CN106086103A - A kind of separating and extracting process of feed grade nicotinic acid - Google Patents
A kind of separating and extracting process of feed grade nicotinic acid Download PDFInfo
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- CN106086103A CN106086103A CN201610542267.2A CN201610542267A CN106086103A CN 106086103 A CN106086103 A CN 106086103A CN 201610542267 A CN201610542267 A CN 201610542267A CN 106086103 A CN106086103 A CN 106086103A
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- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
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- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
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- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
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Abstract
The invention discloses and a kind of utilize Multistage Membranes partition method and the spray drying technology method of separation and Extraction feed grade nicotinic acid from bioconversion broth, including with immobilized cell as catalyst, with 3 cyanopyridines as substrate, carry out bioconversion and obtain the reactant liquor containing nicotinic acid, use the micro-filtration membrane of 100nm 10 μm, molecular cut off is the ultrafilter membrane of 1000 3000Da, the Multistage Membranes isolation technics such as the NF membrane of 200 300Da remove particulate matter in conversional solution, solubility foreign protein, the impurity such as pigment, make conversional solution concentrate simultaneously, it is dried at spray dryer, inlet temperature is 210 240 DEG C, outlet temperature is 60 100 DEG C, nicotinic acid product is i.e. obtained through being spray-dried, nicotinic acid extracts total recovery and can reach more than 85%.The method is easy and simple to handle, with low cost, product yield is high, pollutant effluents discharge capacity is few, has a good application prospect.
Description
Technical field
The invention belongs to biological chemical field, relate to one and prepare extraction, particularly relate to separate from bioconversion broth
The method extracting feed grade nicotinic acid.
Background technology
Nicotinic acid is white or off-white powder, the most also referred to as nicotinic acid, vitamin PP or vitamin B3, is pig, chicken
Deng composition necessary in animal diets, can as vitamins feed additive (Zhao Dongjiang, chemical industry in Jiangsu Province, 2005,33:53-
56).In animal body, nicotinic acid is to constitute nadide and coenzyme II important as precursors, and the Main Function of these coenzyme is that promotion is dynamic
The transfer of hydrogen in object, has synthesis and the acid that reduces fat, carbohydrate and amino acid whose basic function.Nadide and coenzyme
II is also tricarboxylic acids or the ingredient of Kreb circulation.This circulation is the metabolism of carbohydrate oxidation in animal body
Activity, energy and utilization thereof that metabolism produces have to rely on them and complete, thus play important work in animal physiological function
With (Li Yanyun, Beijing Institute of Petrochemical Technology's journal, 2006,14:59-64).In view of these functions of nicotinic acid, this compound at present
It is widely used (Jiang Guangcheng, veterinary drug feed additive, 1997,1:11-12) at feedstuff industry tool.
Up to now the production method of nicotinic acid includes chemical oxidization method and biotransformation method.Nicotinic acid preparation the earliest produces work
Before skill builds on 100 years, mainly used at that time potassium dichromate oxidation nicotine prepare nicotinic acid (Chuck R., Chimia,
2000,54:508-513).The existing industrial process of nicotinic acid includes potassium permanganate oxidation method, nitre potassium permanganate oxidation method, nitre
Nitric acid-sulfuric acid oxidizing process, ammonia oxidation etc. (Wang Liucheng etc., fine chemistry industry, 2004,21:133-136).But these existing methods
Needing to carry out under high pressure, hot conditions, corrosivity is big, and the requirement to equipment is high, and production cost is high, and economic benefit is low, the three wastes
Seriously polluted.Compared to chemical method, utilizing biological enzyme synthesis nicotinic acid to have course of reaction simple, by-product is few, technique bar
Part is gentle, and the most progressively come into one's own (Gong JS, et al.Microb Cell Fact, 2012,11:142), biological turn
Change method is current prefered method, and its product extracts flow process and generally includes: the acidifying of microbe conversion liquid, ceramic membrane filter, ultrafiltration, work
Property carbon decoloring, crystallization.The method extracting cycle length, acid adding waste water intractability are big, high to equipment investment cost, yield is the most inclined
Low.The present invention uses the bacillus subtilis immobilized cell containing nitrilase to carry out being converted into nicotinic acid, uses microfiltration and surpasses
Filter miscellaneous, remove the particulate matter in fermentation liquid, solubility foreign protein, and make fermentation liquid be concentrated, at spray dryer simultaneously
It is dried (inlet temperature is 210-240 DEG C, and outlet temperature is 60-100 DEG C), obtains nicotinic acid product, product quality and yield
Height, process costs is low.Production process, without using a large amount of concentrated acid, concise in technology, is greatly reduced discharge of wastewater, energy-conserving and environment-protective,
And can realize producing continuously.Nicotinic acid extracts total recovery and can reach more than 85%, and product quality meets national standard GB/T 7300-
2006, feed additive nicotinic acid.Wang Daming et al. once used the inlet temperature of 200 DEG C and 250 DEG C to carry out spray drying to prepare cigarette
Acid (Wang Daming, Hebei Normal University's master thesis, 2012), significantly different with this research.Immobilization is passed through in this research
Cell carries out bioconversion, has reacted rear immobilized cell and can have been filtered by a step and quickly remove, post processing without degerming body,
Saved process costs and operating procedure, existing document report use immobilization gibberella Gibberella intermedia with
And pseudomonas putida Pseudomonas putida prepares nicotinic acid for bioconversion and achieves good result (Tao Yang etc., change
Work is in progress, 2014,33:2432-2437;Lee is identical, food and biotechnology journal, and 2014,33:800-804), this research is adopted
Carry out bioconversion synthesis nicotinic acid with bacillus subtilis immobilized cell to still belong to the first time report.
