CN104293851A - Method for producing hydroxyethyl pyridine by alternaria alternata - Google Patents
Method for producing hydroxyethyl pyridine by alternaria alternata Download PDFInfo
- Publication number
- CN104293851A CN104293851A CN201410594938.0A CN201410594938A CN104293851A CN 104293851 A CN104293851 A CN 104293851A CN 201410594938 A CN201410594938 A CN 201410594938A CN 104293851 A CN104293851 A CN 104293851A
- Authority
- CN
- China
- Prior art keywords
- diatomite
- substrate
- reaction
- pyridine
- ethyl acetate
- 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
- BXGYBSJAZFGIPX-UHFFFAOYSA-N 2-pyridin-2-ylethanol Chemical compound OCCC1=CC=CC=N1 BXGYBSJAZFGIPX-UHFFFAOYSA-N 0.000 title abstract description 3
- 241000223602 Alternaria alternata Species 0.000 title abstract 4
- 238000004519 manufacturing process Methods 0.000 title abstract 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 58
- 239000000758 substrate Substances 0.000 claims abstract description 57
- 238000006243 chemical reaction Methods 0.000 claims abstract description 44
- 238000000034 method Methods 0.000 claims abstract description 23
- 239000000872 buffer Substances 0.000 claims abstract description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 36
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 28
- 241000894006 Bacteria Species 0.000 claims description 23
- 239000000243 solution Substances 0.000 claims description 16
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 14
- 230000002210 biocatalytic effect Effects 0.000 claims description 12
- 238000009423 ventilation Methods 0.000 claims description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 240000008042 Zea mays Species 0.000 claims description 7
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 7
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 7
- 208000012839 conversion disease Diseases 0.000 claims description 7
- 235000005822 corn Nutrition 0.000 claims description 7
- 229960003487 xylose Drugs 0.000 claims description 7
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 claims description 6
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims description 6
- 238000003810 ethyl acetate extraction Methods 0.000 claims description 6
- 238000000605 extraction Methods 0.000 claims description 6
- 239000002953 phosphate buffered saline Substances 0.000 claims description 6
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 6
- 229920000053 polysorbate 80 Polymers 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 4
- XVMSFILGAMDHEY-UHFFFAOYSA-N 6-(4-aminophenyl)sulfonylpyridin-3-amine Chemical compound C1=CC(N)=CC=C1S(=O)(=O)C1=CC=C(N)C=N1 XVMSFILGAMDHEY-UHFFFAOYSA-N 0.000 abstract 1
- 150000003016 phosphoric acids Chemical class 0.000 abstract 1
- 238000006555 catalytic reaction Methods 0.000 description 8
- 230000008569 process Effects 0.000 description 6
- BEZVGIHGZPLGBL-UHFFFAOYSA-N 2,6-diacetylpyridine Chemical compound CC(=O)C1=CC=CC(C(C)=O)=N1 BEZVGIHGZPLGBL-UHFFFAOYSA-N 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000004321 preservation Methods 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 240000008564 Boehmeria nivea Species 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000005515 coenzyme Substances 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- 101000925662 Enterobacteria phage PRD1 Endolysin Proteins 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003905 agrochemical Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000012847 fine chemical Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000024287 Areas Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 241000790917 Dioxys <bee> Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 101710157860 Oxydoreductase Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 238000011914 asymmetric synthesis Methods 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 230000005389 magnetism Effects 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a method for producing hydroxyethyl pyridine by alternaria alternata. Accoring to the method, alternaria alternata cells are utilized to prepare the (S, S)-2,6-(1-hydroxyethyl) pyridine in a biocatalysis manner, a phosphoric acid salt buffer is added into a reaction tank, and diatomite is added into to adjust the concentrations of both a substrate and a product. According to the method, with the alternaria alternata cells, the biocatalysis reaction is carried out, the yield of the product is high, the enantiomeric excess rate (ee%) is high, and an excellent application prospect is provided.
Description
Technical field
The invention belongs to technology of pharmaceutical engineering field, be related specifically to alternaric bacteria cell biocatalysis and prepare chiral medicinal intermediate (S, S)-2,6-(1-hydroxyethyl) technology of pyridine.
