CN108559767A - A kind of method that mixed pantoic acid lactone is continuously split and its device - Google Patents
A kind of method that mixed pantoic acid lactone is continuously split and its device Download PDFInfo
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Abstract
The present invention provides a kind of method that mixed pantoic acid lactone is continuously split, to splitting, addition splits enzyme in column and mixed pantoic acid lactone aqueous solution is split, after fractionation reaches balance, continuous conveying mixed pantoic acid lactone aqueous solution is continuously split since splitting column bottom, solution after balance is then flowed out from fractionation top end, the rate A of continuous conveying mixed pantoic acid lactone aqueous solution meets following condition, in formula：A is rate, and unit L/min, V are to split column volume, unit L；T is to split the mixed pantoic acid lactone of column volume to reach balance setting used and split time, unit h；60 indicate 60min/h.The continuous fractionation of mixed pantoic acid lactone may be implemented in a kind of method that mixed pantoic acid lactone is continuously split provided by the invention and its device, and can save place while improving efficiency reduces cost.
Description
Technical field
The present invention relates to a kind of enzymes to split technical field, and in particular to a kind of method that mixed pantoic acid lactone is continuously split
And its device.
Background technology
The a member of D-VB5 calcium as B family vitamin is widely used in feed addictive, food additives and medicine neck
Domain, indispensable especially in animal feed additive, demand rises year by year, is even more to form the office that supply falls short of demand in recent years
Face, price constantly raise up.
In the production of D-VB5 calcium, Pantothenic acid (chemical name：(+) 2,4- dihydroxy -3,3- acid dimethyls) be
Key intermediate.Chemical method synthesize D-VB5 technical maturity, but complex steps and to racemization product carry out crystallization fractionation expense
It is expensive.Synthesis technology can further be upgraded by producing optically pure D-VB5 using biological enzyme, biological enzyme mainly including the use of
Biomass fermenting and producing and enzyme law catalysis.Biomass fermentation method is using technique for gene engineering in Escherichia coli, glutamic acid rod
Pantothenate biosynthetic gene is expressed in the host strains such as shape bacillus, bacillus subtilis and saccharomyces cerevisiae and amino acid bio synthesizes base
Cause, and microbial fermentation is carried out using genetic engineering bacterium, Pantothenate Production is higher, but microbe fermentation method fermentation broth contents are complicated,
Subsequent product separation and Extraction is more difficult.Enzyme law catalysis can substitute splitting step in chemical reaction process, and microbial enzyme (string
Pearl Fusarium) Hydrolysis Resolution technology can obtain very high optical purity, and it is environmental-friendly, and be used widely, but according to
It is old to there is the problems such as fractionation cost is higher and fractionation is less efficient.
No matter which kind of mode is used at present, it is to use batch production to split present situation, that is, uses relatively large container (logical
Frequently with geosyncline), DL- esters (pantoic acid lactone of mixed) are prepared into certain density aqueous solution, put into a certain amount of fractionation enzyme,
The some time is stirred, during which adjusts pH value with ammonium hydroxide, the hydrolyzate of fractionation, which reaches after optical activity requires, to be filtered, and obtained mixing is molten
Liquid is made of DL- esters, L- esters and D- acid；It then stops operation, exports hydrolyzate, refill new DL- esters, repeat above-mentioned behaviour
Make, splits inefficiency.
Invention content
The present invention in view of the deficiency of the prior art, provides a kind of method that mixed pantoic acid lactone is continuously split
And its device, the continuous fractionation of mixed pantoic acid lactone may be implemented, can save place while improving efficiency reduces cost.
The technical solution adopted in the present invention：A kind of method that mixed pantoic acid lactone is continuously split adds to splitting in column
Enter to split enzyme and mixed pantoic acid lactone aqueous solution is split, it is continuous defeated since splitting column bottom after fractionation reaches balance
Mixed pantoic acid lactone aqueous solution is sent continuously to be split, for the solution after balance then from top end outflow is split, continuous conveying is mixed
The rate A of rotation pantoic acid lactone aqueous solution meets following condition：
In formula：A is rate, and unit L/min, V are to split column volume, unit L；T is to split column volume
Mixed pantoic acid lactone reaches balance setting used and splits time, unit h；60 indicate 60min/h.
