CN104560707B - The process of a kind of Enzyme catalyzed synthesis AA 2G and process system - Google Patents
The process of a kind of Enzyme catalyzed synthesis AA 2G and process system Download PDFInfo
- Publication number
- CN104560707B CN104560707B CN201510063451.4A CN201510063451A CN104560707B CN 104560707 B CN104560707 B CN 104560707B CN 201510063451 A CN201510063451 A CN 201510063451A CN 104560707 B CN104560707 B CN 104560707B
- Authority
- CN
- China
- Prior art keywords
- defecator
- resin column
- reactor
- drinking water
- centrifuged
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/02—Membranes; Filters
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- Molecular Biology (AREA)
- Clinical Laboratory Science (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention provides process system and the process of a kind of Enzyme catalyzed synthesis L ascorbic acid 2 glucoside, including reactor (1), enzyme basin (2), basin (3), centrifuge (4), the first defecator (5), the first resin column (6), the second resin column (7), the second defecator (8), falling film concentrating device (9), the 3rd defecator (10)；The process of the present invention, including enzyme reaction, the preparation of L ascorbic acid 2 glucoside crude product and crude product refining.The present invention effectively goes the removal of impurity, improves the purity of finished product；Simple to equipment requirements；Yield is high, and post processing is simple, low cost.
Description
Technical field
The present invention relates to a kind of process preparing AA 2G and process system, the process of a kind of Enzyme catalyzed synthesis AA 2G and process system.
Background technology
AA 2G is (also known as vitamin C glucoside, it is called for short AA-2G, L-VCG), the entitled 2-oxygen-α-D-glucopyranosyl ascorbic acid of chemistry, it it is ascorbic carbohydrate derivative, cause has the following advantages and becomes ascorbic best substitute: 1) good non-reduced activity, stable in aqueous；2) good thermostability and light resistance；3) produce vitamin C and glucose in intracellular hydrolysis, there is the biological activity equal with vitamin C.
AA 2G is one of 6 kinds of whitening additives of Department of Health's announcement accreditation.Melanin that it can be expelled directly out having been formed in epidermal area (will established melanin add speed in reversed direction be reduced to initial form), whitening effect becomes apparent from than other additives and quickly, therefore suffers from being widely applied in multiple high-end skin-lightening cosmetic.Existing preparing in AA 2G technique, impurity difficulty is removed, and AA 2G purity is the highest, and yield is low.
Summary of the invention
In order to solve the problems referred to above, first purpose of the present invention is to provide a kind of process system improving AA 2G purity.
Second purpose of invention is to provide a kind of process improving AA 2G purity.
The cyclodextrin glycosyl transferases (abbreviation CGT enzyme) that the present invention provides uses the prepared offer of preparation method of Patent No. 201410236224.2 cyclodextrin glycosyl transferase.
Technical scheme:
The process system of a kind of Enzyme catalyzed synthesis AA 2G, including reactor (1), enzyme basin (2), basin (3), centrifuge (4), the first defecator (5), the first resin column (6), the second resin column (7), the second defecator (8), falling film concentrating device (9), the 3rd defecator (10)；Described the first defecator (5) is positioned at the upstream of described first resin column (6), the downstream of this first resin column (6) is described the second resin column (7), described second defecator (8) is positioned at the downstream of this second resin column (7), and described falling film concentrating device (9) is positioned at the downstream of this second defecator (8)；Described 3rd defecator (10) is positioned at the downstream of this falling film concentrating device (9)；Described centrifuge (4) has many groups, respectively in the upstream of described the first defecator (5), the upstream of the 3rd described defecator (10) and the downstream of described falling film concentrating device (9)；
As preferably, described the first defecator (5) is ultrafilter, and described second defecator (8) is collecting and filtering apparatus, and described defecator (10) is titanium rod defecator, forms by organizing titanium rod filter element in this titanium rod defecator more.
