CN1424402A - Niacinamide production by microbiological catalysis - Google Patents
Niacinamide production by microbiological catalysis Download PDFInfo
- Publication number
- CN1424402A CN1424402A CN 02151290 CN02151290A CN1424402A CN 1424402 A CN1424402 A CN 1424402A CN 02151290 CN02151290 CN 02151290 CN 02151290 A CN02151290 A CN 02151290A CN 1424402 A CN1424402 A CN 1424402A
- Authority
- CN
- China
- Prior art keywords
- niacinamide
- reaction
- under
- crystal
- concentration
- 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.)
- Pending
Links
Landscapes
- Pyridine Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
A process for preparing nicotinamide by microbe catalysis method features that the 3-nitrile pyridine is converted to nicotinamide by nitrile hydratase generated by propionate coryneform. Its advantages are simple process, less environmental pollution, and high purity of product.
Description
Technical field:
The invention belongs to microbial technology field.Be specifically related to a kind of microorganism catalysis method and produce niacinamide.
Background technology:
Niacinamide is the essential B-title complex VITAMIN of humans and animals, very wide in distributed in nature, more with content in the foods such as the plumule of dry yeast cereal, meat (liver and muscle), peanut, be the integral part of codehydrogenase in the organism, participate in carbohydrate, fat and proteinic metabolism.Niacinamide is a kind of anti-thick dermopathic VITAMIN in medicine, and some tetter and neurological disorder are had special efficacy.It can also add in food and the flour in order to replenish the VITAMIN of needed by human body, and its largest application areas is a fodder additives.Simultaneously, niacinamide also is the important intermediate of synthetic other medicine, agricultural chemicals.
Main production methods is at present: the ammonia oxidation of 1.3-picoline becomes niacinamide to become niacinamide with 2-methyl-5-ethylpyridine with nitric acid oxidation.Oxidation proceses of ammonia (the particularly nitric acid oxidation of 2-methyl-5-ethylpyridine) is the very high method of a kind of selectivity, but he is adventurous, in order to reduce this danger, high-quality personnel, best Infrastructure and higher technical level are essential, thereby are provided with big obstacle to industrialization and transfer of technology.At present, but also do not obtain the ammonia oxygen catalyst of industrial applications.
Summary of the invention:
Technical problem to be solved by this invention is to overcome above-mentioned weak point, designs a kind of production method of being convenient to industrialization.
The invention provides a kind of microorganism catalysis method and produce the method for niacinamide, this method is propionic acid rod bacillus or its mutagenic strain cell that contains Nitrile hydratase by fermentative production, become niacinamide with free cell method or the hydration of free cell continuous processing catalysis 3-itrile group pyridine under certain condition, then obtain highly purified niacinamide product by comparatively simple aftertreatment.