Background technology
Chirality (S, S)-2,6-(1-hydroxyethyl) pyridine is the important intermediate of synthesis of chiral medicine, fine chemicals, agricultural chemicals product and other exotic materialss.Biocatalysis asymmetric reaction has environmental friendliness, mild condition, selectivity advantages of higher, prepares the first approach of hydroxyl chipal compounds as green high-efficient, is applied to the chipal compounds producing some high added values more and more.Biocatalysis prepares chirality (S, S)-2,6-(1-hydroxyethyl) pyridine has good application prospect.
Biocatalysis has the outstanding advantages such as catalytic efficiency is high, selectivity strong, mild condition, environmental friendliness, is the important method substituting and expand traditional organic chemical synthesis in process of sustainable development.Wherein chiral separation and asymmetric synthesis are the Application Areass of biocatalysis most magnetism.As in six large fermentoids of biological catalyst, lytic enzyme catalytic kinetics resolution of racemates can obtain quiral products, in industrial biocatalytic, play key player always.In recent years, oxydo-reductase application industrially obtained and increased rapidly.At present, the ratio adopting the industry of lytic enzyme Kinetic Resolution, biological catalysis and biological oxidation process to prepare optical activity chirality compound is 4:2:1.Biomass cells reduction method is for Kinetic Resolution, and maximum advantage is that theoretical yield can reach 100%, and Atom economy is good.But bioreduction needs the participation of coenzyme or cofactor, limit its application to a certain extent.Due to the dependent cause of coenzyme, in bioreduction, many Bian intact cells are as catalyzer, realize the purification procedures eliminating enzyme in body while coenzyme cyclic regeneration.
(S, S)-2,6-(1-hydroxyethyl) pyridine is the important chiral building block of synthesis of chiral medicine, fine chemicals, agricultural chemicals product, owing to there being 2 chiral centres, make chemosynthesis comparatively difficult, cost is high.The present invention will adopt biocatalysis preparation (S, S)-2,6-(1-hydroxyethyl) pyridine.
Summary of the invention
The present invention adopts alternaric bacteria cell catalysis preparation (S, S)-2,6-(1-hydroxyethyl) pyridine, reaction formula is as follows:
Substrate DAP (1), through alternaric bacteria catalyzed reaction, obtains product (S, S)-2,6-(1-hydroxyethyl) pyridine (2).Have multiple-microorganism can catalysis this reaction, through great many of experiments screening, finally determine adopt alternaric bacteria as catalyzer because its catalysis 1 reaction effect best, reaction yield, enantiomeric excess rate (ee%) are all very high.
Many alternaric bacterias can carry out this reaction of biocatalysis, but its effect is different, differs greatly, and through experiment, the present invention selects alternaric bacteria bacterial strain to be ATCC 66983, and its this reaction effect of catalysis is best.
developing medium:
1, nutrient solution composition: corn steep liquor (with dry basis) content is 35-45g/L; analysis for soybean powder 3-5g/L; yeast extract 6-9g/L, glucose 18-22 g/L, malt extract 25-30 g; ammonium sulfate 0.5-0.7g/L; magnesium sulfate 0.3-0.4g/L, phosphoric acid dioxy potassium 1.8-2.3g/L, calcium carbonate 0.7 g/L; ferrous sulfate 0.18 g/L, manganous sulfate 0.025 g/L; PH 5.3.
2, solid medium composition: the agar powder adding 1.5-2% in liquid medium within.
Prepared by alternaric bacteria wet cell.Alternaric bacteria through inclined-plane, shaking flask, seed tank culture obtain seed liquor; Fermentor tank adds nutrient solution, and coefficient is 0.6-0.7, and 121 DEG C of autoclavings 30 minutes, are cooled to 28-29 DEG C, by alternaric bacteria
aTCC 66983seed liquor is seeded to fermentor tank, and inoculative proportion is 10-15%, and ventilation ratio is 0.5-1V/(V minute), namely per minute air flow is 0.5-1 times of fermentating liquid volume, cultivate 36-40 hour, obtain wet alternaric bacteria cell, as biocatalytic reaction catalyzer with filtering centrifuge is centrifugal for 28-29 DEG C.Wet cell preparation technique is mature technology.