As preferably, described the first resin column (6) is positive resin post, and described the second resin column (7) is negative resin post, and described centrifuge (4) is butterfly centrifugal machine.
As preferably, described production line has three, and between the resin column (6) in every production line, pipeline is connected in parallel.
As preferably, described basin (3) has many groups, is respectively placed in described reactor (1), the first defecator (5), the first resin column (6), the second resin column (7), the second defecator (8), falling film concentrating device (9), the downstream of the 3rd defecator (10).
The process of a kind of Enzyme catalyzed synthesis AA 2G as claimed in claim 1, is characterized by: comprise the following steps:
Step one: enzyme reaction
(1) in reactor (1), add drinking water, add vitamin C, open stirring；
(2) temperature 37 DEG C in hot water controls described reactor (1), adjusts pH to 5.50 ± 0.10 with 30%NaOH solution；
(3) in described reactor (1), beta-schardinger dextrin-is added；
(4) add cyclodextrin glycosyl transferases, add drinking water and make system quantitative；
(5) this reactor (1) interior temperature in the kettle 37 ± 2 DEG C, lucifuge reaction 10-12h are controlled；
(6), after reaction 1 end, saccharifying enzyme (enzyme liquid 750,000 U/ml), temperature control 37 ± 2 DEG C, lucifuge reaction 10-14h are added；
Step 2: prepared by AA 2G crude product
(1) react complete, blowing, with drinking water washing reaction still (1), reactant liquor and kettle washing water are merged, weighs, and be centrifuged by centrifuge (4), again wash centrifuge (4) with drinking water；
(2) filtrate obtained is carried out ultrafiltration by the first defecator (5), ultra-filtration process adds drinking water, and washes with drinking water top, control temperature less than 30 DEG C；
(3) by ultrafiltrate by the first resin column, resin eluent is adsorbed in the second resin column；
(4) rinse the first resin column with drinking water, and eluent is adsorbed in the second resin column；
(5) with NaCl solution eluting resin, by eluent by the first resin column, and it is eluted to product-free outflow with drinking water, merges all eluents；
(6) the eluent merged by the second defecator, then wash with drinking water, finally concentrate, wash with drinking water top；Nanofiltration liquid is transferred in reactor (1), reduce pressure during temperature≤50 DEG C, vacuum≤-0.1MPa, concentrate, being concentrated into AA 2G content is 55%-60%；
(7) in concentrated solution, methanol, stirring and dissolving are added；Then it is cooled to 10-15 DEG C, and in 10-15 insulated and stirred 4-5 hour, continues at 1-2 hour and cool to 0-5 DEG C, then at 0-5 DEG C of insulated and stirred 4-5 hour；
(8) crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless, washs with methanol, is centrifuged after flowing out to dripless, continues centrifugal, obtains AA 2G crude product；
Step 3: AA 2G crude product refining
(1) in reactor (1), add purified water, add AA 2G crude product, open stirring；
Hot water controls temperature 35-45 DEG C in still, stirring and dissolving, adds activated carbon, insulated and stirred 30min；
(2) by solution in reactor (1) while hot by the 3rd defecator, filtrate is weighed, censorship AA 2G content；
(3) AA 2G filtrate is added in reactor (1), concentrating under reduced pressure, temperature control≤50 DEG C, vacuum≤-0.09Mpa；Being concentrated into AA 2G content in solution is 67% ± 1%；
(4) concentrating complete, control still temperature 38-45 DEG C, add crystal seed, stirring, then stand slow crystallize, slow cooling 5h, after temperature is down to 35 DEG C, programmed cooling, to 5-10 DEG C, opens stirring, then 5-10 DEG C of insulation crystallize；
(5) crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless and sprays drip washing with frozen water, continues centrifugal, is centrifuged after flowing out to dripless and continues centrifugal 30min, and discharging obtains AA 2G fine work wet product；
(6) centrifugal complete gained AA 2G wet product is placed in vacuum drying oven and is dried, temperature control≤50 DEG C, vacuum ＜-0.1Mpa；It is dried complete, obtains AA 2G fine work.