The specific embodiment of the present invention is as follows: the preparation of (1) zymogenic cells:
Carry out the cultivation of ferment-seeded earlier, the an amount of seed culture medium (10-70%) of in triangular flask or seeding tank, packing into, a certain amount of bacterial classification (1-10%) is inserted in the sterilization back, under 20-40 ℃, in the rotary shaking table (100-400rpm) shaking culture 30-120 hour, then at 50L-20m
3Its fermention medium Intake Quantity of fermentor tank be 40%-70%, the seed liquor of 2-7% is inserted in real jar of sterilization back, at air flow 1: 0.2-1: 1 (V.V.M); Mixing speed: 50-400rpm; Tank pressure 0.03-0.06MP; Temperature: 20-40 ℃.Fermented 30-200 hour;
Seed culture medium is formed: 0.5-2% glucose, 0.2-1% yeast extract paste, 0.05-0.15%Nacl, 0.05-0.3%K
2HPO
4, 0.01-0.03%MgSo
4, urea 0.1-1%, PH7.0-7.5;
Fermention medium: 1-2.5% glucose, 0.2-1% yeast extract paste, 0.1-2% urea, 0.03-0.1%K
2HPO
4, 0.03-0.1%KH
2PO
4, 0.03-0.1%MgSO
4, 2-20ppmCoCl
2, PH6.5-7.5; (2) catalytic hydration production process:
At separation factor 〉=3000kg, supernatant liquor is removed in centrifugation with fermented liquid, obtains the nitrile hydratase production cell; Cell suspension in the damping fluid of deionized water or pure water or PH7.0-8.0, is joined in the reactor, the scale dimension of fermented liquid and reaction solution volume is held in 〉=and 1%.Under 5-55 ℃ temperature, carry out enzymic catalytic reaction under the stirring velocity of 50-300rpm.When production concentration 〉=5%, begin to go out product by the ultra-flow membrane filtration system, begin to add 3-itrile group pyridine and water simultaneously, this membrane filtration system passes through by product, and reach removal protein and other impurity, participate in reaction and can make free cell return reactor again.By adjusting 3-itrile group pyridine and the flow acceleration of water and the take-off rate of product nicotinamide soln, thereby make effusive product nicotinamide soln concentration remain on the scope of setting, initial substrate concentration≤20% (w/v), final transformation efficiency is 〉=99%; (3) purification process:
The certain density niacinamide liquid that hydration reaction is obtained places under 1-20 ℃ the environment, keeps more than 2 hours.The reduction of the solubility with temperature of niacinamide and descending greatly, after insulation for some time, the mass crystallization body appears, the mixing solutions of crystal and water is separated the acquisition crystal by frozen centrifugation, crystal is carried out vacuum-drying under 35-50 ℃, the final niacinamide purity that obtains is more than 99.8%.
Perhaps adopt batch formula reaction, the flow process of batch formula reaction: with fermented liquid at separation factor 〉=3000kg, centrifugation, remove supernatant liquor, with cell suspension in the damping fluid of deionized water or pure water or PH7.0-8.0, the scale dimension of fermented liquid and reaction solution is held in 〉=and 1%, under 5-55 ℃ temperature, carry out enzymic catalytic reaction under the stirring velocity of 50-300rpm, in the reaction process, make concentration of substrate be controlled at 0.001%-20% by solid or the solution of adding 3-itrile group pyridine, reaction through 1-24h, obtain concentration at the nicotinamide soln more than 10%, its product solution is accumulated to behind the finite concentration (〉=10%), and disposable to go out a jar final transformation efficiency be 〉=99%;
To reach certain density reaction solution by centrifugal cell and the cell debris of removing, temperature maintenance is at 28-32 ℃, and the clear liquid that obtains is removed protein by membrane filtration system again, and removing under the environment that proteinic liquid places 1-20 ℃, maintenance is more than 2 hours.The reduction of the solubility with temperature of niacinamide and descending greatly, after insulation for some time, the mass crystallization body appears, the mixing solutions of crystal and water is separated the acquisition crystal by membrane filtration or frozen centrifugation, crystal is carried out vacuum-drying under 35-50 ℃, the final niacinamide purity that obtains is more than 99.8%.Be with the difference of continuous processing, by adding solid or the solution that adds 3-itrile group pyridine, concentration of substrate is controlled at more than the 10ppm, reaction through 1-24h, obtain concentration at the nicotinamide soln more than 10%, its product solution is accumulated to behind the finite concentration that (〉=10%) is disposable to go out jar, transformation efficiency 〉=99%.Also will add in the separation purification step, will reach certain density reaction solution by centrifugal cell and the cell debris of removing, remove protein by membrane filtration system again, temperature maintenance is at 28-32 ℃.All the other methods are all identical with continuous processing.
The Nitrile hydratase that the present invention uses propionic acid rod bacillus and mutagenesis bacterial classification thereof to produce changes into niacinamide with 3-itrile group pyridine, and its operating process is simple, reaction conditions is gentle, reduce environmental pollution, and it is simple to separate purification, the product purity height.