Because substrate, product all have restraining effect to alternaric bacteria cell, in order to reduce the suppression of substrate, products upon cell, the present invention adopts diatomite adsorption substrate, in reaction, when substrate is reacted by cell catalysis, when concentration reduces, substrate from diatomite stripping, postreaction consume substrate.Meanwhile, product, by diatomite adsorption, decreases its concentration in water, thus considerably reduces the suppression of substrate, products upon cell.Substrate and diatomaceous ratio, determine the concentration of substrate in reaction solution, product, and concentration is also relevant with temperature of reaction, reaction solution composition.Different cell is different to the susceptibility of substrate, product, so, great many of experiments be carried out, best substrate and diatomaceous ratio could be determined.Experiment shows, for bacterial strain of the present invention, reaction solution composition, temperature of reaction, best substrate and diatomaceous ratio are 0.25-0.28.During concrete absorption, substrate DAP diatomite is absorbed, namely obtains the diatomite having adsorbed substrate.Regulate substrate and diatomite consumption, make substrate and diatomaceous quality ratio be 0.25-0.28.Ramie gauze used is common ramie gauze, and commercially, sterilizing, Preservation in sterile condition is stand-by.
Phosphate buffered saline buffer is added in bottom ventilation stirred tank, pH is 6.4, add the diatomite having adsorbed substrate DAP, make substrate 2,6-diacetyl pyridine addition is 80-90 g/L, corn steep liquor (with dry basis) content is 20-22g/L, and glucose content is 5-7g/L, wood sugar 7-9 g/L, tween 80 content is 16-19g/L, 121 DEG C of autoclavings 30 minutes; When being cooled to 31-32 DEG C, add wet alternaric bacteria cell and make concentration be 33-36g/L, ventilation ratio is 0.16-0.19V/(V minute), namely per minute air flow is 0.16-0.19 times of reaction solution volume, carries out biocatalytic reaction, and the reaction times is 37-42 hour; After reaction terminates, leach cell, diatomite respectively, be extracted with ethyl acetate reaction solution, extraction diatomite, combined ethyl acetate extraction liquid, steams ethyl acetate, obtain product (S, S)-2,6-(1-hydroxyethyls) pyridine, reaction conversion ratio 96-98%, product yield 94-96%, enantiomeric excess rate (ee%) 98-99%.
The present invention carries out the work and comprises bacterial strain selection (selecting from more or less a hundred bacterial strain), catalytic reaction condition optimizes (temperature of reaction, air flow, pH), reaction medium is selected and concentration optimization (concentration of substrate, glucose content, Xylose Content, kinds of surfactants (more than 20, kind selects 1) and concentration, other multiple components is selected to get rid of), solid absorption is adopted to control substrate product concentration, thus the suppression reduced cell, test carclazyte, diatomite, cotton gauze, ramie gauze, the many kinds of solids materials such as macropore resin, also once carried out the test of water-organic solvent 2 phase system.Because tested number is very large, although have employed response surface optimization design experiment, drastically reduce the area tested number, tested number is still very large, and total Test carries out just completing more than 2 years, reaches current technical scheme.For photolytic activity product, as reaction conversion ratio 96-98%, during product yield 94-96%, enantiomeric excess rate (ee%), still up to 98-99%, is very not easily, and our work achieves marked improvement.
embodiment 1
Wet alternaric bacteria ATCC 66983 cell is produced, as biocatalytic reaction catalyzer by ordinary method.
The diatomite making method of having adsorbed substrate is, is absorbed by substrate DAP diatomite, namely obtains the diatomite having adsorbed substrate, regulates substrate and diatomite consumption, makes substrate and diatomaceous quality ratio be 0.25.Diatomite used is common diatomite, and commercially, sterilizing, Preservation in sterile condition is stand-by, and other embodiment diatomite process is identical.