The Advantageous Effects of the present invention:
The present invention provides the process system of a kind of Enzyme catalyzed synthesis AA 2G, use ultrafilter can remove the enzyme in reactant liquor, use positive resin post can remove sodium ion, with negative resin post adsorption production, concentrate through nanofiltration through eluting, eluent again, can molecular cut off be the products molecule of 300-400.The present invention uses collecting and filtering apparatus, can remove the impurity such as vitamin C, glucose.The present invention avoids using high temperature method for concentration, and due to long-time high temperature, product can be degraded affects quality, and the present invention uses falling film concentrating device, can rapid concentration at a lower temperature, it is ensured that product quality.The present invention uses methanol plus water mixed solvent to use program crystallize, the AA 2G crystal habit stable and uniform of precipitation as AA 2G crystallization solvent, Crystallization Process, and impurity is few, and product purity is high.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of the present invention.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are merely to illustrate the present invention rather than limit the scope of the present invention.
Embodiment 1
Such as Fig. 1, the process system of a kind of Enzyme catalyzed synthesis AA 2G, is characterized by: include reactor 1, enzyme basin 2, basin 3, centrifuge the 4, first defecator the 5, first resin column the 6, second resin column the 7, second defecator 8, falling film concentrating device the 9, the 3rd defecator 10, discharging opening 11；The first described defecator 5 is positioned at the described upstream of the first resin column 6, the downstream of this first resin column 6 is the second described resin column 7, described second defecator 8 is positioned at the downstream of this second resin column 7, and described falling film concentrating device 9 is positioned at the downstream of this second defecator 8；Described 3rd defecator 10 is positioned at the downstream of this falling film concentrating device 9；Described centrifuge 4 has many groups, respectively in the upstream of the first described defecator 5, the upstream of the 3rd described defecator 10 and the downstream of described falling film concentrating device 9；
In the present embodiment, the first defecator 5 uses ultrafilter, described second defecator 8 to use collecting and filtering apparatus, described defecator (10) to be titanium rod defecator, forms by organizing titanium rod filter element in this titanium rod defecator more.Described the first resin column (6) is positive resin post, and described the second resin column (7) is negative resin post, and described centrifuge (4) is butterfly centrifugal machine.Described production line has three, and between the resin column (6) in every production line, pipeline is connected in parallel.Described basin (3) has many groups, is respectively placed in described reactor (1), the first defecator (5), the first resin column (6), the second resin column (7), the second defecator (8), falling film concentrating device (9), the downstream of the 3rd defecator (10).
Embodiment 2
In 1000L reactor, add drinking water 20KG, add vitamin C 6.5KG, stirring；By temperature 37 DEG C in hot water control reactor；PH to 5.4 is adjusted with 30%NaOH solution；In reactor, add beta-schardinger dextrin-14KG, add CGT enzyme 4.0KG, add drinking water and make system quantitatively to 65KG；Control reactor temperature 37 ± 2 DEG C, lucifuge reaction 10h；Add saccharifying enzyme (enzyme liquid 750,000 U/ml) 70g, temperature control 37 ± 2 DEG C, lucifuge reaction 10h.