Further, the addition G for enzyme being split in the present invention meets following condition：
G is the addition for splitting enzyme, unit g；V is to split column volume, unit L；C is mixed pantoic acid lactone concentration,
Unit is g/ml；The mixed pantoic acid lactone solution that T is fractionation volume V reaches balance setting used and splits time, unit h；η
For the percent hydrolysis for splitting when reaching balance, the amount/initial input DL- pantoic acid lactones for the Pantothenic acid being specially hydrolyzed into it is total
Amount；X is enzyme activity, and unit is μm ol/ming；130 be D-pantoyl lactone molecular weight.
The T of the present invention, that is, the mixed pantoic acid lactone solution for splitting volume V reach the balance setting fractionation time used,
The time generally independently set, can according to split the size of column, fractionation amount number independently selected, generally in 24-36h
Between, such as 24 hours, 36h etc..
In split process, with the generation of Pantothenic acid, pH value can decline therewith, in order to ensure that the fractionation for splitting enzyme is lived
Property, the preferred control ph of split process of the present invention is 5.2-8.8, preferably 6-7.5, more preferably 6.5-7.1；Inventor has found
It within the scope of this, can preferably ensure the activity for splitting enzyme, especially in preferred scope, split enzymatic activity and reach highest.In order to
It avoids bringing other impurities, such as metal ion into, it is ammonium hydroxide that the present invention, which adjusts the alkali used in pH,.
In order to realize that more preferable fractionation effect faster, the present invention are preferably passed through air in fractionation column and carry out bubbling stirring,
So that the fractionation enzyme in fractionation column is in fluidized state, be uniformly dispersed, to improve fractionation efficiency, ensures to split effect.
The criterion that present invention fractionation reaches balance is that solution ph is almost unchanged in fractionation column, and general pH is stable
More frequent between 6.5-7.0, pH is 6.8 or so.
In order to which the mixed pantoic acid lactone aqueous solution and the solution for having reached fractionation balance that ensure bottom conveying are incomplete
Obscure, the draw ratio (i.e. the ratio of length and diameter) that the present invention splits column needs to meet (9-15)：1, preferably (10-12.5)：
1。
The purpose continuously split may be implemented in the method for the invention, continuously logical according to special speed by splitting column bottom
Enter to revolve pantoic acid lactone aqueous solution, top outflow achievees the purpose that the solution for splitting balance continuously splits to realize；It is defeated when continuing
After sending mixed pantoic acid lactone aqueous solution, the optical activity for the solution for splitting column top outflow is detected by timing, works as optical activity
Drop to defined numerical value, generally+8 hereinafter, illustrating that the activity of enzyme is declined, can be continued by way of supplementing new enzyme
It uninterruptedly being split, after supplement is multiple, splits the accumulation due to enzyme in column, concentration increases, and sticky arrive influences mass transport process,
Enzyme can more be renewed；Invention is passed through the study found that generally when the dosage of total enzyme (be initially added plus subsequently supplement) reaches mixed
Pantoic acid lactone amount 1/4 when, need replacing new enzyme, that is, the dosage for needing to meet total enzyme is less than mixed pantoic acid lactone amount
1/4。
The device that the present invention also provides a kind of continuously to split for mixed substance, described device include split column, head tank,
Alkali liquor storage tank, the head tank is pumped by feedstock transportation to be connected with the feed inlet for splitting column bottom, and the alkali liquor storage tank passes through alkali
Liquid delivery pump is connected with the lye import for splitting column bottom；PH probes are connected at the fractionation column tube body, are split top end and are equipped with
Discharge port；The feedstock transportation pump has or is connected with flow control valve.