Phosphate buffered saline buffer is added in 15L bottom ventilation stirred tank, pH is 6.4, add the diatomite having adsorbed substrate DAP, make substrate 2,6-diacetyl pyridine addition is 80 g/L, corn steep liquor (with dry basis) content is 20g/L, and glucose content is 5g/L, wood sugar 7 g/L, tween 80 content is 16g/L, 121 DEG C of autoclavings 30 minutes; When being cooled to 31 DEG C, add wet alternaric bacteria cell and make concentration be 33g/L, ventilation ratio is 0.16V/(V minute), namely per minute air flow is 0.16 times of reaction solution volume, carries out biocatalytic reaction, and the reaction times is 37 hours; After reaction terminates, leach cell, diatomite respectively, be extracted with ethyl acetate reaction solution, extraction diatomite, combined ethyl acetate extraction liquid, steams ethyl acetate, obtain product (S, S)-2,6-(1-hydroxyethyls) pyridine, reaction conversion ratio 96%, product yield 94%, enantiomeric excess rate (ee%) 99%.
embodiment 2
Wet alternaric bacteria ATCC 66983 cell is produced, as biocatalytic reaction catalyzer by ordinary method.
The diatomite making method of having adsorbed substrate is, is absorbed by substrate DAP diatomite, namely obtains the diatomite having adsorbed substrate, regulates substrate and diatomite consumption, makes substrate and diatomaceous quality ratio be 0.28.Diatomite used is common diatomite, and commercially, sterilizing, Preservation in sterile condition is stand-by, and other embodiment diatomite process is identical.
Phosphate buffered saline buffer is added in 15L bottom ventilation stirred tank, pH is 6.4, add the diatomite having adsorbed substrate DAP, make substrate 2,6-diacetyl pyridine addition is 90 g/L, corn steep liquor (with dry basis) content is 22g/L, and glucose content is 7g/L, wood sugar 9 g/L, tween 80 content is 19g/L, 121 DEG C of autoclavings 30 minutes; When being cooled to 32 DEG C, add wet alternaric bacteria cell and make concentration be 36g/L, ventilation ratio is 0.19V/(V minute), namely per minute air flow is 0.19 times of reaction solution volume, carries out biocatalytic reaction, and the reaction times is 42 hours; After reaction terminates, leach cell, diatomite respectively, be extracted with ethyl acetate reaction solution, extraction diatomite, combined ethyl acetate extraction liquid, steams ethyl acetate, obtain product (S, S)-2,6-(1-hydroxyethyls) pyridine, reaction conversion ratio 98%, product yield 96%, enantiomeric excess rate (ee%) 98%.
embodiment 3
Wet alternaric bacteria ATCC 66983 cell is produced, as biocatalytic reaction catalyzer by ordinary method.
The diatomite making method of having adsorbed substrate is, is absorbed by substrate DAP diatomite, namely obtains the diatomite having adsorbed substrate, regulates substrate and diatomite consumption, makes substrate and diatomaceous quality ratio be 0.27.Diatomite used is common diatomite, and commercially, sterilizing, Preservation in sterile condition is stand-by, and other embodiment diatomite process is identical.
Phosphate buffered saline buffer is added in 15L bottom ventilation stirred tank, pH is 6.4, add the diatomite having adsorbed substrate DAP, make substrate 2,6-diacetyl pyridine addition is 85 g/L, corn steep liquor (with dry basis) content is 21g/L, and glucose content is 6g/L, wood sugar 8 g/L, tween 80 content is 17g/L, 121 DEG C of autoclavings 30 minutes; When being cooled to 31.5 DEG C, add wet alternaric bacteria cell and make concentration be 35g/L, ventilation ratio is 0.18V/(V minute), namely per minute air flow is 0.18 times of reaction solution volume, carries out biocatalytic reaction, and the reaction times is 41 hours; After reaction terminates, leach cell, diatomite respectively, be extracted with ethyl acetate reaction solution, extraction diatomite, combined ethyl acetate extraction liquid, steams ethyl acetate, obtain product (S, S)-2,6-(1-hydroxyethyls) pyridine, reaction conversion ratio 97%, product yield 95%, enantiomeric excess rate (ee%) 98.5%.
embodiment 4
Wet alternaric bacteria ATCC 66983 cell is produced, as biocatalytic reaction catalyzer by ordinary method.