React complete, blowing, with drinking water 2.0KG washing reaction still, reactant liquor and kettle washing water are merged, weighs, and be centrifuged by butterfly centrifugal machine, wash centrifuge with drinking water 2.0KG；The filtrate obtained is carried out ultrafiltration by ultrafilter, ultra-filtration process adds drinking water 30.0KG ultrafiltration, and washes with drinking water 2.0KG top, control temperature less than 30 DEG C；By ultrafiltrate by positive resin post, positive resin eluent is adsorbed in negative resin post；Rinse positive resin post with drinking water, and eluent is adsorbed in negative resin post；Use 0.03mol/L
Sodium chloride solution 100.0KG eluting negative resin；Use 0.2mol/L
Sodium chloride solution 100.0KG eluting negative resin, by eluent by positive resin post, and is eluted to product-free outflow with drinking water 100.0KG, merges all eluents；The eluent merged by collecting and filtering apparatus, then wash with drinking water 30.0KG, be finally concentrated to 20KG, wash with drinking water 2.0KG top；Transferring in reactor by nanofiltration liquid, temperature reduces pressure when 50 DEG C, concentrates during vacuum≤-0.1MPa, and being concentrated into product content is 55%；Adding methanol 30.0KG, stirring and dissolving in concentrated solution, be then cooled to 10 DEG C, and in 10 DEG C of insulated and stirred 4 hours, be cooled to 0 DEG C, insulated and stirred carries out crystallize in 4 hours；Crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless, washs with methanol 5.0KG, is centrifuged after flowing out to dripless, continues centrifugal 30min, obtains AA 2G wet product 4.0KG.
In 20L reactor, add purified water 8.0KG, add AA 2G wet product 4.0KG, open stirring；By temperature 40 DEG C in hot water control reactor, stirring and dissolving, add activated carbon 60.0g, insulated and stirred 30min；By solution in reactor while hot by titanium rod defecator, AA 2G filtrate is added in 20L falling film concentrating device, concentrating under reduced pressure, temperature control≤50 DEG C, vacuum-0.09MPa；Being concentrated into AA 2G content in solution is 67% ± 1%；Concentrate complete, control temperature 40 DEG C in falling film concentrating device, add 25g crystal seed, stir 0.5min, then stand slow crystallize, after temperature is down to 35 DEG C, programmed cooling to 5 DEG C, open stirring, then 5 DEG C of insulation crystallize 1h；Crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless with the spraying drip washing of 0.25KG frozen water, continues centrifugal, be centrifuged after flowing out to dripless and continue to be centrifuged 30min, and discharging obtains AA 2G fine work wet product；AA 2G fine work wet product is placed in vacuum drying oven and is dried, temperature control≤50 DEG C, vacuum-0.09MPa, it is dried complete, obtains AA 2G fine work 1.65KG.
Embodiment 3
In 1000L reactor, add drinking water 20KG, add vitamin C 6.5KG, stirring；By temperature 37 DEG C in hot water control reactor；PH to 5.4 is adjusted with 30%NaOH solution；In reactor, add beta-schardinger dextrin-14KG, add CGT enzyme 4.0KG, add drinking water and make system quantitatively to 65KG；Control reactor temperature 37 ± 2 DEG C, lucifuge reaction 10h；Add saccharifying enzyme (enzyme liquid 750,000 U/ml) 70g, temperature control 37 ± 2 DEG C, lucifuge reaction 14h.
React complete, blowing, with drinking water 2.0KG washing reaction still, reactant liquor and kettle washing water are merged, weighs, and be centrifuged by butterfly centrifugal machine, wash centrifuge with drinking water 2.0KG；The filtrate obtained is carried out ultrafiltration by ultrafilter, ultra-filtration process adds drinking water 30.0KG ultrafiltration, and washes with drinking water 2.0KG top, control temperature less than 30 DEG C；By ultrafiltrate by positive resin post, positive resin eluent is adsorbed in negative resin post；Rinse positive resin post with drinking water, and eluent is adsorbed in negative resin post；Use 0.03mol/L
Sodium chloride solution 100.0KG eluting negative resin；Use 0.2mol/L
Sodium chloride solution 100.0KG eluting negative resin, by eluent by positive resin post, and is eluted to product-free outflow with drinking water 100.0KG, merges all eluents；The eluent merged by collecting and filtering apparatus, then wash with drinking water 30.0KG, be finally concentrated to 40KG, wash with drinking water 2.0KG top；Transferring in reactor by nanofiltration liquid, temperature reduces pressure when 50 DEG C, concentrates during vacuum≤-0.01MPa, and being concentrated into product content is 60%；Adding methanol 30.0KG, stirring and dissolving in concentrated solution, be then cooled to 10 DEG C, and in 10 DEG C of insulated and stirred 4 hours, be cooled to 5 DEG C, insulated and stirred carries out crystallize in 5 hours；Crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless, washs with methanol 5.0KG, is centrifuged after flowing out to dripless, continues centrifugal 30min, obtains AA 2G wet product 4.0KG.