Summary of the invention
The present invention uses a kind of immobilized cell to carry out bioconversion and generates nicotinic acid, uses microfiltration and ultrafiltration remove impurity, removes
Particulate matter, solubility foreign protein, and make fermentation liquid be concentrated simultaneously, it is dried at spray dryer, obtains nicotinic acid product,
Product quality and yield are high, and process costs is low.The operation of the method more original method is easier, cost is lower, also improve product
Product yield, reduce highly polluted wastewater displacement.
It is an object of the invention to provide a kind of convenient and swift, low cost, effective extraction of preparing in fermentation liquid is raised
The method of material level nicotinic acid.
Therefore it is an object of the invention to provide a kind of from fermentation liquid extraction prepare the technical solution of nicotinic acid, specifically
Comprise the steps:
A kind of immobilized cell carries out the method that bioconversion generates nicotinic acid, and described method includes: to produce nitrilase
Bacillus subtilis immobilized cell is catalyst, with nicotinonitrile as substrate, carries out biological turning under the conditions of 20-50 DEG C
Change, react 5-30h, it is thus achieved that the reactant liquor containing nicotinic acid, Nicotinic is 150-450g/L.
Described reactant liquor can remove immobilized cell by filtering, and obtains the filtrate containing nicotinic acid, and immobilized cell reclaims also
Convert for lower batch;
Described containing nicotinic acid filtrate in temperature 20-45 DEG C, pressure is to pass through metal oxide film in the case of 1.0-1.5MPa
Or ceramic membrane filter, the aperture of film is 100nm-10 μm, microfiltration to volume concentration than 2-5 times time, start filter wash, collection passes through
Liquid;
Described microfiltration permeate is 25-40 DEG C in temperature, and pressure is to pass through molecular cut off in the case of 0.05-0.1MPa
For the ultrafilter membrane of 1000-3000Da, ultrafiltration to volume concentration ratio, for during 6-8, collects permeate;
Described ultrafiltration permeate is 10-40 DEG C in temperature, and pressure is to use receiving of 200-300Da in the case of 0.1-2Mpa
Filter membrane removes the small molecular weight impurities such as depigmentation, polypeptide, nucleotide, moisture.
Described nanofiltration permeate uses membrane evaporator to be concentrated in vacuo, and concentrated solution is dried at spray dryer, enters
Mouth temperature is 210-240 DEG C, and outlet temperature is 60-100 DEG C, i.e. obtains nicotinic acid product through being spray-dried, and nicotinic acid extracts total receipts
Rate can reach more than 85%.
Advantages of the present invention and good effect be:
1, the present invention uses immobilized cell to carry out nicotinic acid synthesis as catalyst, compares traditional free cell synthesis work
Skill has the advantage that 1. immobilized cell is conducive to the sharp separation of nicotinic acid product to extract as catalyst, it is not necessary to arrange trip
Link is removed from cell;2. the reaction of immobilized cell is advantageously implemented the recycling of catalyst, can save production cost;③
Immobilization is conducive to promoting the stability of cell, can extend cell and use the cycle;4. the heavy industrialization of nicotinic acid it is easily achieved
Synthesis.
2, the present invention uses the step such as microfiltration and ultrafiltration to go the removal of impurity, first passes through the microfiltration of metal oxide film or ceramic membrane
Effect, removes macromolecular particle thing and small-sized immobilized cell and fragment etc., then is 1000-3000Da's by molecular cut off
Hollow fiber ultrafiltration membrane removes the impurity such as the protein in filtrate, inorganic salt, removes depigmentation, polypeptide, nucleotide, water by nanofiltration
Grade small molecular weight impurity, uses three step remove impurity, it is ensured that effectively removes the various impurity in solution, makes nicotinic acid product
Purity is higher, has highly important practice significance.