The diatomite making method of having adsorbed substrate is, is absorbed by substrate DAP diatomite, namely obtains the diatomite having adsorbed substrate, regulates substrate and diatomite consumption, makes substrate and diatomaceous quality ratio be 0.28.Diatomite used is common diatomite, and commercially, sterilizing, Preservation in sterile condition is stand-by, and other embodiment diatomite process is identical.
Phosphate buffered saline buffer is added in 15L bottom ventilation stirred tank, pH is 6.4, add the diatomite having adsorbed substrate DAP, make substrate 2,6-diacetyl pyridine addition is 89 g/L, corn steep liquor (with dry basis) content is 22g/L, and glucose content is 6g/L, wood sugar 8.5g/L, tween 80 content is 18g/L, 121 DEG C of autoclavings 30 minutes; When being cooled to 31.5 DEG C, add wet alternaric bacteria cell and make concentration be 35g/L, ventilation ratio is 0.18V/(V minute), namely per minute air flow is 0.18 times of reaction solution volume, carries out biocatalytic reaction, and the reaction times is 41 hours; After reaction terminates, leach cell, diatomite respectively, be extracted with ethyl acetate reaction solution, extraction diatomite, combined ethyl acetate extraction liquid, steams ethyl acetate, obtain product (S, S)-2,6-(1-hydroxyethyls) pyridine, reaction conversion ratio 97.5%, product yield 95%, enantiomeric excess rate (ee%) 99%.
Claims (2)
1. an alternaric bacteria cell biocatalysis preparation (S, S)-2,6-(1-hydroxyethyl) method of pyridine, it is characterized in that adding phosphate buffered saline buffer in retort, pH is 6.4, add and adsorbed substrate 2, the diatomite of 6-diacetyl pyridine, make substrate DAP addition be 80-90 g/L, corn steep liquor (with dry basis) content is 20-22g/L, glucose content is 5-7g/L, wood sugar 7-9 g/L, tween 80 content is 16-19g/L, 121 DEG C of autoclavings 30 minutes; When being cooled to 31-32 DEG C, add wet alternaric bacteria cell and make concentration be 33-36g/L, ventilation ratio is 0.16-0.19V/(V minute), carry out biocatalytic reaction, the reaction times is 37-42 hour; After reaction terminates, leach cell, diatomite respectively, be extracted with ethyl acetate reaction solution, extraction diatomite, combined ethyl acetate extraction liquid, steams ethyl acetate, obtain product (S, S)-2,6-(1-hydroxyethyls) pyridine, reaction conversion ratio 96-98%, product yield 94-96%, enantiomeric excess rate (ee%) 98-99%.
2. method according to claim 1; it is characterized in that described diatomite making method of having adsorbed substrate is; by substrate 2; 6-diacetyl pyridine diatomite absorbs; namely the diatomite having adsorbed substrate is obtained; regulate substrate and diatomite consumption, make substrate and diatomaceous quality ratio be 0.25-28.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410594938.0A CN104293851A (en) | 2014-10-30 | 2014-10-30 | Method for producing hydroxyethyl pyridine by alternaria alternata |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410594938.0A CN104293851A (en) | 2014-10-30 | 2014-10-30 | Method for producing hydroxyethyl pyridine by alternaria alternata |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104293851A true CN104293851A (en) | 2015-01-21 |
Family
ID=52313799
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410594938.0A Pending CN104293851A (en) | 2014-10-30 | 2014-10-30 | Method for producing hydroxyethyl pyridine by alternaria alternata |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104293851A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114907990A (en) * | 2022-05-02 | 2022-08-16 | 中国海洋大学 | Alternaria alternata and application thereof in removing protein in waste diatomite |
| CN115029250A (en) * | 2022-05-27 | 2022-09-09 | 中国海洋大学 | Biological regeneration method of diatomite and application of diatomite in beer filtration process |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997013763A1 (en) * | 1995-10-13 | 1997-04-17 | The Penn State Research Foundation | Asymmetric synthesis catalyzed by transition metal complexes with new chiral ligands |
-
2014
- 2014-10-30 CN CN201410594938.0A patent/CN104293851A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997013763A1 (en) * | 1995-10-13 | 1997-04-17 | The Penn State Research Foundation | Asymmetric synthesis catalyzed by transition metal complexes with new chiral ligands |
Non-Patent Citations (4)
| Title |
|---|
| BAILEY D.等: "Preparation of Highly Enantiopure Pyridylethanols by Baker’s Yeast Reductions.", 《TETRAHEDRON:ASYMMETRY》 * |
| MOHAN P.等: "Bioreduction of methyl heteroaryl and aryl heteroaryl ketones in high enantiomeric excess with newly isolated fungal strains", 《BIORESOURCE TECHNOLOGY》 * |
| UNIPROT: "Q75ZG2-Q75ZG2_ALTAL", 《UNIPROT》 * |
| 曾嵘等: "生物催化羰基不对称还原合成手性醇的研究及应用进展", 《化工进展》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114907990A (en) * | 2022-05-02 | 2022-08-16 | 中国海洋大学 | Alternaria alternata and application thereof in removing protein in waste diatomite |
| CN114907990B (en) * | 2022-05-02 | 2023-07-14 | 中国海洋大学 | Alternaria alternata and application thereof in removal of protein in waste diatomite |
| CN115029250A (en) * | 2022-05-27 | 2022-09-09 | 中国海洋大学 | Biological regeneration method of diatomite and application of diatomite in beer filtration process |
| CN115029250B (en) * | 2022-05-27 | 2023-07-28 | 中国海洋大学 | Biological regeneration method of diatomite and application of diatomite in beer filtration process |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104293851A (en) | Method for producing hydroxyethyl pyridine by alternaria alternata | |
| CN104342464A (en) | Method for producing chiral phenyl methanol employing catalysis of tarlaromyces flavus | |
| CN104313074A (en) | Method for producing pyridylethanol through penicillium catalysis | |
| CN104263776A (en) | Method for producing chiral pyridine ethanol through biological catalysis | |
| CN104313075A (en) | Cell production method for photoactive bromopyridyl ethanol | |
| CN104313076A (en) | Method for producing methylpyridinyl ethanol through biological catalysis | |
| CN104263775A (en) | Method for producing 4-pyridineethanol by black mould through catalysis | |
| CN104313078A (en) | Method for producing chiral thiazolyl ethanol through biological catalysis | |
| CN104293852A (en) | Method for producing isoquinoline methyl alcohol through cell catalysis | |
| CN104357504A (en) | Cell catalysis for producing chirality methoxyl pyridine ethanol | |
| CN104388490A (en) | Method for producing chiral pyrazinyl ethanol by biological catalysis | |
| CN104313077A (en) | Biological method for producing isoxazolyl ethanol | |
| CN104313062A (en) | Method for producing photoactive phenylethanol through cell catalysis | |
| CN104293856A (en) | Method for producing fluoropyridine ethanone through cell catalysis | |
| CN104328154A (en) | Method for producing (S)-phenyl methanol from blue mold | |
| CN104212843A (en) | Method of reduction production of bromine phenyl propionic acid methyl ester through brewing yeast | |
| CN104328151A (en) | Method for producing benzothiophene ethanol by using cells as catalyst | |
| CN104328147A (en) | Production method of chlorine-contaning (2R,3S) methyl methylpropionate | |
| CN104263769A (en) | Method of producing chlorphenyl methyl propionate by biological process | |
| CN105132471A (en) | Method for producing photo-active chloro-benzene chloroethanol by virtue of microorganisms | |
| CN104263774A (en) | Method for catalytic production of chiral cyclohexylidene enol with saccharomyces cerevisiae | |
| CN105177060A (en) | Method for cell production of (S)-tetrahydronaphthol | |
| CN105039449A (en) | Method for producing (S)-furan ethanol with penicillium | |
| CN104263768A (en) | Method for producing (2R, 3S) fluorine-containing methyl propionate by using candida | |
| CN104313066A (en) | Biological reduction method for producing (2R, 3S) iodine-containing isobutyric acid-methyl ester |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150121 |
|
| RJ01 | Rejection of invention patent application after publication |