In 20L reactor, add purified water 8.0KG, add AA 2G wet product 4.0KG, open stirring；By temperature 40 DEG C in hot water control reactor, stirring and dissolving, add activated carbon 60.0g, insulated and stirred 30min；By solution in reactor while hot by titanium rod defecator, AA 2G filtrate is added in 20L falling film concentrating device, concentrating under reduced pressure, temperature control≤50 DEG C, vacuum-0.09MPa；Being concentrated into AA 2G content in solution is 67% ± 1%；Concentrate complete, control temperature 40 DEG C in falling film concentrating device, add 25g crystal seed, stir 0.5min, then stand slow crystallize, after temperature is down to 35 DEG C, programmed cooling to 10 DEG C, open stirring, then 5 DEG C of insulation crystallize 1h；Crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless with the spraying drip washing of 0.25KG frozen water, continues centrifugal, be centrifuged after flowing out to dripless and continue to be centrifuged 30min, and discharging obtains AA 2G fine work wet product；AA 2G fine work wet product is placed in vacuum drying oven and is dried, temperature control≤50 DEG C, vacuum≤-0.09MPa, it is dried complete, obtains AA 2G fine work 2.0KG.
Embodiment 4
In 1000L reactor, add drinking water 20KG, add vitamin C 6.5KG, stirring；By temperature 37 DEG C in hot water control reactor；PH to 5.4 is adjusted with 30%NaOH solution；In reactor, add beta-schardinger dextrin-14KG, add CGT enzyme 4.0KG, add drinking water and make system quantitatively to 65KG；Control reactor temperature 37 ± 2 DEG C, lucifuge reaction 11h；Add saccharifying enzyme (enzyme liquid 750,000 U/ml) 70g, temperature control 37 ± 2 DEG C, lucifuge reaction 12h.
React complete, blowing, with drinking water 2.0KG washing reaction still, reactant liquor and kettle washing water are merged, weighs, and be centrifuged by butterfly centrifugal machine, wash centrifuge with drinking water 2.0KG；The filtrate obtained is carried out ultrafiltration by ultrafilter, ultra-filtration process adds drinking water 30.0KG ultrafiltration, and washes with drinking water 2.0KG top, control temperature less than 30 DEG C；By ultrafiltrate by positive resin post, positive resin eluent is adsorbed in negative resin post；Rinse positive resin post with drinking water, and eluent is adsorbed in negative resin post；Use 0.03mol/L
Sodium chloride solution 100.0KG eluting negative resin；Use 0.2mol/L
Sodium chloride solution 100.0KG eluting negative resin, by eluent by positive resin post, and is eluted to product-free outflow with drinking water 100.0KG, merges all eluents；The eluent merged by collecting and filtering apparatus, then wash with drinking water 30.0KG, be finally concentrated to 35KG, wash with drinking water 2.0KG top；Transferring in reactor by nanofiltration liquid, temperature reduces pressure when 50 DEG C, concentrates during vacuum≤-0.01MPa, and being concentrated into product content is 60%；Adding methanol 30.0KG, stirring and dissolving in concentrated solution, be then cooled to 10 DEG C, and in 10 DEG C of insulated and stirred 4 hours, be cooled to 8 DEG C, insulated and stirred carries out crystallize in 5 hours；Crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless, washs with methanol 5.0KG, is centrifuged after flowing out to dripless, continues centrifugal 30min, obtains AA 2G wet product 4.0KG.
