter：Reaction solution crosses ceramic membrane filter degerming after reaction, and trapped fluid is pure with 2-4 times of volume of retention
Water is dialysed；
（4）Ultrafiltration membrance filter：The clear liquid that ceramic membrane filter obtains crosses ultrafiltration membrane, and ultrafiltration membrane pore diameter mol amount is 3000, trapped fluid
It is dialysed with the pure water of 2-4 times of volume of retention；
（5）Activated carbon decolorizing：The clear liquid that ultrafiltration membrance filter obtains is decolourized with activated carbon, 60-65 DEG C of bleaching temperature, during decoloration
Between 20-30min, add in activated carbon amount be 3-10g/L；
（6）It is concentrated under reduced pressure：Product after decoloration concentrates under-0.08-- 0.1mpa, is concentrated into L-arginine-α-ketoglutaric acid
A concentration of 40%-60%；
（7）Spray drying：It is spray-dried to obtain L-arginine-α-ketoglutaric acid product.
3. the method that a kind of biological enzyme according to claim 2 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Step（1）Reaction solution Glutamic Acid a concentration of 100g/l, arginine concentrations 100g/l.
4. the method that a kind of biological enzyme according to claim 2 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Step（2）The terminal of middle enzyme reaction reacts knot by measuring whether glutamic acid has residue to judge after glutamic acid reaction completely
Beam.
5. the method that a kind of biological enzyme according to claim 4 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Step（2）Middle reaction solution Glutamic Acid oxidizing ferment content is 3-5U/ml, and catalase content is 200-400U/ml.
6. the method that a kind of biological enzyme according to claim 2 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Step（3）Bacterium mud drying after middle ceramic membrane filter is as feed addictive or the raw material of forage protein.
7. the method that a kind of biological enzyme according to claim 2 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Step（4）Trapped fluid set after middle dialysis uses next group Synthesis liquid and crosses ceramic membrane together.
8. the method that a kind of biological enzyme according to claim 2 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Step（5）The light transmittance T of solution after middle decoloration420nm≥95%。
9. the method that a kind of biological enzyme according to claim 2 prepares L-arginine-α-ketoglutaric acid, feature exist
In：Step（6）Middle thickening temperature is 65 DEG C.
10. a kind of biological enzyme according to any one of claim 2-9 prepares the side of L-arginine-α-ketoglutaric acid
Method, it is characterised in that：Step（1）、（3）、（4）In pure water conductivity≤10.
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| CN109851491A (en) * | 2019-04-16 | 2019-06-07 | 同舟纵横（厦门）流体技术有限公司 | A kind of method and device of gulonate feed liquid decoloration removal of impurities |
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| CN102020593B (en) * | 2010-11-29 | 2012-01-18 | 广东环西生物科技股份有限公司 | Process for preparing L-arginine-alpha-ketoglutarate (AAKG) from fermentation liquor through direct crystallization |
| CN104109698A (en) * | 2013-04-17 | 2014-10-22 | 上海工业生物技术研发中心 | Enzymic method for producing [alpha]-ketoglutaric acid |
| CN106350547A (en) * | 2016-08-24 | 2017-01-25 | 天津科技大学 | Preparation method of L-arginine-alpha-ketoglutaric acid |
| CN106148434A (en) * | 2016-08-27 | 2016-11-23 | 山东民强生物科技股份有限公司 | A kind of microbial enzyme method produces the method for a ketoglutaric acid |
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| CN109851491A (en) * | 2019-04-16 | 2019-06-07 | 同舟纵横（厦门）流体技术有限公司 | A kind of method and device of gulonate feed liquid decoloration removal of impurities |
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Denomination of invention: Preparation of L-arginine by biological enzymatic method- a- Method for producing ketoglutarate Effective date of registration: 20220530 Granted publication date: 20210914 Pledgee: Shandong juancheng Rural Commercial Bank Co.,Ltd. Plaza sub branch Pledgor: SHANDONG YANGCHENG BIOTECH Co.,Ltd. Registration number: Y2022980006634 |