3, this method use spray drying obtain can direct applied nicotinic acid finished product, have that equipment is simple, low cost, be suitable for
In advantages such as large-scale industrial production, the nicotinic acid performance safety obtained, purity are high, white uniform powder.
Detailed description of the invention
Following example are only used for describing the present invention, but the present invention is not limited to these embodiments restrictions.
Embodiment 1
In the reaction system of 5L, with bacillus subtilis immobilized cell as catalyst, with nicotinonitrile as substrate,
Under the conditions of 30 DEG C, carry out bioconversion, add substrate by batch and carry out conversion reaction 7h, it is thus achieved that the reactant liquor containing nicotinic acid, cigarette
Acid concentration is 184g/L.Filter and remove immobilized cell, obtain filtrate.Filtrate containing nicotinic acid at temperature 30 DEG C, pressure is
By ceramic membrane filter in the case of 1.0MPa, the aperture of film is 5 μm, collects permeate.Permeate is 30 DEG C in temperature, pressure
For being the ultrafilter membrane of 1000Da by molecular cut off in the case of 0.1MPa, collecting filtrate, be 30 DEG C in temperature, pressure is
The NF membrane using 300Da in the case of 0.1MPa removes small molecular weight impurity.Permeate uses membrane evaporator to be concentrated in vacuo, by dense
Contracting liquid is dried at spray dryer, and inlet temperature is 210 DEG C, and outlet temperature is 80 DEG C, i.e. obtains cigarette through being spray-dried
Acid product, products obtained therefrom is white powder, and nicotinic acid extracts total recovery and can reach 86.9%.
Embodiment 2
In the reaction system of 10L, with bacillus subtilis immobilized cell as catalyst, with nicotinonitrile as the end
Thing, carries out bioconversion under the conditions of 30 DEG C, adds substrate by batch and carries out conversion reaction 10h, it is thus achieved that the reaction containing nicotinic acid
Liquid, Nicotinic is 246g/L.Filter and remove immobilized cell, obtain filtrate.Filtrate containing nicotinic acid at temperature 30 DEG C, pressure is
By ceramic membrane filter in the case of 1.2MPa, the aperture of film is 1.0 μm, starts filter wash, collects permeate.Permeate is in temperature
Being 30 DEG C, pressure is in the case of 0.1MPa to be the ultrafilter membrane of 1000Da by molecular cut off, collects filtrate, is 30 in temperature
DEG C, pressure is that the NF membrane using 200Da in the case of 0.1MPa removes small molecular weight impurity.Permeate uses membrane evaporator vacuum
Concentrating, be dried at spray dryer by concentrated solution, inlet temperature is 220 DEG C, and outlet temperature is 60 DEG C, through being spray-dried
I.e. obtaining nicotinic acid product, nicotinic acid extracts total recovery and can reach 89.4%.
Embodiment 3
In the reaction system of 5L, with bacillus subtilis immobilized cell as catalyst, with nicotinonitrile as substrate,
Under the conditions of 35 DEG C, carry out bioconversion, convert by the way of multiple batches of current adding substrate.Every batch reaction 4h, it is thus achieved that cigarette
Acid concentration is 123g/L, and batch is collected by filtration immobilized cell after having reacted, and is placed in new 5L reaction system, carries on
Bottoms stream adds conversion, obtains 123g/L nicotinic acid by 4h reaction;So circulation 5 batches, through the 16h response time, finally accumulate
25L nicotinic acid solution, concentration is 123g/L.Filtrate containing nicotinic acid is temperature 30 DEG C, and pressure is by pottery in the case of 1.5MPa
Membrane filtration, the aperture of film is 1.0 μm, collects permeate.Permeate is 30 DEG C in temperature, and pressure is by cutting in the case of 0.1MPa
The ultrafilter membrane staying molecular weight to be 1000Da, collects filtrate, is 30 DEG C in temperature, and pressure is employing 200Da in the case of 0.2MPa
NF membrane removes small molecular weight impurity.Permeate uses membrane evaporator to be concentrated in vacuo, and is done at spray dryer by concentrated solution
Dry, inlet temperature is 210 DEG C, and outlet temperature is 80 DEG C, i.e. obtains nicotinic acid product through being spray-dried, and nicotinic acid extracts total recovery can
Reach 91.2%.
The invention is not restricted to these disclosed embodiments, the present invention is by the scope described by soverlay technique scheme, Yi Jiquan
The various deformation of profit claimed range and equivalence change, on the premise of without departing from the technical solution of the present invention, to the present invention
Any amendment or improvement that the those skilled in the art made easily realize belong to scope of the present invention.
Claims (7)
1. a method for separation and Extraction feed grade nicotinic acid from bioconversion broth, is characterized by, fix with bacillus subtilis
Change cell is catalyst, carries out the bioconversion acquisition reactant liquor containing nicotinic acid with nicotinonitrile for substrate, uses microfiltration and surpasses
The multi-stage membrane separation process such as filter remove the impurity such as particulate matter, solubility foreign protein, pigment, make conversional solution be concentrated simultaneously,
Spray dryer is dried, obtains nicotinic acid product.
2. the method for claim 1, is characterized by, described reactant liquor contains nicotinic acid, to produce the withered of nitrilase
Grass bacillus cereus immobilized cell is catalyst, with nicotinonitrile as substrate, carries out bioconversion under the conditions of 20-50 DEG C,
Reaction 5-30h, prepares.
3. the method for claim 1, is characterized by, described reactant liquor can be removed by filtration and reclaim fixation cell
Born of the same parents, the immobilized cell of recovery can be used for lower batch and converts, filters containing nicotinic acid in gained filtrate, and Nicotinic is 150-
450g/L。
4. the method for claim 1, is characterized by, the multi-stage membrane separation process such as described employing microfiltration and ultrafiltration, including
By reactant liquor in temperature 20-45 DEG C, pressure is by metal oxide film or ceramic membrane filter in the case of 1.0-1.5MPa, film
Aperture is 100nm-10 μm, collects permeate.
5. the method for claim 1, is characterized by, the concrete operations of described microfiltration are, is 25-by reactant liquor in temperature
40 DEG C, pressure is in the case of 0.05-0.1MPa to be the ultrafilter membrane of 1000-3000Da by molecular cut off, collects filtrate.
6. the method for claim 1, is characterized by, the concrete operations of described ultrafiltration are, is 10-by reactant liquor in temperature
40 DEG C, pressure is to use the NF membrane of 200-300Da to remove little point of depigmentation, polypeptide, nucleotide, moisture etc. in the case of 0.1-2MPa
Sub-impurity.
7. the method for claim 1, is characterized by, the concrete operations of described spray drying are, permeate concentrated
Spray dryer is dried, and inlet temperature is 210-240 DEG C, and outlet temperature is 60-100 DEG C, i.e. obtains through being spray-dried
Nicotinic acid product, nicotinic acid extracts total recovery and can reach more than 85%.
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Cited By (6)
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CN109055454A (en) * | 2018-10-24 | 2018-12-21 | 张刘兵 | A kind of separating and extracting process of feed grade niacin |
CN109055453A (en) * | 2018-09-19 | 2018-12-21 | 安徽瑞邦生物科技有限公司 | A kind of separating and extracting process of feed grade niacin |
CN109136301A (en) * | 2018-09-19 | 2019-01-04 | 安徽瑞邦生物科技有限公司 | A kind of method that hydrolysis prepares niacin |
CN110746351A (en) * | 2019-10-30 | 2020-02-04 | 南宁学院 | Method for extracting nicotinic acid from beef tripe |
CN111778199A (en) * | 2020-09-07 | 2020-10-16 | 鲁东大学 | Bacillus subtilis and application thereof |
CN114456107A (en) * | 2020-11-10 | 2022-05-10 | 安徽瑞邦生物科技有限公司 | Method for producing isonicotinic acid by using isonicotinic acid ammonium as raw material |
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Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109055453A (en) * | 2018-09-19 | 2018-12-21 | 安徽瑞邦生物科技有限公司 | A kind of separating and extracting process of feed grade niacin |
CN109136301A (en) * | 2018-09-19 | 2019-01-04 | 安徽瑞邦生物科技有限公司 | A kind of method that hydrolysis prepares niacin |
CN109055454A (en) * | 2018-10-24 | 2018-12-21 | 张刘兵 | A kind of separating and extracting process of feed grade niacin |
CN110746351A (en) * | 2019-10-30 | 2020-02-04 | 南宁学院 | Method for extracting nicotinic acid from beef tripe |
CN110746351B (en) * | 2019-10-30 | 2021-03-16 | 南宁学院 | Method for extracting nicotinic acid from beef tripe |
CN111778199A (en) * | 2020-09-07 | 2020-10-16 | 鲁东大学 | Bacillus subtilis and application thereof |
CN111778199B (en) * | 2020-09-07 | 2020-11-24 | 鲁东大学 | Bacillus subtilis and application thereof |
CN114456107A (en) * | 2020-11-10 | 2022-05-10 | 安徽瑞邦生物科技有限公司 | Method for producing isonicotinic acid by using isonicotinic acid ammonium as raw material |
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