In 20L reactor, add purified water 8.0KG, add AA 2G wet product 4.0KG, open stirring；By temperature 40 DEG C in hot water control reactor, stirring and dissolving, add activated carbon 60.0g, insulated and stirred 30min；By solution in reactor while hot by titanium rod defecator, AA 2G filtrate is added in 20L falling film concentrating device, concentrating under reduced pressure, temperature control 48 DEG C, vacuum-0.09MPa；Being concentrated into AA 2G content in solution is 67% ± 1%；Concentrate complete, control temperature 40 DEG C in falling film concentrating device, add 25g crystal seed, stir 0.5min, then stand slow crystallize, after temperature is down to 38 DEG C, programmed cooling to 6 DEG C, open stirring, then 7 DEG C of insulation crystallize 1h；Crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless with the spraying drip washing of 0.25KG frozen water, continues centrifugal, be centrifuged after flowing out to dripless and continue to be centrifuged 30min, and discharging obtains AA 2G fine work wet product；AA 2G fine work wet product is placed in vacuum drying oven and is dried, temperature control 48 DEG C, vacuum-0.09MPa, it is dried complete, obtains AA 2G fine work 1.50KG.
Present specification describes some embodiments, it should be understood, however, that those skilled in the art would know that the various improvement without departing substantially from the spirit and scope of the present invention by reading this specification.Therefore, these other embodiments should also be as being included in scope.
Claims (1)
1. a process for Enzyme catalyzed synthesis AA 2G, uses following device: reactor (1), enzyme basin (2), basin (3), centrifuge (4), the first defecator (5), the first resin column (6), the second resin column (7), the second defecator (8), falling film concentrating device (9), the 3rd defecator (10)；Described the first defecator (5) is positioned at the upstream of described first resin column (6), the downstream of this first resin column (6) is described the second resin column (7), described second defecator (8) is positioned at the downstream of this second resin column (7), and described falling film concentrating device (9) is positioned at the downstream of this second defecator (8)；Described 3rd defecator (10) is positioned at the downstream of this falling film concentrating device (9)；Described centrifuge (4) has many groups, respectively in the upstream of described the first defecator (5), the upstream of the 3rd described defecator (10) and the downstream of described falling film concentrating device (9)；Described the first defecator (5) is ultrafilter, and described second defecator (8) is collecting and filtering apparatus, and described 3rd defecator (10) is titanium rod defecator, forms by organizing titanium rod filter element in this titanium rod defecator more；Described the first resin column (6) is positive resin post, and described the second resin column (7) is negative resin post；Described centrifuge (4) is butterfly centrifugal machine；Described basin (3) has many groups, it is respectively placed in described reactor (1), the first defecator (5), the first resin column (6), the second resin column (7), the second defecator (8), falling film concentrating device (9), the downstream of the 3rd defecator (10), it is characterized by: comprise the following steps:
Step one: enzyme reaction
(1) in reactor (1), add drinking water, add vitamin C, open stirring；
(2) temperature 37 DEG C in hot water controls described reactor (1), adjusts pH to 5.50 ± 0.10 with 30%NaOH solution；
(3) in described reactor (1), beta-schardinger dextrin-is added；
(4) add cyclodextrin glycosyl transferases, add drinking water and make system quantitative；
(5) this reactor (1) interior temperature in the kettle 37 ± 2 DEG C, lucifuge reaction 10-12h are controlled；
(6) after reaction 1 end, adding saccharifying enzyme, the concentration of this saccharifying enzyme is 750,000 U/ml, temperature control 37 ± 2 DEG C, lucifuge reaction 10-14h；