The present invention compared with prior art has following characteristics:
1) to be used for the bacterial strain that the catalysis nitrile becomes acid amides mainly be the bacterium of Rhod to prior art, and the present invention adopts is propionic acid rod bacillus and through the bacterial classification of mutagenesis.This bacterial classification obtains by separating in the soil of this contained nitrile, is propionic acid rod bacillus through preliminary evaluation.This strain enzyme-producing ability is strong, can obtain higher biomass under fermentation condition of the present invention, reaches as high as 40 mg/ml fermented liquids, thereby total enzyme of its unit volume fermented liquid is alive also higher, reaches as high as 1,000 ten thousand units per ml fermented liquids.(annotate: enzyme is lived and is defined, and the 3-itrile group pyridine of 1 microgram is converted into the enzyme amount of niacinamide in one hour, unit: microgram/hour).
2) technology of catalytic hydration use of the present invention is batch formula reaction or free cell membrane sepn successive reaction.With traditional batch formula reacting phase ratio, successive reaction has improved production efficiency greatly, has alleviated labour intensity, has improved biological enzyme agent and usage ratio of equipment; Relative fixed cells produce of the present invention has reduced technical process simultaneously, has reduced the three wastes, has reduced production cost, has reduced resistance to mass transfer in the production process and has improved production efficiency.The reaction of batch formula is low for equipment requirements, and the concentration for nicotinamide of production is than continuous processing height, and the separation in easy downstream is purified.
3) separation of the present invention is purified and is adopted the reaction solution freezing and crystallizing, and after-filtration or centrifugal, again with crystal vacuum-drying or spraying drying.
Embodiment:
Embodiment 1,
The substratum of adding 60% in the seeding tank of 1m3 after the real jar of sterilization, inserts the slant strains of 1 eggplant bottle, and the air flow of earlier fermentation is 1: 0.3, stirring velocity is 50rpm, and the air flow in later stage is 1: 0.5, and stirring velocity is 100rpm, tank pressure 0.05mp, 28 ℃ of temperature were cultivated 50 hours.
At 10m
3Fermentor tank in add 60% fermention medium, 5% seed liquor is inserted in real jar of sterilization back, at air flow 1: 0.5 (V.V.M); Tank pressure 0.05MP; Mixing speed: the early stage 180rpm, later stage 100rpm; Temperature: 28 ℃.Fermented 60 hours.
Seed culture medium is formed: 0.6% glucose, 0.4% yeast extract paste, 0.05%K
2HPO
4, 0.05%MgSo
4, 0.05%KH
2PO
4, PH7.2
Fermention medium: 2.1% glucose, 0.4% yeast extract paste, 0.9% urea, 0.05%K
2HPO
4, 0.05%KH
2PO
4, 0.1%MgSO
4, 10ppm CoCl
2, PH7.5
Embodiment 2,
With 50 milliliters of fermented liquids when separation factor is 3500kg, centrifugal 7 minutes.Remove supernatant liquor, obtain the cell of nitrile hydratase production.In deionized water solution, the ratio of fermented liquid and reaction solution volume is 5% with cell suspension.Under 28 ℃ temperature, carry out enzymic catalytic reaction under the stirring velocity of 100rpm.The starting point concentration of 3-itrile group pyridine is 5%, preceding 30min adds the 3-itrile group pyridine solution of concentration 50% with the flow velocity of 15ml/min, and after product nicotinamide soln (about 20%) passes through ultra flow membrane filtration system with the speed of 1000ml/h, enter down technology one, simultaneously 16.5% 3-itrile group pyridine solution adds with the velocity flow of 1030ml/h and enters the continuous reaction jar.Termination reaction after 96 hours has the 3-itrile group pyridine of 19280g to be converted into niacinamide, and transformation efficiency is 〉=99%.
Embodiment 3,
To place from the reaction solution that successive reaction obtains under 4 ℃ the freezing environment, keep more than 2 hours.The reduction of the solubility with temperature of niacinamide and descending greatly after freezing insulation for some time, the mass crystallization body occurs.The mixing solutions of crystal and water is separated the acquisition crystal by frozen centrifugation.Crystal is carried out vacuum-drying under 40 ℃, the final niacinamide purity that obtains is more than 99.8%.
Embodiment 4,
Is 3500kg with 1 liter of fermented liquid at separation factor, centrifugal 7 minutes.Remove supernatant liquor, obtain the cell of 20g.In deionized water, the scale dimension of fermented liquid and reaction liquid is held in 5% (v/v) with cell suspension.Under 28 ℃ temperature, carry out enzymic catalytic reaction under the stirring velocity of 100rpm.Make concentration of substrate about 5% by adding 3-itrile group pyridine solid,, constantly add a certain amount of substrate and make concentration of substrate maintain 100ppm along with the continuation of reaction.The ending stage in reaction reduces concentration of substrate thereby no longer add substrate gradually, trends towards zero.Have the cigarette nitrile of 6000g to be converted into niacinamide after one hour, the nicotinamide soln concentration that obtains is that 30% (w/v) transformation efficiency is 〉=99%.
Embodiment 5,
To reach certain density reaction solution by centrifugal cell and the cell debris of removing, remove protein by membrane filtration then.The clear liquid that obtains placed under 4 ℃ the freezing environment, keep more than 2 hours.The reduction of the solubility with temperature of niacinamide and descending greatly after freezing insulation for some time, the mass crystallization body occurs.The mixing solutions of crystal and water is separated the acquisition crystal by membrane filtration or frozen centrifugation.Crystal is carried out vacuum-drying under 40 ℃, the final niacinamide purity that obtains is more than 99.8%.
Claims (2)
1, a kind of microorganism catalysis method is produced the method for niacinamide, it is characterized in that this method comprises the following steps: the preparation of (1) zymogenic cells:
Carry out the cultivation of ferment-seeded earlier, the an amount of seed culture medium (10-70%) of in triangular flask or fermentor tank, packing into, a certain amount of bacterial classification (1-10%) is inserted in the sterilization back, under 20-40 ℃, in the rotary shaking table (100-400rpm) shaking culture 48-120 hour, its fermention medium Intake Quantity of fermentor tank at 50L-20m3 is 40%-70% then, and the seed liquor of 2-7% is inserted in real jar of sterilization back, at air flow 1: 0.2-1: 1 (V.V.M); Tank pressure 0.03-0.06MP; Mixing speed: 50-400rpm; Temperature: 20-40 ℃, fermented 30-200 hour;
Seed culture medium is formed: 0.5-2% glucose, 0.2-1% yeast extract paste, 0.05-0.15%Nacl, 0.05-0.3%K2HPO4,0.01-0.03%MgSo4, urea 0.1-1%, PH7.0-7.5;
Fermention medium: 1-2.5% glucose, 0.2-1% yeast extract paste, 0.1-2% urea, 0.03-0.1%K2HPO4,0.03-0.1%KH2PO4,0.03-0.1%MgSO4,2-20ppmCoCl2, PH6.5-7.5; (2) catalytic hydration production process:
At separation factor 〉=3000kg, supernatant liquor is removed in centrifugation with fermented liquid, obtains the nitrile hydratase production cell; With cell suspension in the damping fluid of deionized water or pure water or PH7.0-8.0, join in the successive reaction jar, the scale dimension of fermented liquid and reaction liquid is held in 〉=and 1%, under 5-55 ℃ temperature, carry out enzymic catalytic reaction under the stirring velocity of 50-300rpm, when production concentration 〉=5%, begin to go out product by ultra flow membrane filtration system, begin to add 3-itrile group pyridine and water simultaneously, this membrane filtration system passes through by product, participate in reaction and make free cell return reactor again, by regulating 3-itrile group pyridine and the flow acceleration of water and the take-off rate of product nicotinamide soln, thereby make effusive product nicotinamide soln concentration remain on the scope of setting, initial substrate concentration≤20% (w/v), final transformation efficiency is 〉=99%; (3) purification process:
The certain density niacinamide liquid that hydration reaction is obtained places under 1-4 ℃ the freezing environment, keep more than 2 hours, the reduction of the solubility with temperature of niacinamide and descending greatly, after insulation for some time, the mass crystallization body appears, the mixing solutions of crystal and water is separated the acquisition crystal by frozen centrifugation, crystal is carried out vacuum-drying under 35-50 ℃, the final niacinamide purity that obtains is more than 99.8%.
2, a kind of microorganism catalysis method according to claim 1 is produced the method for niacinamide, it is characterized in that wherein said step (2) and (3) adopt following method:
With fermented liquid at separation factor 〉=3000kg, centrifugation, remove supernatant liquor, with cell suspension in the damping fluid of deionized water or pure water or PH7.0-8.0, the scale dimension of fermented liquid and reaction solution is held in 〉=and 1%, under 5-55 ℃ temperature, carry out enzymic catalytic reaction under the stirring velocity of 50-300rpm, in the reaction process, make concentration of substrate be controlled at 0.001%-20% by solid or the solution of adding 3-itrile group pyridine, through the reaction of 1-24h, obtain concentration at the nicotinamide soln more than 10%, its product solution is accumulated to behind the finite concentration (〉=10%), and disposable to go out a jar final transformation efficiency be 〉=99%;
To reach certain density reaction solution by centrifugal cell and the cell debris of removing, temperature maintenance is at 28-32 ℃, and the clear liquid that obtains is removed protein by membrane filtration system again, and removing under the environment that proteinic liquid places 1-20 ℃, maintenance is more than 2 hours.The reduction of the solubility with temperature of niacinamide and descending greatly, after freezing insulation for some time, the mass crystallization body appears, the mixing solutions of crystal and water is separated the acquisition crystal by membrane filtration or frozen centrifugation, crystal is carried out vacuum-drying under 35-50 ℃, the final niacinamide purity that obtains is more than 99.8%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02151290 CN1424402A (en) | 2002-12-12 | 2002-12-12 | Niacinamide production by microbiological catalysis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02151290 CN1424402A (en) | 2002-12-12 | 2002-12-12 | Niacinamide production by microbiological catalysis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1424402A true CN1424402A (en) | 2003-06-18 |
Family
ID=4751976
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 02151290 Pending CN1424402A (en) | 2002-12-12 | 2002-12-12 | Niacinamide production by microbiological catalysis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1424402A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102776255A (en) * | 2012-07-27 | 2012-11-14 | 马鞍山优科化工有限公司 | Preparation method of nicotinamide |
| CN104762338A (en) * | 2015-03-17 | 2015-07-08 | 安徽国星生物化学有限公司 | Method of producing nicotinamide by catalysis of rhodococcus |
| CN104762340A (en) * | 2015-03-17 | 2015-07-08 | 安徽瑞邦生物科技有限公司 | Method of converting nicotinonitrile into nicotinamide with immobilized cells |
| CN104762339A (en) * | 2015-03-17 | 2015-07-08 | 安徽国星生物化学有限公司 | Method of synthesizing nicotinamide by catalyzing 3-cyanopyridine through induction of brevibacterium flavum by rare earth metal ions |
| CN106367447A (en) * | 2016-08-28 | 2017-02-01 | 山东昆达生物科技有限公司 | Preparation method of nicotinamide |
| CN110129387A (en) * | 2019-04-18 | 2019-08-16 | 兄弟科技股份有限公司 | The method that composite material immobilization propionic acid bar bacterium prepares niacinamide |
| CN110846355A (en) * | 2019-12-02 | 2020-02-28 | 山东昆达生物科技有限公司 | Preparation method of isonicotinamide |
-
2002
- 2002-12-12 CN CN 02151290 patent/CN1424402A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102776255A (en) * | 2012-07-27 | 2012-11-14 | 马鞍山优科化工有限公司 | Preparation method of nicotinamide |
| CN102776255B (en) * | 2012-07-27 | 2014-05-14 | 马鞍山优科化工有限公司 | Preparation method of nicotinamide |
| CN104762338A (en) * | 2015-03-17 | 2015-07-08 | 安徽国星生物化学有限公司 | Method of producing nicotinamide by catalysis of rhodococcus |
| CN104762340A (en) * | 2015-03-17 | 2015-07-08 | 安徽瑞邦生物科技有限公司 | Method of converting nicotinonitrile into nicotinamide with immobilized cells |
| CN104762339A (en) * | 2015-03-17 | 2015-07-08 | 安徽国星生物化学有限公司 | Method of synthesizing nicotinamide by catalyzing 3-cyanopyridine through induction of brevibacterium flavum by rare earth metal ions |
| CN106367447A (en) * | 2016-08-28 | 2017-02-01 | 山东昆达生物科技有限公司 | Preparation method of nicotinamide |
| CN110129387A (en) * | 2019-04-18 | 2019-08-16 | 兄弟科技股份有限公司 | The method that composite material immobilization propionic acid bar bacterium prepares niacinamide |
| CN110129387B (en) * | 2019-04-18 | 2020-08-25 | 兄弟科技股份有限公司 | Method for preparing nicotinamide by composite material immobilized corynebacterium propionate |
| CN110846355A (en) * | 2019-12-02 | 2020-02-28 | 山东昆达生物科技有限公司 | Preparation method of isonicotinamide |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111304106B (en) | Bacillus clausii and method for producing tetrahydropyrimidine by using same | |
| Häggström et al. | Calcium alginate immobilized cells of Clostridium acetobutylicum for solvent production | |
| CN108410783B (en) | Method for producing glucosamine by culturing escherichia coli and fermenting | |
| CN106635934B (en) | Thermophilic lactobacillus and corn soaking method by artificially adding thermophilic lactobacillus | |
| CN102925502A (en) | Industry method for producing arachidonic acid grease by using mortierella alpine | |
| CN1884308A (en) | Process for preparing chitosan oligosacharides | |
| CN1730660A (en) | Microorganism catalytic processes for niacinamide production | |
| CN1424402A (en) | Niacinamide production by microbiological catalysis | |
| CN113046253B (en) | Culture method for improving heat resistance of kluyveromyces marxianus | |
| CN104531810B (en) | A kind of method that high-effective microorganism conversion prepares maltobionic acid | |
| CN1699585A (en) | Process for preparing biological antibacterial agent | |
| CN1524961A (en) | Microorganism continuous catalysis method for producing acrylamide | |
| CN100390295C (en) | Microorganism polysaccharide and its preparation method and application | |
| CN114958631B (en) | Method for producing single cell protein by using heavy phase lactic acid | |
| CN114058654B (en) | Fermentation method for increasing yield of gamma-aminobutyric acid | |
| CN104762340A (en) | Method of converting nicotinonitrile into nicotinamide with immobilized cells | |
| CN101575579A (en) | Ferrum-rich saccharomyces cerevisiae and production method thereof | |
| CN111424061B (en) | Rhodococcus ruber and method for producing nicotinamide by using same | |
| CN1133746C (en) | Process for preparing xylitol by repeated use of free cells and multiple transforms | |
| CN103865901B (en) | A kind of fermention medium of saccharifying enzyme and fermentation process thereof | |
| CN1308456C (en) | Microorganism catalysis method for producing acrylamide | |
| CN1259427C (en) | Method for preparing grape manna oligosaccharide | |
| CN114317641A (en) | Optimized fermentation process of N-acetylglucosamine engineering bacteria | |
| CN114196713B (en) | Method for reducing carbon dioxide emission in glucosamine fermentation process | |
| CN102286569A (en) | Curdlan and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |