CN1696298A - Method for promoting microbe to synthesize, 1,3-propylene glycol by adding fumaric acid from extraneous sources - Google Patents
Method for promoting microbe to synthesize, 1,3-propylene glycol by adding fumaric acid from extraneous sources Download PDFInfo
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- CN1696298A CN1696298A CN 200510011917 CN200510011917A CN1696298A CN 1696298 A CN1696298 A CN 1696298A CN 200510011917 CN200510011917 CN 200510011917 CN 200510011917 A CN200510011917 A CN 200510011917A CN 1696298 A CN1696298 A CN 1696298A
- Authority
- CN
- China
- Prior art keywords
- ammediol
- fumaric acid
- synthesize
- tech grade
- glycerine
- Prior art date
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- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 title claims abstract description 44
- 239000001530 fumaric acid Substances 0.000 title claims abstract description 22
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims description 17
- YPFDHNVEDLHUCE-UHFFFAOYSA-N propane-1,3-diol Chemical compound OCCCO YPFDHNVEDLHUCE-UHFFFAOYSA-N 0.000 title abstract description 5
- 230000001737 promoting effect Effects 0.000 title 1
- 230000001580 bacterial effect Effects 0.000 claims abstract description 7
- 238000011218 seed culture Methods 0.000 claims abstract description 7
- UXFQFBNBSPQBJW-UHFFFAOYSA-N 2-amino-2-methylpropane-1,3-diol Chemical compound OCC(N)(C)CO UXFQFBNBSPQBJW-UHFFFAOYSA-N 0.000 claims description 56
- 230000004151 fermentation Effects 0.000 claims description 17
- 238000000855 fermentation Methods 0.000 claims description 15
- 244000005700 microbiome Species 0.000 claims description 6
- 238000010564 aerobic fermentation Methods 0.000 claims description 3
- 238000011177 media preparation Methods 0.000 claims 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 abstract description 47
- 235000011187 glycerol Nutrition 0.000 abstract description 21
- 238000004519 manufacturing process Methods 0.000 abstract description 17
- 239000001963 growth medium Substances 0.000 abstract description 3
- DNIAPMSPPWPWGF-VKHMYHEASA-N (+)-propylene glycol Chemical compound C[C@H](O)CO DNIAPMSPPWPWGF-VKHMYHEASA-N 0.000 abstract 1
- 229920000166 polytrimethylene carbonate Polymers 0.000 abstract 1
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 9
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 229920002215 polytrimethylene terephthalate Polymers 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- BAWFJGJZGIEFAR-NNYOXOHSSA-O NAD(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-O 0.000 description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 101000892220 Geobacillus thermodenitrificans (strain NG80-2) Long-chain-alcohol dehydrogenase 1 Proteins 0.000 description 4
- 108010025885 Glycerol dehydratase Proteins 0.000 description 4
- 230000004913 activation Effects 0.000 description 4
- RXKJFZQQPQGTFL-UHFFFAOYSA-N dihydroxyacetone Chemical compound OCC(=O)CO RXKJFZQQPQGTFL-UHFFFAOYSA-N 0.000 description 4
- 229920000139 polyethylene terephthalate Polymers 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 101710157860 Oxydoreductase Proteins 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 238000012262 fermentative production Methods 0.000 description 3
- 239000004310 lactic acid Substances 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- AKXKFZDCRYJKTF-UHFFFAOYSA-N 3-Hydroxypropionaldehyde Chemical compound OCCC=O AKXKFZDCRYJKTF-UHFFFAOYSA-N 0.000 description 2
- HGINCPLSRVDWNT-UHFFFAOYSA-N Acrolein Chemical compound C=CC=O HGINCPLSRVDWNT-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- 241000588919 Citrobacter freundii Species 0.000 description 2
- 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 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241000588747 Klebsiella pneumoniae Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- KKEYFWRCBNTPAC-UHFFFAOYSA-N Terephthalic acid Chemical compound OC(=O)C1=CC=C(C(O)=O)C=C1 KKEYFWRCBNTPAC-UHFFFAOYSA-N 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- OWBTYPJTUOEWEK-UHFFFAOYSA-N butane-2,3-diol Chemical compound CC(O)C(C)O OWBTYPJTUOEWEK-UHFFFAOYSA-N 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- -1 polytrimethylene terephthalate Polymers 0.000 description 2
- 238000007639 printing Methods 0.000 description 2
- 239000001384 succinic acid Substances 0.000 description 2
- 239000011573 trace mineral Substances 0.000 description 2
- 235000013619 trace mineral Nutrition 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- ZKHQWZAMYRWXGA-KQYNXXCUSA-N Adenosine triphosphate Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- DSCYGFBRPAPIHM-UHFFFAOYSA-N C(C)(=O)O.OCC(O)CO.C(C(O)C)(=O)O.C(CCC(=O)O)(=O)O Chemical compound C(C)(=O)O.OCC(O)CO.C(C(O)C)(=O)O.C(CCC(=O)O)(=O)O DSCYGFBRPAPIHM-UHFFFAOYSA-N 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 241000193171 Clostridium butyricum Species 0.000 description 1
- 229910021591 Copper(I) chloride Inorganic materials 0.000 description 1
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- DSQNMNVLWGRUSL-UHFFFAOYSA-N P(=O)(=O)C(C(C)=O)(O)O Chemical compound P(=O)(=O)C(C(C)=O)(O)O DSQNMNVLWGRUSL-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- LDOAKSLDUTZVSN-UHFFFAOYSA-N butane-2,3-diol;ethanol Chemical compound CCO.CC(O)C(C)O LDOAKSLDUTZVSN-UHFFFAOYSA-N 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- OXBLHERUFWYNTN-UHFFFAOYSA-M copper(I) chloride Chemical compound [Cu]Cl OXBLHERUFWYNTN-UHFFFAOYSA-M 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 238000006356 dehydrogenation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000007323 disproportionation reaction Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 229920006351 engineering plastic Polymers 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000034659 glycolysis Effects 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000013067 intermediate product Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000012092 media component Substances 0.000 description 1
- 230000007269 microbial metabolism Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000004745 nonwoven fabric Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001778 nylon Polymers 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
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- QQONPFPTGQHPMA-UHFFFAOYSA-N propylene Natural products CC=C QQONPFPTGQHPMA-UHFFFAOYSA-N 0.000 description 1
- 125000004805 propylene group Chemical group [H]C([H])([H])C([H])([*:1])C([H])([H])[*:2] 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
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- 235000019154 vitamin C Nutrition 0.000 description 1
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- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
A process for preparing 1,3-propanediol by adding fumaric acid to activate microns includes preparing culture media, seed culture and fermenting. It features that the fumaric acid is added to the fermentive culture medium for speeding up the utilization of glycerine by bacterial spawn.
Description
Technical field
The invention belongs to technical field of biochemical industry, particularly provide a kind of external source to add FUMARIC ACID TECH GRADE and promoted microorganism to synthesize 1, the method for ammediol.
Background technology
1, ammediol (being called for short PDO) is a kind of important chemical material, can be used as organic solvent and is applied to industries such as printing ink, printing and dyeing, coating, lubricant, antifreezing agent.1, the topmost purposes of ammediol is as polyester and urethane synthetic monomer, the polytrimethylene terephthalate (PTT) that generates with the terephthalic acid polymerization has particularly shown that than with 1,2-propylene glycol, butyleneglycol, ethylene glycol are the better performance of monomer synthetic polymkeric substance.The tens million of tons of polyethylene terephthalates (PET) of the annual consumption in the whole world at present, and the chemical stability of PTT, biodegradability etc. are suitable with PET, but stain resistance, toughness and rebound resilience and uvioresistant performance etc. are more superior.Ptt fiber also has wear-resisting, advantages such as water-absorbent is low, low static in addition, can be in carpet applications and nylon competition.It also can be used for having the aspect such as non-woven fabrics, engineering plastics, clothes, home decoration, gasket material, fabric of premium properties.PTT is cited as one of 98 years six big petrochemical industry product innovations of the U.S., is considered to the upgrading products that will be PET.
The high-performance of PTT and market potential just were familiar with by people before 50 years, only because of raw material 1, ammediol production technology difficulty is big, cost is high and cause PTT to be difficult to large-scale industrial production, up to now, have only Dupont and Shell two tame transnational companys to adopt traditional chemical synthesis route, with oxyethane or propylene be raw material production only for their synthetic PTT personal 1, ammediol.The shortcoming of chemical synthesis is that by product is many, poor selectivity, and operational condition needs High Temperature High Pressure, and facility investment is huge, and raw material is Nonrenewable resources, and the intermediate product propenal of oxyethane and another route is respectively inflammable and explosive or hypertoxic hazardous substance.Since fermentative Production 1, ammediol selectivity height, and therefore the operational condition gentleness is subjected to special attention in recent years.
Biological synthesis process produces 1, and ammediol is to utilize microorganism disproportionation glycerine to produce.Occurring in nature can be 1 with transformation of glycerol, if the microbial host anaerobism or the facultative anaerobe of ammediol, wherein Cray Bai Shi pneumobacillus (Klebsiella pneumoniae), butyric acid clostridium (Clostridium butyricum) and C. freundii (Citrobacter freundii) have higher 1, the ammediol transformation efficiency, and to glycerine and product 1, ammediol has higher tolerance, therefore has higher development and is worth and application prospect.Microbial metabolism glycerine produces 1, ammediol is mainly regulated and control by Protosol (dha) operon, its metabolism route is divided into oxidation branch road and two branches of reduction branch road: in oxidative pathway, glycerine dehydrogenation under the katalysis of the glycerol dehydrogenase (GDH) that relies on NAD+ changes Protosol (DHA) into, Protosol phosphoric acid under the kinase whose effect of Protosol turns to phosphodihydroxyacetone, and then enters the glycolysis approach and produce metabolism by products such as ATP, NADH and ethanol, acetate, lactic acid; In the reduction approach, glycerine changes the 3-hydroxy propanal under the effect of glycerol dehydratase (GDHt), and the NADH that utilizes oxidative pathway to produce is then depending on 1 of NADH, and the effect of ammediol oxydo-reductase generates 1, ammediol down; By utilizing NADH reduction 3-hydroxy propanal, thalline has been realized the regeneration of NAD+ on physiology.Glycerol dehydrogenase, glycerol dehydratase and 1, ammediol oxydo-reductase are catalysis 1, the key enzyme that ammediol transforms.
At present biological synthesis process produces 1, and ammediol exists that production concentration is low, glycerol conversion yield is low and problem such as production cycle long production intensity is low, thereby lacks the market competitiveness.
In order to promote 1, the production of ammediol mainly contains following several method at present:
1, adopt little oxygen condition bottom fermentation to produce 1, (Wang Jianfeng etc., the klebsiella micro-aerobe fermentation produces 1 to ammediol, the research of ammediol, modern chemical industry, 2001,21 (5): 28-31.Repair will dragon etc., a kind of microbial micro-aerobe fermentation produces 1, the method for ammediol, Chinese patent publication number: CN1348007)
2, adopt and add the auxilliary substrate of glucose work fermentative production 1 under anaerobic, (Zhang Jian etc. are cosubstrate fermentative production 1 with glucose to ammediol, the research of ammediol, modern chemical industry, 2002,22 (6): 32-35.)
3, the method that the employing external source is added reductive agents such as vitamins C or vitamin-E promotes 1, and (Li Chun etc., external source is added reductive agent and promoted thalline to synthesize 1, the method for ammediol, patent publication No.: 1446919A) in ammediol production
Find that by metabolism and enzymatic analysis influence 1, ammediol synthesizes two aspect key factors.The one, with 1, the size of the synthetic key enzyme activity of being correlated with of ammediol comprises glycerol dehydrogenase, glycerol dehydratase and 1, the activity change of ammediol oxydo-reductase in the thalline.Another influence factor is that the ratio of interior NADH content of thalline or NAD/NADH changes.Experimental result shows, 1, and add FUMARIC ACID TECH GRADE in the ammediol fermenting process and can improve 1 effectively, the activity of the synthetic key enzyme of ammediol, reduced the ratio of NAD+/NADH in the thalline simultaneously, thereby made 1, ammediol concentration and production intensity significantly improve.Also not having external source to add FUMARIC ACID TECH GRADE in the present research promotes microorganism to synthesize 1, the report of ammediol.
Summary of the invention
The object of the invention is to provide a kind of external source interpolation FUMARIC ACID TECH GRADE to promote microorganism to synthesize 1, the method for ammediol.Improve in the thalline 1, the activity of the synthetic key enzyme of ammediol reduces the ratio of NAD/NADH simultaneously, thereby significantly improves 1, the concentration of ammediol and production intensity.
The present invention includes the preparation fermention medium, and in fermention medium, insert seed liquor, carry out anaerobism or aerobic fermentation and synthesize 1, the step of ammediol.It is characterized in that: in fermention medium, add FUMARIC ACID TECH GRADE, concentration be 5～25 mmoles/liter.Add the logarithmic phase that choose opportunities is fermented at bacterial classification.
The FUMARIC ACID TECH GRADE of interpolation of the present invention is a kind of external source electron acceptor(EA).
The present invention is applicable to 1, the anaerobism of ammediol and aerobic fermentation process.
The invention has the advantages that and improve in the thalline 1, the activity of the synthetic key enzyme of ammediol reduces the ratio of NAD/NADH simultaneously, thereby significantly improves 1, the concentration of ammediol and production intensity.And, simple to operate, do not increase extra equipment and artificial, only, significantly improved 1 by lower additional input, the production intensity of ammediol obtains higherly 1, and ammediol concentration has reduced fermentation costs.
Embodiment
Example 1:
(1) bacterial classification: Cray Bai Shi bacillus (Klebsiella pneumoniae) M5a1
(2) substratum
The LB slant medium: every liter of volume substratum contains yeast powder 5.0g, peptone 10.0g, agar powder 15.0g, NaCl 10.0g, pH regulator to 7.0.
Seed and fermentation culture based component see Table 1
Table 1
Nutrient media components seed culture medium fermention medium trace element solution
(/l) (/l) (mg/l)
Glycerine 20g 20g ZnCl
270
K
2HPO
4·3H
2O 4.45g 2.225g MnCl
2·4H
2O 100
(NH
4)
2SO
4 2.0g 2.0g H
3BO
3 60
KH
2PO
4 1.3g 0.65g CoCl
2·6H
2O 200
MgSO
4·7H
2O 0.2g 0.2g NiCl
2·6H
2O 25
Yeast powder 1.0g 1.5g NiCl
2H
2O 27.64
The molten 2ml 2ml Na of trace element
2MoO
42H
2O 35
Liquid
CaCO
3 2.0g CuCl
2·H
2O 20
Defoamer 0.1ml CuSO
45H
2O 29.28
Concentrated hydrochloric acid (37%) 0.9ml
(3) training method:
A. seed activation
Picking glycerine is guaranteed bacterial classification one ring transition of Tibetan to the LB slant activation, cultivates down for 30 ℃ and can obtain activated seed in 12 hours.
B. seed culture
Seed culture adopts aerobic to cultivate, and uses the 500ml triangular flask, liquid amount 100ml.30 ℃ of culture temperature, shaking speed 140rpm, incubation time 14-16h.
C. fermentation culture
Fermentation culture is used the 5L fermentor tank, uses in the fermenting process to contain initial glycerol concentration and be the 20g/l fermention medium, and liquid amount 4L, 37 ℃ of culture temperature, feeding nitrogen, air flow 0.2vvm carries out anaerobically fermenting, and the pH value is adjusted to 6.85 with 50%KOH.Fermentor tank mixing speed 150rpm.Wherein control canisters does not add FUMARIC ACID TECH GRADE, and it is 5mM that experimental tank adding FUMARIC ACID TECH GRADE makes it concentration.
(5) fermentation result
Fermentation was carried out 8 hours, and wherein residual glycerol concentration 5.48g/L in the control canisters contains 1 in the fermented liquid, ammediol 7.54g/L, Succinic Acid 0.41g/L, lactic acid 0.88g/L, acetic acid 2.21g/L, 2,3-butyleneglycol 0.03g/L.1, ammediol mole yield 0.55, the glycerine spending rate is 2.06g/ (Lh), 1, ammediol production intensity 0.94g/ (Lh).Add residual glycerol concentration 1.01g/L in the 5mM FUMARIC ACID TECH GRADE jar, contain 1 in the fermented liquid, ammediol 11.10g/L, Succinic Acid 1.12g/L, lactic acid 0.97g/L concentration, acetic acid 3.46g/L, 2,3-butyleneglycol 0.07g/L.1, ammediol mole yield 0.56, the glycerine wear rate is 2.76g/ (Lh), 1, ammediol production intensity 1.28g/ (Lh).As seen, the glycerine spending rate is compared according to improving 34% when adding the 5mM FUMARIC ACID TECH GRADE, and production intensity improves 36%, 1, and ammediol concentration contrasts and improves 47.2%, and 1, the ammediol yield is basic suitable with contrast.
Example 2:
(1) bacterial classification: with example 1
(2) substratum: with example 1
(3) training method:
A. seed activation
Picking glycerine is guaranteed bacterial classification one ring transition of Tibetan to the LB slant activation, cultivates down for 30 ℃ and can obtain activated seed in 12 hours.
B. seed culture
Seed culture adopts aerobic to cultivate, and uses the 500ml triangular flask, liquid amount 100ml.30 ℃ of culture temperature, shaking speed 140rpm, incubation time 14-16h.
C. fermentation culture
Fermentation culture is used the 500mL triangular flask, in the gas bath shaking table, carries out, and 37 ℃ of temperature, shaking speed 100rpm, liquid amount are the 200mL/500mL bottle, inoculum size is 10%, adds 1.g CaCO in the substratum
3Regulate pH.Shake bottle with six layers of gauze wrapping carrying out micro-aerobe fermentation.In order to investigate fumaric influence, 0mM, 5mM, four groups of FUMARIC ACID TECH GRADE concentration tests of 15mM, 25mM are set respectively.
(5) fermentation result
4h is carried out in fermentation altogether, the results are shown in Table 2.
Glycerine consumption and product are synthetic after the table 2 interpolation FUMARIC ACID TECH GRADE
FUMARIC ACID TECH GRADE glycerine consumes synthetic (mol) 1 of product, ammediol yield
(mmole/liter) (mol) (moles/mole)
Glycerine Succinic Acid lactic acid acetate 1, ammediol 2,3-butyleneglycol ethanol
0 64.8 0.6 4.1 do not detect 23.5 5.6 9.1 339.9
5 106.6 4 7.8 do not detect 35.9 6.4 21.1 338.9
15 135.5 9.5 10.1 do not detect 42 7.8 31 317.8
25 143.6 16.7 9.8 0.9 43.1 8.2 34 300.8
As seen, along with the raising of FUMARIC ACID TECH GRADE addition, glycerine spending rate and 1, the production intensity of ammediol all is greatly improved.Interpolation 5,15,25 mmoles/spending rate of glycerine is compared respectively according to improving 65%, 109% and 122% when rising FUMARIC ACID TECH GRADE; 1, the production intensity of ammediol improves 53%, 79% and 83% respectively; 1,3 propylene glycol concentration improves 53%, 79 and 84% respectively.
Claims (3)
1. an external source interpolation FUMARIC ACID TECH GRADE promotes microorganism to synthesize 1, and the method for ammediol comprises: medium preparation, seed culture, and fermentation product 1, the step of ammediol; It is characterized in that: in fermention medium, add FUMARIC ACID TECH GRADE, concentration be 5～25 mmoles/liter; Add the logarithmic phase that choose opportunities is fermented at bacterial classification.
2. the method for claim 1, it is characterized in that: the FUMARIC ACID TECH GRADE of interpolation is a kind of external source electron acceptor(EA).
3. method as claimed in claim 1 or 2 is characterized in that: be applicable to 1, the anaerobism of ammediol and aerobic fermentation process.
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| WO2006128381A1 (en) * | 2005-06-03 | 2006-12-07 | Tsinghua University | Method for preparing 1,3-propanediol by using glycerine as the by-product of the biological diesel oil |
| CN102250968A (en) * | 2011-01-31 | 2011-11-23 | 清华大学 | Method for producing 1, 3-propanediol through fermentation |
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| US5686276A (en) * | 1995-05-12 | 1997-11-11 | E. I. Du Pont De Nemours And Company | Bioconversion of a fermentable carbon source to 1,3-propanediol by a single microorganism |
| CN1204260C (en) * | 2001-04-27 | 2005-06-01 | 大连理工大学 | Microbial micro-aerobe fermentation process of producing 1,3-propylene glycol |
| CN1182249C (en) * | 2003-04-18 | 2004-12-29 | 清华大学 | Method of adding reducing agent from extraneous sources for accelerating thallus to synthesize 1,3-propylene glycol |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2006128381A1 (en) * | 2005-06-03 | 2006-12-07 | Tsinghua University | Method for preparing 1,3-propanediol by using glycerine as the by-product of the biological diesel oil |
| CN102250968A (en) * | 2011-01-31 | 2011-11-23 | 清华大学 | Method for producing 1, 3-propanediol through fermentation |
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