Step 2: prepared by AA 2G crude product
(1) react complete, blowing, with drinking water washing reaction still (1), reactant liquor and kettle washing water are merged, weighs, and be centrifuged by centrifuge (4), again wash centrifuge (4) with drinking water；
(2) filtrate obtained is carried out ultrafiltration by the first defecator (5), ultra-filtration process adds drinking water, and washes with drinking water top, control temperature less than 30 DEG C；
(3) by ultrafiltrate by the first resin column (6), eluent is adsorbed in the second resin column (7)；
(4) rinse the first resin column (6) with drinking water, and eluent is adsorbed in the second resin column (7)；
(5) with sodium chloride solution eluting resin, by eluent by the first resin column (6), and it is eluted to product-free outflow with drinking water, merges all eluents；
(6) the eluent merged by the second defecator (8), then wash with drinking water, finally concentrate, wash with drinking water top；Being transferred to by nanofiltration liquid in reactor (1), reduce pressure during temperature≤50 DEG C, concentrate during vacuum≤-0.1MPa, being concentrated into AA 2G content is 55%-60%；
(7) in concentrated solution, methanol, stirring and dissolving are added；Then it is cooled to 10-15 DEG C, and in 10-15 insulated and stirred 4-5 hour, continues at 1-2 hour and cool to 0-5 DEG C, then at 0-5 DEG C of insulated and stirred 4-5 hour；
(8) crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless, washs with methanol, is centrifuged after flowing out to dripless, continues centrifugal, obtains AA 2G crude product；
Step 3: AA 2G crude product refining
(1) in reactor (1), add purified water, add AA 2G crude product, open stirring；
Hot water controls temperature 35-45 DEG C in still, stirring and dissolving, adds activated carbon, insulated and stirred 30min；
(2) by solution in reactor (1) while hot by the 3rd defecator (10), filtrate is weighed, censorship AA 2G content；
(3) AA 2G filtrate is added in reactor (1), concentrating under reduced pressure, temperature control≤50 DEG C, vacuum >=-0.09Mpa；Being concentrated into AA 2G content in solution is 67% ± 1%；
(4) concentrating complete, control still temperature 38-45 DEG C, add crystal seed, stirring, then stand slow crystallize, slow cooling 5h, after temperature is down to 35 DEG C, programmed cooling, to 5-10 DEG C, opens stirring, then 5-10 DEG C of insulation crystallize；
(5) crystallize is complete, and blowing is centrifuged, and is centrifuged after flowing out to dripless and sprays drip washing with frozen water, continues centrifugal, is centrifuged after flowing out to dripless and continues centrifugal 30min, and discharging obtains AA 2G fine work wet product；
(6) centrifugal complete gained AA 2G wet product is dried, temperature control≤50 DEG C, vacuum ＜-0.1Mpa；It is dried complete, obtains AA 2G fine work.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510063451.4A CN104560707B (en) | 2015-02-06 | 2015-02-06 | The process of a kind of Enzyme catalyzed synthesis AA 2G and process system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510063451.4A CN104560707B (en) | 2015-02-06 | 2015-02-06 | The process of a kind of Enzyme catalyzed synthesis AA 2G and process system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104560707A CN104560707A (en) | 2015-04-29 |
| CN104560707B true CN104560707B (en) | 2016-08-24 |
Family
ID=53077789
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510063451.4A Active CN104560707B (en) | 2015-02-06 | 2015-02-06 | The process of a kind of Enzyme catalyzed synthesis AA 2G and process system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104560707B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104817606A (en) * | 2015-05-02 | 2015-08-05 | 江苏诚信药业有限公司 | Process system for preparing telbivudine |
| CN105018551A (en) * | 2015-07-10 | 2015-11-04 | 常州市宏硕电子有限公司 | Production system of L-ascorbic acid-glucoside and production technology |