Further, pH probes of the present invention connect flow controller with lye delivery pump, are popped one's head in and are detected by pH
The pH value for splitting solution in column, when pH value is less than 5.2, flow controller controls lye delivery pump and supplements lye, until adjustment
PH is 5.2-8.8, stops supplement lye.Preferred adjustment pH is 6-7.5, more preferably 6.5-7.1；When pH value is almost without change
To change, general pH is stablized between 6.5-7.0, more frequent, and pH is to indicate that splitting enzymatic activity is decreased obviously at 6.8 or so,
Feedstock transportation pump can be controlled at this time to stop that raw material is added.
Further, the raw material flow rate A of flow control valve control of the present invention meets following condition
In formula：A is rate, and unit L/min, V are to split column volume, unit L；T is to split column volume
Mixed pantoic acid lactone reaches balance setting used and splits time, unit h；60 indicate 60min/h.
Further, fractionation column bottom end of the present invention connects air compressor, can be by air compressor to fractionation
Air is blasted in column and carries out bubbling stirring so that is split enzyme and is in boiling-like, is uniformly dispersed；It is preferred to split column bottom end and air
Compressor connecting place is equipped with valve, can control air-flow size by valve, so that fractionation enzyme fluidized state is reached and split pillar height degree
1/3~2/3 be preferred, can not only ensure to split efficiency, but also ensure that top has reached and split balancing liquid not in not up to flat
The liquid of weighing apparatus is obscured completely, it is ensured that has reached the outflow for splitting balancing liquid.
Further, the draw ratio (i.e. the ratio of length and diameter) of present invention fractionation column needs to meet (9-15)：1, preferably
For (10-12.5)：1, within this range, it is ensured that the mixed pantoic acid lactone aqueous solution and have reached fractionation that bottom conveys
The solution of balance is not exclusively obscured.
Further, it is ammonia in alkali liquor storage tank of the present invention in order to avoid bringing other impurities, such as metal ion into
Water.
Further, it splits to be additionally provided at column tube body and mends enzyme mouth, can be used for supplement when enzyme activity reduces and split enzyme.
Further, it splits top end and is internally provided with filter element, can prevent fractionation enzyme from being flowed out with liquid.
Further, it splits column bottom inner portion and also is provided with filter element, you can with dispersion air, and can prevent enzyme from flowing back to
Feed inlet blocks pipeline.
Further, the top for splitting column is equipped with sample tap, is used for the optical activity of sample detection solution, when optical activity declines
To defined numerical value, generally+8 hereinafter, illustrating that the activity of enzyme is declined, new enzyme can be supplemented, after supplement is multiple, is split
It is sticky to mass transport process is influenced since the accumulation of enzyme, concentration increase in column, it can more renew enzyme；Generally when the dosage of total enzyme
When (be initially added plus subsequently supplement) reaches the 1/4 of mixed pantoic acid lactone amount, new enzyme is needed replacing, that is, needs to meet total enzyme
Dosage be less than mixed pantoic acid lactone amount 1/4.
Further, the discharge port connection splits liquid storage tank, is balanced for storing the fractionation that reaches flowed out from discharge port
Liquid；It is furthermore preferred that flow instrument is additionally provided between the discharge port and fractionation liquid storage tank, for monitoring yield.
The advantage of the present invention compared with the existing technology：
1) it continuously can be passed through rotation pantoic acid lactone aqueous solution according to special speed by splitting column bottom, top outflow reaches
Continuous fractionation is realized to the solution for splitting balance, is split efficient；And it can be automatic by connecting the realizations such as flow controller
Control.
2) it is not necessarily to build to split pond, floor space is small, reduces cost；
3) present invention is by splitting bubbling stirring in column, compared with pumping forced circulation in classical resolution method, to enzyme activity
Property destructiveness smaller, can extend the service life of enzyme, in addition, being found through overtesting, in conventional batch method for splitting, when substrate is dense
Degree drops to some range (2% or less such as), can not work although enzyme is still active；And herein described method, begin
The normal operating conditions of enzyme can be kept eventually, until enzyme complete deactivation；It is found by many experiments, usually interval is split, per 24-
An enzyme will be filtered within 30 hours to feed intake again, and continuous method for splitting of the present invention can be 5 days or so just start to add it is new
Enzyme.
So-called " lactone " " DL- esters " or " DL- lactones " of the invention, is the pantoic acid lactone of mixed, i.e.,；
The present invention is so-called " D- acid ", as Pantothenic acid；
So-called " L- esters " of the invention, as L- pantoic acid lactones.
Description of the drawings
Fig. 1 is the structural schematic diagram of detachment device of the present invention；
Wherein：1 splits column, and 2 head tanks, 3 feedstock transportations pump, 4 mend enzyme mouth, and 5 split liquid storage tank, 6 flow instruments, and 7pH pops one's head in,
8 flow controllers, 9 lye delivery pumps, 10 alkali liquor storage tanks, 11 air compressors, 12 sample taps, 13 feed inlets, 14 lye imports,
15 discharge ports；16 filter elements.
Specific implementation mode
With reference to embodiment, the present invention will be further described, it should be understood that specific embodiment described herein is only
To explain the present invention, it is not intended to limit the present invention, all letters under the concept thereof of the present invention to preparation method of the present invention
Single improve belongs within protection scope of the present invention.Following example test method without specific conditions, usually according to
The known approaches of this field.
Embodiment 1
A kind of method that mixed pantoic acid lactone is continuously split splits enzyme and mixed pantoic acid lactone to splitting to be added in column
Aqueous solution is split, fractionation reach balance after, since splitting column bottom continuous conveying mixed pantoic acid lactone aqueous solution into
Row is continuous to be split, and the solution after balance is then flowed out from fractionation top end, the rate A of continuous conveying mixed pantoic acid lactone aqueous solution
Meet following condition：
In formula：A is rate, and unit L/min, V are to split column volume, unit L；T is to split column volume
Mixed pantoic acid lactone reaches balance setting used and splits time, unit h；60 indicate 60min/h.
The addition G that enzyme is split in the present embodiment meets following condition：
G is the addition for splitting enzyme, unit g；V is to split column volume, unit L；C is mixed pantoic acid lactone concentration,
Unit is g/ml；T is to split the mixed pantoic acid lactone of column volume to reach balance setting used and split time, unit
h；η is the percent hydrolysis split when reaching balance, the amount/initial input DL- pantoic acid lactones for the Pantothenic acid being specially hydrolyzed into
Total amount；X is enzyme activity, and unit is μm ol/ming；130 be D-pantoyl lactone molecular weight.
T in the present embodiment, that is, the mixed pantoic acid lactone for splitting column volume reach balance setting used and tear open
Between timesharing, the time generally independently set, can according to split the size of column, fractionation amount number independently selected, such as
For 24 hours, 36h etc..Split process can be 5.2-8.8 with control ph, preferably 6-7.5, more preferably 6.5-7.1；Reach balance
Criterion is that solution ph is almost unchanged in fractionation column, and general pH is stablized between 6.5-7.0, and more frequent, pH is
6.8 or so.It is ammonium hydroxide to adjust the alkali used in pH.
The present embodiment can also be passed through air in fractionation column and carry out bubbling stirring, and the fractionation enzyme in fractionation column is made to be in boiling
State is risen, is uniformly dispersed, to improve fractionation efficiency, ensures to split effect.
In the present embodiment, the draw ratio (i.e. the ratio of length and diameter) for splitting column needs to meet (9-15)：1, preferably
(10-12.5)：1, it can ensure the mixed pantoic acid lactone aqueous solution of bottom conveying within this range and have reached to split to put down
The solution of weighing apparatus is not exclusively obscured.
Embodiment 2
A kind of device continuously split for mixed substance as shown in Figure 1：
Described device include split column 1, head tank 2, alkali liquor storage tank 10, the head tank 2 by feedstock transportation pump 3 with tear open
The feed inlet 13 of 1 bottom of column is divided to be connected, the alkali liquor storage tank 10 passes through lye delivery pump 9 and splits the lye import of 1 bottom of column
14 are connected；Equipped with pH probes 7 on the fractionation column 1, splits 1 top of column and be equipped with discharge port 15；Feedstock transportation pump 3 have or
It is connected with flow control valve.The raw material flow rate A of flow control valve control meets following condition
In formula：A is rate, and unit L/min, V are to split column volume, unit L；T is to split column volume
Mixed pantoic acid lactone reaches balance setting used and splits time, unit h；60 indicate 60min/h.
PH probes 7 and lye delivery pump 14 described in the present embodiment connect flow controller 8, are torn open by 7 detection of pH probes
The pH value for dividing solution in column 1, when pH value is less than 5.2, flow controller 8 controls lye delivery pump 14 and supplements lye, until tune
Whole pH is 5.2-8.8, stops supplement lye.Preferred adjustment pH is 6-7.5, more preferably 6.5-7.1；When pH value is almost without change
To change, general pH is stablized between 6.5-7.0, more frequent, and pH is to indicate that splitting enzymatic activity is decreased obviously at 6.8 or so,
Feedstock transportation pump 3 can be controlled at this time to stop that raw material is added.
The present embodiment splits 1 bottom end of column and connects air compressor 11, can be roused into fractionation column 1 by air compressor 11
Enter air and carry out bubbling stirring so that splits enzyme and be in boiling-like, be uniformly dispersed；It is preferred to split column bottom end and air compressor
Connecting place be equipped with valve, can by valve control air-flow size, make fractionation enzyme fluidized state reach split pillar height degree 1/3~
2/3 is preferred, and can not only ensure to split efficiency, but also ensures that top has reached and split balancing liquid not in the liquid of not up to balance
Obscure completely, it is ensured that have reached split balancing liquid outflow.
The draw ratio (i.e. the ratio of length and diameter) that the present embodiment splits column 1 needs to meet (9-15)：1, preferably (10-
12.5)：1, within this range, it is ensured that the mixed pantoic acid lactone aqueous solution and have reached fractionation balance that bottom conveys
Solution is not exclusively obscured.
It splits to be additionally provided at 1 tube body of column and mends enzyme mouth 4, can be used for supplement when enzyme activity reduces and split enzyme.It splits inside top end
Equipped with filter element 16, it can prevent fractionation enzyme from being flowed out with liquid.It splits column bottom inner portion and also is provided with filter element 16, you can with
Dispersion air, and can prevent enzyme from flowing back to feed inlet and block pipeline.The top for splitting column 1 is equipped with sample tap 12, for sampling inspection
Survey the optical activity of solution.Discharge port 15 connection split liquid storage tank 5, for store flow out from discharge port 15 reach fractionation balance
Liquid；It is additionally provided with flow instrument 6 between discharge port 15 and fractionation liquid storage tank 5, for monitoring yield.
Mixed pantoic acid lactone to be split is configured to aqueous solution, is placed in head tank 2, ammonium hydroxide is configured to 10% water
Solution is placed in alkali liquor storage tank 10.
Using equipment described in 1 the method for embodiment and embodiment 2, in head tank 2 solution and a certain amount of fractionation
Enzyme is tuned into pulpous state, is added to and splits in column 1, is full of with material liquid, installs pH probes 7, as shown in Figure 1.Double solid line is in figure
Pipeline, real void two-wire is signal wire and control line.
Air compressor 11 is opened, being passed through air into hydrolysis column under room temperature carries out bubbling stirring, is passed through air capacity with one
A valve regulated so that split enzyme and be in boiling-like, be uniformly dispersed.
For mixed pantoic acid lactone aqueous solution under the action of splitting enzyme, it is general that D-pantoyl lactone therein is gradually hydrolyzed to D-
Solution acid, causes system pH to decline, and perceives and be transmitted to pH controllers 8 by pH probes 7, commander's ammonium hydroxide delivery pump 9 acts, by ammonia
Water is automatically stopped supplement ammonium hydroxide from being conveyed into fractionation column in tank used for storing ammonia 10 after pH value rises to suitable range.
Usual pH value is controlled in 5.5-8.5, appropriate range 6.5-7.5, best 6.8-7.2.
Through splitting, the pH value after a certain period of time, perceived from pH probes is almost unchanged or ammonium hydroxide delivery pump is more than 20 minutes nothings
Need work, it was demonstrated that the Hydrolysis Resolution in column has reached balance.
Manually opened feedstock transportation pump 3, mixed pantoic acid lactone solution is conveyed into column, conveying speed is about A (L/
min)：
It is calculated generally according to the activity (hydrolysis ability) for splitting enzyme, the concentration of preparation can make D-pantoyl lactone in T
Hydrolysis in interior (T=24-36 hours) reaches required level.
With the input of material liquid, D-pantoyl lactone concentration rises in column, splits enzyme and continues to start to work, it is general to generate D-
PH value declines after solution acid, and ammonium hydroxide is automatically replenished, until there is feed liquid outflow in top, is used as yield accumulation monitoring through flowmeter 6,
From 12 sample detection optical activity ＞+8 of sample tap.
As long as the activity of usual enzyme does not change or slight change can receive, so that it may be split with continuous always.Even
In continuous split process, the magnitude of recruitment of feed rate A homeostasis, ammonium hydroxide is also almost homeostasis.Some time (usual 120 hours, and
Under same scale, intermittent split usually is fed intake again per will filter within 24-30 hour an enzyme) after, because of enzyme activity decline, ammonium hydroxide
Supplement speed also begin to decline, from sample tap sample detection to optical activity ＜+8, require supplementation with new enzyme.
After supplementing 3-4 times, supplement is former every time is pumped into 20% or so of enzyme amount, and the total amount of enzyme reaches the 1.6- initially put into
1.8 times, the accumulation due to enzyme in column is split, concentration increases, sticky to mass transport process is influenced, and should more renew enzyme.
Claims (10)
1. a kind of method that mixed pantoic acid lactone is continuously split, it is characterised in that splitting, addition splits enzyme in column and mixed is general
Solution acid lactone aqueous solution is split, after fractionation reaches balance, the continuous conveying mixed pantoic acid lactone since splitting column bottom
Aqueous solution is continuously split, and for the solution after balance then from top end outflow is split, continuous conveying mixed pantoic acid lactone is water-soluble
The rate A of liquid meets following condition：
In formula：A is rate, and unit L/min, V are to split column volume, unit L；T is the mixed for splitting column volume
Pantoic acid lactone reaches balance setting used and splits time, unit h；60 indicate 60min/h.
2. the method that mixed pantoic acid lactone according to claim 1 is continuously split, it is characterised in that split the addition of enzyme
Amount G meets following condition：
In formula：G is the addition for splitting enzyme, unit g；V is to split column volume, unit L；C is mixed pantoic acid lactone concentration,
Unit is g/ml；T is to split the mixed pantoic acid lactone of column volume to reach balance setting used and split time, unit
h；η is the percent hydrolysis split when reaching balance, the amount/initial input DL- pantoic acid lactones for the Pantothenic acid being specially hydrolyzed into
Total amount；X is enzyme activity, and unit is μm ol/ming；130 be D-pantoyl lactone molecular weight.
3. the method that mixed pantoic acid lactone according to claim 1 is continuously split, it is characterised in that split process controls
PH value is 5.2-8.8, preferably 6-7.5, more preferably 6.5-7.1.
4. the method that mixed pantoic acid lactone according to claim 1 is continuously split, it is characterised in that used in control pH
Alkali is ammonium hydroxide；Air is passed through in fractionation column carry out bubbling stirring in split process.
5. the method that mixed pantoic acid lactone according to claim 1 is continuously split, it is characterised in that fractionation reaches balance
Criterion be split column in pH value of solution stablize between 6.5-7.0.
6. the method that mixed pantoic acid lactone according to claim 1 is continuously split, it is characterised in that split the major diameter of column
Than satisfaction (9-15)：1, preferably (10-12.5)：1.
7. a kind of device continuously split for mixed pantoic acid lactone, it is characterised in that described device includes splitting column, raw material
Tank, alkali liquor storage tank, the head tank is pumped by feedstock transportation to be connected with the feed inlet for splitting column bottom, and the alkali liquor storage tank passes through
Lye delivery pump is connected with the lye import for splitting column bottom；PH probes are connected at the fractionation column tube body, are split top end and are set
There is discharge port；The feedstock transportation pump has or is connected with flow control valve；The pH probes and lye delivery pump connection flow
Amount controller.
8. the device that mixed substance according to claim 7 is continuously split, it is characterised in that the fractionation column bottom end connection
Air compressor, the preferred column bottom end that splits are equipped with valve with air compressor connecting place；It splits inside top end and in bottom end
Portion is equipped with filter element；It splits to be additionally provided at column tube body and mends enzyme mouth.
9. the device that mixed substance according to claim 7 is continuously split, it is characterised in that the draw ratio for splitting column meets
(9-15)：1, preferably (10-12.5)：1.
10. the device that mixed substance according to claim 7 is continuously split, it is characterised in that split column top be equipped with take
Sample mouth；The discharge port connection splits liquid storage tank；It is additionally provided with flow instrument between preferred discharge port and fractionation liquid storage tank.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111455013A (en) * | 2020-05-14 | 2020-07-28 | 吴江 | Method for auxiliary resolution of pantolactone by weak base salt |
| CN111979288A (en) * | 2019-05-24 | 2020-11-24 | 重庆桑禾动物药业有限公司 | Method for continuously decomposing mixed pantolactone by enzymolysis through fixed bed device |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04335890A (en) * | 1991-05-08 | 1992-11-24 | Tanabe Seiyaku Co Ltd | Reaction process using immobilized biocatalyst and apparatus therefor |
| US6214609B1 (en) * | 1997-05-01 | 2001-04-10 | Monsanto Company | Method and apparatus for preparation of chiral beta amino acids using penicilln G acylase |
| CN101392278A (en) * | 2008-06-11 | 2009-03-25 | 济南大华广济畜牧发展有限公司 | Method for preparing D-pantolactone by microbe mixed fermentation method |
| CN101538564A (en) * | 2009-04-29 | 2009-09-23 | 河北宝利药业有限公司 | Immobilization method of D-pantoic acid lactone hydrolase |
| CN205382166U (en) * | 2016-03-06 | 2016-07-13 | 河北工业大学 | Device for producing polymyxin E by coupling fermentation and foam separation |
| CN106635787A (en) * | 2016-11-07 | 2017-05-10 | 鲁东大学 | Method for assembling stir-type fermentation tank system into bubble column reactor |
| CN207918811U (en) * | 2018-01-25 | 2018-09-28 | 江苏凌云药业股份有限公司 | One kind being used for the continuous detachment device of mixed pantoic acid lactone |
-
2018
- 2018-01-25 CN CN201810072934.4A patent/CN108559767A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04335890A (en) * | 1991-05-08 | 1992-11-24 | Tanabe Seiyaku Co Ltd | Reaction process using immobilized biocatalyst and apparatus therefor |
| US6214609B1 (en) * | 1997-05-01 | 2001-04-10 | Monsanto Company | Method and apparatus for preparation of chiral beta amino acids using penicilln G acylase |
| CN101392278A (en) * | 2008-06-11 | 2009-03-25 | 济南大华广济畜牧发展有限公司 | Method for preparing D-pantolactone by microbe mixed fermentation method |
| CN101538564A (en) * | 2009-04-29 | 2009-09-23 | 河北宝利药业有限公司 | Immobilization method of D-pantoic acid lactone hydrolase |
| CN205382166U (en) * | 2016-03-06 | 2016-07-13 | 河北工业大学 | Device for producing polymyxin E by coupling fermentation and foam separation |
| CN106635787A (en) * | 2016-11-07 | 2017-05-10 | 鲁东大学 | Method for assembling stir-type fermentation tank system into bubble column reactor |
| CN207918811U (en) * | 2018-01-25 | 2018-09-28 | 江苏凌云药业股份有限公司 | One kind being used for the continuous detachment device of mixed pantoic acid lactone |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111979288A (en) * | 2019-05-24 | 2020-11-24 | 重庆桑禾动物药业有限公司 | Method for continuously decomposing mixed pantolactone by enzymolysis through fixed bed device |
| CN111455013A (en) * | 2020-05-14 | 2020-07-28 | 吴江 | Method for auxiliary resolution of pantolactone by weak base salt |
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