| CN105906688A (en) * | 2016-05-01 | 2016-08-31 | 江苏诚信药业有限公司 | Technological system and technological method for extracting glutathione |
| CN108440611A (en) * | 2018-04-18 | 2018-08-24 | 山东众山生物科技有限公司 | A kind of vitamin C glucoside purifying process |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101397286A (en) * | 2008-11-18 | 2009-04-01 | 江苏江山制药有限公司 | Method for continuous crystallisation of vitamin C |
| CN102391101A (en) * | 2011-09-21 | 2012-03-28 | 石家庄开发区德赛化工有限公司 | Process for refining gulonic acid |
| CN103755756A (en) * | 2014-02-11 | 2014-04-30 | 江苏诚信制药有限公司 | Process system and process method for preparation of vitamin C ascorbyl glucoside |
| CN103923136A (en) * | 2014-04-20 | 2014-07-16 | 厦门世达膜科技有限公司 | Production method of ascorbyl glucoside |
-
2015
- 2015-02-06 CN CN201510063451.4A patent/CN104560707B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101397286A (en) * | 2008-11-18 | 2009-04-01 | 江苏江山制药有限公司 | Method for continuous crystallisation of vitamin C |
| CN102391101A (en) * | 2011-09-21 | 2012-03-28 | 石家庄开发区德赛化工有限公司 | Process for refining gulonic acid |
| CN103755756A (en) * | 2014-02-11 | 2014-04-30 | 江苏诚信制药有限公司 | Process system and process method for preparation of vitamin C ascorbyl glucoside |
| CN103923136A (en) * | 2014-04-20 | 2014-07-16 | 厦门世达膜科技有限公司 | Production method of ascorbyl glucoside |
Non-Patent Citations (2)
| Title |
|---|
| 维生素C葡萄糖苷的研究现状及发展前景;王冰等;《生物加工过程》;20080731;第6卷(第4期);1-5 * |
| 降膜蒸发器与维C工业;赵元军;《化工设备设计》;19961231;第33卷;2.1节 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104560707A (en) | 2015-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104560707B (en) | The process of a kind of Enzyme catalyzed synthesis AA 2G and process system | |
| CN104529755B (en) | A kind of method being separated α-ketoglutaric acid from conversion fluid | |
| CN106188167A (en) | A kind of separation and Extraction N acetyl group D glucosamine and method of D glucosamine from D-glucosamine fermentation liquid | |
| CN102702274A (en) | Method for preparing high-purity soybean oligosaccharide from soybean whey wastewater | |
| CN108440624B (en) | Environment-friendly method for separating and purifying cytidine from microbial fermentation broth | |
| CN105294467A (en) | Novel process for extracting feed-grade valine from fermentation liquid | |
| CN106800586A (en) | A kind of method of Moringa protein high efficiency extraction | |
| CN103923136A (en) | Production method of ascorbyl glucoside | |
| US20230167474A1 (en) | Methods for enzymatic production of glucosamine salts and the purification methods thereof | |
| CN104744523B (en) | Method for preparation of D-fructose and D-mannose by D-glucose isomerization | |
| CN106591384A (en) | Comprehensive treatment method of xylose mother liquor | |
| CN104561202A (en) | Preparation method and technological system for enzymatically synthesizing N(2)-L-alanyl-L-glutamine | |
| CN103113440A (en) | Preparation method of erythromycin thiocyanate | |
| CN111171097A (en) | Separation and purification method for producing adenosine by fermentation | |
| US20220081732A1 (en) | Process for making l-fucose | |
| CN109553645B (en) | Method for extracting low-content erythromycin A in fermentation solution | |
| CN104591999A (en) | Long chain organic acid purifying method | |
| CN111892498A (en) | Method for extracting L-malic acid | |
| CN103539688B (en) | A kind of method of separation and Extraction Serine from Corynebacterium glutamicum fermented liquid | |
| CN204474687U (en) | A kind of process system of Enzyme catalyzed synthesis AA 2G | |
| CN108640816B (en) | Refining process for inositol filtrate by high-temperature rapid ion exchange | |
| CN116283553A (en) | Method for extracting 3, 4-dihydroxybenzoic acid from fermentation broth | |
| CN105232457A (en) | Method for preparing doramectin sterile solution | |
| CN114573644A (en) | Preparation method of sialic acid | |
| CN104152508A (en) | Method for direct extraction of genipin from waste liquid in gardenia yellow pigment production |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |