CN114807249B - Catalytic CO 2 Multi-enzyme cascade pathway for synthesizing 2C or 4C compounds - Google Patents
Catalytic CO 2 Multi-enzyme cascade pathway for synthesizing 2C or 4C compounds Download PDFInfo
- Publication number
- CN114807249B CN114807249B CN202210306243.2A CN202210306243A CN114807249B CN 114807249 B CN114807249 B CN 114807249B CN 202210306243 A CN202210306243 A CN 202210306243A CN 114807249 B CN114807249 B CN 114807249B
- Authority
- CN
- China
- Prior art keywords
- glycolaldehyde
- methanol
- synthesizing
- erythrose
- synthase
- 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.)
- Active
Links
- 230000002194 synthesizing effect Effects 0.000 title claims abstract description 76
- 150000001875 compounds Chemical class 0.000 title claims abstract description 45
- 230000003197 catalytic effect Effects 0.000 title claims description 21
- 230000037361 pathway Effects 0.000 title abstract description 29
- WGCNASOHLSPBMP-UHFFFAOYSA-N Glycolaldehyde Chemical compound OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims abstract description 272
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 253
- 102000004190 Enzymes Human genes 0.000 claims abstract description 83
- 108090000790 Enzymes Proteins 0.000 claims abstract description 83
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 claims abstract description 80
- 108010025188 Alcohol oxidase Proteins 0.000 claims abstract description 79
- YTBSYETUWUMLBZ-UHFFFAOYSA-N D-Erythrose Natural products OCC(O)C(O)C=O YTBSYETUWUMLBZ-UHFFFAOYSA-N 0.000 claims abstract description 72
- YTBSYETUWUMLBZ-IUYQGCFVSA-N D-erythrose Chemical compound OC[C@@H](O)[C@@H](O)C=O YTBSYETUWUMLBZ-IUYQGCFVSA-N 0.000 claims abstract description 61
- 238000000034 method Methods 0.000 claims abstract description 46
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 39
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 37
- 102000003960 Ligases Human genes 0.000 claims abstract description 34
- 108090000364 Ligases Proteins 0.000 claims abstract description 34
- 101710088194 Dehydrogenase Proteins 0.000 claims abstract description 27
- 238000006555 catalytic reaction Methods 0.000 claims abstract description 18
- 230000009466 transformation Effects 0.000 claims abstract description 13
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims description 144
- 238000006243 chemical reaction Methods 0.000 claims description 42
- 239000000758 substrate Substances 0.000 claims description 21
- 101000928995 Caenorhabditis elegans Putative deoxyribose-phosphate aldolase Proteins 0.000 claims description 18
- 102100037802 Deoxyribose-phosphate aldolase Human genes 0.000 claims description 18
- 108090000623 proteins and genes Proteins 0.000 claims description 16
- 108010068673 Glycolaldehyde dehydrogenase Proteins 0.000 claims description 14
- 108010081577 aldehyde dehydrogenase (NAD(P)+) Proteins 0.000 claims description 12
- 102000016938 Catalase Human genes 0.000 claims description 10
- 108010053835 Catalase Proteins 0.000 claims description 10
- AYEKOFBPNLCAJY-UHFFFAOYSA-O thiamine pyrophosphate Chemical compound CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N AYEKOFBPNLCAJY-UHFFFAOYSA-O 0.000 claims description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L magnesium sulphate Substances [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 7
- 239000011678 thiamine pyrophosphate Substances 0.000 claims description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims 4
- 108020002663 Aldehyde Dehydrogenase Proteins 0.000 claims 1
- 102000005369 Aldehyde Dehydrogenase Human genes 0.000 claims 1
- 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 claims 1
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 abstract description 174
- 229910052799 carbon Inorganic materials 0.000 abstract description 19
- 239000002994 raw material Substances 0.000 abstract description 16
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 15
- 238000004519 manufacturing process Methods 0.000 abstract description 8
- 238000013459 approach Methods 0.000 abstract description 7
- 108030005073 Glycolate dehydrogenases Proteins 0.000 abstract description 6
- 238000006386 neutralization reaction Methods 0.000 abstract description 6
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 12
- 230000005611 electricity Effects 0.000 description 12
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 10
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 10
- -1 preferably Chemical compound 0.000 description 8
- 239000001569 carbon dioxide Substances 0.000 description 6
- 229910002092 carbon dioxide Inorganic materials 0.000 description 6
- 239000003054 catalyst Substances 0.000 description 6
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion 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-N 0.000 description 5
- 229950006238 nadide Drugs 0.000 description 5
- 230000035484 reaction time Effects 0.000 description 5
- 235000008170 thiamine pyrophosphate Nutrition 0.000 description 5
- 239000007853 buffer solution Substances 0.000 description 4
- LEQAOMBKQFMDFZ-UHFFFAOYSA-N glyoxal Chemical compound O=CC=O LEQAOMBKQFMDFZ-UHFFFAOYSA-N 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 101100001035 Gluconobacter oxydans (strain 621H) adhA gene Proteins 0.000 description 3
- 101100162138 Gluconobacter oxydans (strain 621H) adhB gene Proteins 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000008055 phosphate buffer solution Substances 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- KMTRUDSVKNLOMY-UHFFFAOYSA-N Ethylene carbonate Chemical compound O=C1OCCO1 KMTRUDSVKNLOMY-UHFFFAOYSA-N 0.000 description 2
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- AYJRCSIUFZENHW-UHFFFAOYSA-L barium carbonate Chemical compound [Ba+2].[O-]C([O-])=O AYJRCSIUFZENHW-UHFFFAOYSA-L 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000007036 catalytic synthesis reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 229940015043 glyoxal Drugs 0.000 description 2
- 239000005431 greenhouse gas Substances 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 2
- 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 2
- 229910000510 noble metal Inorganic materials 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 150000005846 sugar alcohols Chemical class 0.000 description 2
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 206010056474 Erythrosis Diseases 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 1
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical compound [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 description 1
- 206010000059 abdominal discomfort Diseases 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 125000003172 aldehyde group Chemical group 0.000 description 1
- 229920000180 alkyd Polymers 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000013064 chemical raw material Substances 0.000 description 1
- FOCAUTSVDIKZOP-UHFFFAOYSA-N chloroacetic acid Chemical compound OC(=O)CCl FOCAUTSVDIKZOP-UHFFFAOYSA-N 0.000 description 1
- 229940106681 chloroacetic acid Drugs 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000007323 disproportionation reaction Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000009713 electroplating Methods 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 229940050410 gluconate Drugs 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000005984 hydrogenation reaction Methods 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 239000003444 phase transfer catalyst Substances 0.000 description 1
- 230000001699 photocatalysis Effects 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- FIJPWGLOBMXXSF-UHFFFAOYSA-M potassium;2-hydroxyacetate Chemical compound [K+].OCC([O-])=O FIJPWGLOBMXXSF-UHFFFAOYSA-M 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229910052707 ruthenium Inorganic materials 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229960002363 thiamine pyrophosphate Drugs 0.000 description 1
- YXVCLPJQTZXJLH-UHFFFAOYSA-N thiamine(1+) diphosphate chloride Chemical compound [Cl-].CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N YXVCLPJQTZXJLH-UHFFFAOYSA-N 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/42—Hydroxy-carboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention relates to a method for catalyzing CO 2 Multiple enzyme cascade pathway for synthesizing 2C or 4C compounds in CO 2 Glycolic acid and ethylene glycol and D-erythrose are synthesized by a multienzyme cascade as raw materials. The route specifically comprises the following steps: (1) Methanol synthesis of glycolic acid is catalyzed by a cascade of methanol oxidase, glycolaldehyde synthase and glycolate dehydrogenase; (2) Synthesizing ethylene glycol by catalyzing methanol through cascade connection of methanol oxidase, glycolaldehyde synthetase and ethylene glycol dehydrogenase; (3) Catalyzing methanol synthesis of D-erythrose by a multi-enzyme cascade of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase; wherein methanol catalyzes CO by photo, electro or multienzyme 2 And (3) transformation. The above approach is based on the background of carbon neutralization and carbon arrival peaks, realizing CO 2 The multi-enzyme cascade catalysis for synthesizing the 2C compound and the 4C compound provides a new idea for synthesizing the multi-carbon compound from the 1C compound, and opens up a new way for the green biological production of the multi-carbon high-added-value compound.
Description
Technical Field
The invention belongs to the technical field of biocatalysis application, and relates to a method for catalyzing CO 2 A multi-enzyme cascade pathway for the synthesis of 2C or 4C compounds.
Background
Glycolic acid is also called glycolic acid and glycollic acid, is an important organic synthesis intermediate and chemical product, and is widely applied to the industries of organic synthesis, cleaning, electroplating, spinning, leather, sterilization and the like. In addition, the glycolic acid polymer has biodegradability, solves the problem of difficult degradation of the traditional plastic products, and can be widely applied to medicine, packaging and other fields.
Ethylene glycol, also called glycol and 1, 2-ethylene glycol, is the simplest dihydric alcohol, is an important organic chemical raw material, and is widely applied to the fields of producing polyesters, alkyd resins, lubricants, paint, adhesives, surfactants and the like. The ethylene glycol can be biodegraded, and the polymer also has biodegradability, and the ethylene glycol is taken as a monomer to synthesize the polymer, so that the problem that the traditional plastic product is difficult to degrade is solved.
D-erythritol, a four-carbon sugar with a reducing aldehyde group, is unstable in structure under conventional conditions, and erythritol, which is a natural healthy sugar alcohol and is white crystals, can be further prepared through hydrogenation reaction and is widely existing in fruits and fermented foods in a low content. The D-erythritol caloric value is the lowest among all sugar alcohols and does not cause gastrointestinal discomfort under normal eating conditions and does not cause adverse gastrointestinal reactions. D-erythrose is currently considered as one of important platform compounds, a series of products with wide application range can be obtained through chiral conversion and achiral conversion, and the D-erythrose can also be used as an intermediate of fine chemical industry and pharmacy, and has great economic value and wide application prospect.
The traditional glycollic acid preparation method mainly comprises (1) taking formaldehyde and CO as raw materials, synthesizing glycollic acid through carbonyl of formaldehyde under the catalysis of strong acid; (2) Chloroacetic acid is taken as a raw material, and glycolic acid is prepared by hydrolysis under the action of barium carbonate or calcium carbonate serving as a catalyst; (3) Glyoxal is used as raw material, and is oxidized to synthesize glycollic acid under alkaline condition. Chinese patent CN102584566a discloses a method for preparing glycolic acid from glyoxal as substrate, potassium glycolate through disproportionation reaction under the action of potassium hydroxide and phase transfer catalyst, and glycolic acid through acidification. Chinese patent CN105085227a provides a method of hydrolyzing methyl glycolate using a hydrolysis reactor, then separating by a methanol refining tower and concentrating by a concentration tower to obtain glycolic acid. The method is to prepare the glycollic acid by using a chemical method, and has the advantages of high toxicity, harsh conditions, long synthesis period and low product purity.
The current ethylene glycol preparation method mainly comprises the following steps: (1) catalytic hydration method: ethylene oxide and water are used as substrates, and the catalyst is synthesized under certain temperature and pressure conditions. The method has the advantages of long process flow, complex equipment and high energy consumption. (2) ethylene carbonate method: first CO is reacted with a catalyst 2 And ethylene oxide reacts to produce ethylene carbonate, and then ethylene glycol is obtained through hydrolysis, so that the selectivity is high, the energy consumption is high, and the conditions are severe. (3) synthesis gas process: the glycol is synthesized by high temperature and high pressure and the action of noble metal catalyst, and the conditions are harsh. Patent (CN 102649686 a) discloses a method for synthesizing glycollate and then hydrolyzing to obtain glycol by using oxalic ester and hydrogen as raw materials and noble metal as a catalyst, which improves the selectivity of glycol.
At present, D-erythrose has the following methods: patent (CN 201410854859.9) reports a method for preparing D-erythrose by using biomass sugar, wherein the invention takes biomass sugar as a raw material and catalyzes the biomass sugar to obtain the D-erythrose under an alkaline environment under an aqueous phase condition. Patent (CN 97121416.6) discloses a process for the preparation of D-erythrose by contacting an aqueous solution of gluconate with hydrogen peroxide in the presence of a salt of a metal selected from cobalt, nickel and ruthenium.
At present, few methods for synthesizing glycolic acid, ethylene glycol and D-erythrose by an enzymatic method are reported. In the form of CO 2 The pathway of synthesizing glycolic acid, ethylene glycol and D-erythrose by multienzyme cascade catalysis is also hardly reported as a raw material. Development of a green and efficient CO 2 The approach to the synthesis of 2C and 4C compounds is a long felt but unsolved problem.
Disclosure of Invention
The invention aims to provide a catalyst CO based on the background of carbon neutralization carbon peak 2 A multi-enzyme cascade pathway for the synthesis of 2C or 4C compounds. The path is carried out by CO 2 As raw materials, two 2C compounds-glycolic acid, ethylene glycol and a 4C compound-D-erythrose are obtained through three multi-enzyme cascade catalytic pathways. Wherein CO is 2 Catalyzed by light, electricity or enzymesIs converted into methanol. The above approach provides a new idea for synthesizing multi-carbon compounds from 1C compounds.
To this end, the invention provides a catalytic CO 2 A multi-enzyme cascade pathway for the synthesis of 2C or 4C compounds comprising: multienzyme cascade catalysis of CO 2 Pathway for synthesizing glycollic acid and multienzyme cascade catalysis CO 2 Pathway for synthesizing glycol or multienzyme cascade catalysis CO 2 A pathway for synthesizing D-erythrose.
According to an embodiment of the first aspect of the invention, the multienzyme cascade catalyzes CO 2 The pathway for synthesizing glycolic acid comprises using 3 enzymes of methanol oxidase, glycolaldehyde synthase and glycolaldehyde dehydrogenase to catalyze the synthesis of glycolic acid from a 2C compound, wherein methanol catalyzes CO by light, electricity or multiple enzymes 2 And (3) transformation.
In particular, the multienzyme cascade catalyzes CO 2 Pathways for synthesizing glycolic acid include:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) Glycolaldehyde is catalyzed by glycolaldehyde dehydrogenase to synthesize glycolaldehyde;
in the invention, the methanol oxidase is a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; the glycolaldehyde synthase is a related enzyme with the function of catalyzing formaldehyde to synthesize glycolaldehyde; the glycollic acid dehydrogenase is a related enzyme with the function of catalyzing glycolaldehyde to synthesize glycollic acid.
Preferably, the methanol oxidase comprises an AOX enzyme; the glycolaldehyde synthase includes a GALS enzyme; the glycolate dehydrogenase includes an acetaldehyde dehydrogenase ALDH-BL21 and/or an acetaldehyde dehydrogenase ALDH-11300.
According to an embodiment of the second aspect of the invention, the multienzyme cascade catalyzes CO 2 The synthesis of glycol comprises using 3 enzymes of methanol oxidase, glycolaldehyde synthase and glycol dehydrogenase to catalyze the synthesis of glycol from methanol to 2C compound, wherein methanol catalyzes CO by light, electricity or multiple enzymes 2 And (3) transformation.
In particular, the multienzyme cascade catalyzes CO 2 The synthetic routes to ethylene glycol include:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) Synthesizing glycol by catalyzing glycolaldehyde with glycol dehydrogenase;
in the invention, the methanol oxidase is a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; the glycolaldehyde synthase is a related enzyme with the function of catalyzing formaldehyde to synthesize glycolaldehyde; the glycol dehydrogenase is a related enzyme with the function of catalyzing glycolaldehyde to synthesize glycol;
preferably, the methanol oxidase comprises an AOX enzyme; the glycolaldehyde synthase includes a GALS enzyme; the ethylene glycol dehydrogenase comprises one or more of alcohol dehydrogenase GOX1067, alcohol dehydrogenase GOX1068, acetaldehyde dehydrogenase GOX2018 and alcohol dehydrogenase GOX 0313.
According to an embodiment of the third aspect of the invention, the multienzyme cascade catalyzes CO 2 The pathway for synthesizing D-erythrose includes 3 enzyme cascade catalysis of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase to synthesize D-erythrose as 4C compound, wherein methanol catalyzes CO via photo, electro or multienzyme 2 And (3) transformation.
In particular, the multienzyme cascade catalyzes CO 2 The synthetic pathways for D-erythrose include:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) D-erythrose is synthesized by catalyzing glycolaldehyde by using D-erythrose synthetase.
In the invention, the methanol oxidase is a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; the glycolaldehyde synthase is a related enzyme with the function of catalyzing formaldehyde to synthesize glycolaldehyde; the D-erythrose synthetase is a related enzyme with the function of catalyzing glycolaldehyde to synthesize D-erythrose;
preferably, the methanol oxidase comprises an AOX enzyme; the glycolaldehyde synthase includes a GALS enzyme; the D-erythrose synthetase comprises one or more of 2-deoxy-D-ribose 5-phosphate aldolase DERAEco, 2-deoxy-D-ribose 5-phosphate aldolase DERAYer, 2-deoxy-D-ribose 5-phosphate aldolase DERASa, 2-deoxy-D-ribose 5-phosphate aldolase DERAMet and 2-deoxy-D-ribose 5-phosphate aldolase DERAPyc.
The invention also provides a multi-enzyme cascade catalytic CO 2 A method for synthesizing a 2C or 4C compound comprising a multienzyme cascade catalyzing CO 2 Synthesis of glycolic acid, multienzyme cascade catalysis CO 2 Synthesis of ethylene glycol or multienzyme cascade catalysis of CO 2 Synthesizing D-erythrose.
According to an embodiment of the fourth aspect of the invention, the multienzyme cascade catalyzes CO 2 The method for synthesizing the glycollic acid comprises the steps of realizing the synthesis of glycollic acid of a methanol-to-2C compound by using 3 enzyme cascade catalysis of methanol oxidase, glycolaldehyde synthase and glycollic acid dehydrogenase, wherein the methanol catalyzes CO by light, electricity or multiple enzymes 2 Obtaining by transformation;
in particular, the multienzyme cascade catalyzes CO 2 The method for synthesizing the glycollic acid comprises the following steps:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) Glycolaldehyde is catalyzed by glycolaldehyde dehydrogenase to synthesize glycolaldehyde;
preferably, the methanol oxidase is a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; the glycolaldehyde synthase is a related enzyme with the function of catalyzing formaldehyde to synthesize glycolaldehyde; the glycollic acid dehydrogenase is a related enzyme with the function of catalyzing glycolaldehyde to synthesize glycollic acid;
further preferably, the methanol oxidase comprises an AOX enzyme; the glycolaldehyde synthase includes a GALS enzyme; the glycolate dehydrogenase includes an acetaldehyde dehydrogenase ALDH-BL21 and/or an acetaldehyde dehydrogenase ALDH-11300.
In some embodiments of the inventionThe multienzyme cascade catalyzes CO 2 The method for synthesizing the glycollic acid takes methanol as a raw material, and utilizes 3 enzymes of methanol oxidase, glycolaldehyde synthase and glycolaldehyde dehydrogenase to catalyze the reaction for synthesizing the glycollic acid at the temperature of 4-80 ℃; the pH of the reaction is 3-14; the mass of the methanol oxidase, glycolaldehyde synthase and glycolaldehyde dehydrogenase is (0.1-10): (0.1-10): (0.1-10).
According to an embodiment of the fifth aspect of the invention, the multienzyme cascade catalyzes CO 2 The method for synthesizing glycol comprises using 3 enzymes of methanol oxidase, glycolaldehyde synthase and glycol dehydrogenase to realize synthesis of glycol from methanol to 2C compound, wherein the methanol catalyzes CO by light, electricity or multiple enzymes 2 Obtaining by transformation;
in particular, the multienzyme cascade catalyzes CO 2 The method for synthesizing the ethylene glycol comprises the following steps:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) Synthesizing glycol by catalyzing glycolaldehyde with glycol dehydrogenase;
preferably, the methanol oxidase is a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; the glycolaldehyde synthase is a related enzyme with the function of catalyzing formaldehyde to synthesize glycolaldehyde; the glycol dehydrogenase is a related enzyme with the function of catalyzing glycolaldehyde to synthesize glycol;
further preferably, the methanol oxidase comprises an AOX enzyme; the glycolaldehyde synthase includes a GALS enzyme; the ethylene glycol dehydrogenase comprises one or more of alcohol dehydrogenase GOX1067, alcohol dehydrogenase GOX1068, acetaldehyde dehydrogenase GOX2018 and alcohol dehydrogenase GOX 0313.
In some embodiments of the invention, the multi-enzyme cascade catalyzes CO 2 The method for synthesizing glycol takes methanol as raw material, and utilizes 3 enzymes of methanol oxidase, glycolaldehyde synthase and glycol dehydrogenase to catalyze and synthesize glycol at 4-80 ℃; the pH of the reaction is 3-14; quality of methanol oxidase, glycolaldehyde synthase and glycol dehydrogenaseThe ratio is (0.1-10): (0.1-10): (0.1-10).
According to an embodiment of the sixth aspect of the invention, the multienzyme cascade catalyzes CO 2 The method for synthesizing D-erythrose comprises using 3 enzymes of methanol oxidase, glycolaldehyde synthetase and D-erythrose synthetase to realize synthesis of D-erythrose from methanol to 4C compound, wherein the methanol catalyzes CO by light, electricity or multiple enzymes 2 Obtaining by transformation;
in particular, the multienzyme cascade catalyzes CO 2 The method for synthesizing the D-erythrose comprises the following steps:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) Catalyzing glycolaldehyde to synthesize D-erythrose by using D-erythrose synthetase;
preferably, the methanol oxidase is a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; the glycolaldehyde synthase is a related enzyme with the function of catalyzing formaldehyde to synthesize glycolaldehyde; the D-erythrose synthetase is a related enzyme with the function of catalyzing glycolaldehyde to synthesize D-erythrose;
further preferably, the methanol oxidase comprises an AOX enzyme; the glycolaldehyde synthase includes a GALS enzyme; the D-erythrose synthetase comprises one or more of 2-deoxy-D-ribose 5-phosphate aldolase DERAEco, 2-deoxy-D-ribose 5-phosphate aldolase DERAYer, 2-deoxy-D-ribose 5-phosphate aldolase DERASa, 2-deoxy-D-ribose 5-phosphate aldolase DERAMet and 2-deoxy-D-ribose 5-phosphate aldolase DERAPyc.
In some embodiments of the invention, the multi-enzyme cascade catalyzes CO 2 The method for synthesizing the D-erythrose takes methanol as a raw material, and 3 enzymes of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase are used for cascading and catalyzing to synthesize the D-erythrose at the temperature of 4-80 ℃; the pH of the reaction is 3-14; the ratio of 3 enzymes of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase is (0.1-10): (0.1-10): (0.1-10).
The invention provides a method for catalyzing CO 2 Synthesis of 2C or 4C CompoundsIs a multi-enzyme cascade pathway of (2) in CO 2 As raw materials, two 2C compounds, glycolic acid and ethylene glycol and one 4C compound D-erythrose, were synthesized by a multienzyme cascade, comprising in particular 3 pathways: (1) Methanol synthesis of glycolic acid is catalyzed by a cascade of methanol oxidase, glycolaldehyde synthase and glycolate dehydrogenase; (2) Synthesizing ethylene glycol by catalyzing methanol through cascade connection of methanol oxidase, glycolaldehyde synthetase and ethylene glycol dehydrogenase; (3) Catalyzing methanol synthesis of D-erythrose by a multi-enzyme cascade of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase; wherein methanol can catalyze CO by light, electricity or multiple enzymes 2 And (3) transformation. The path of the invention realizes CO based on the background of carbon neutralization and carbon peak 2 The invention provides a new idea for synthesizing multi-carbon compounds from 1C compounds and opens up a new way for green biological production of multi-carbon high-added-value compounds.
Drawings
The invention is described in further detail below with reference to the accompanying drawings:
FIG. 1 shows the catalytic CO in the present invention 2 A multi-enzyme cascade pathway for the synthesis of 2C or 4C compounds.
FIG. 2 shows the progress of a reaction for catalyzing the production of glycolic acid by a cascade of methanol oxidase, glycolaldehyde synthase and glycolate dehydrogenase.
FIG. 3 shows the reaction sequence of a cascade catalyzed ethylene glycol production by methanol oxidase, glycolaldehyde synthase and ethylene glycol dehydrogenase.
FIG. 4 shows the progress of a reaction of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase in cascade catalysis to produce D-erythrose.
Detailed Description
In order that the invention may be readily understood, the invention will be described in detail below with reference to the accompanying drawings. Before the present invention is described in detail, it is to be understood that this invention is not limited to particular embodiments described. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.
Unless defined otherwise, all terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present invention, the preferred methods and materials are now described.
I terminology
The term "TPP" as used herein means thiamine pyrophosphate, "NADH" means reduced coenzyme I, "NAD + "refers to oxidized coenzyme I.
The terms "protein" and "protein" as used herein may be used interchangeably.
The term carbon neutralization (carbon neutrality) refers to the total emission amount of carbon dioxide or greenhouse gases generated by countries, enterprises, products, activities or individuals directly or indirectly in a certain time, and the carbon dioxide or the greenhouse gases generated by the countries, enterprises, products, activities or individuals are offset by modes of tree planting, energy saving, emission reduction, carbon dioxide recovery and the like, so that positive and negative offset is realized, and the relative zero emission is achieved.
The term "peak of carbon" (peak carbon dioxide emissions) as used herein means that at a certain point in time, the carbon dioxide emissions no longer increase to peak and then fall back gradually. The peak reaching carbon is a historical inflection point of the carbon dioxide emission from increasing to decreasing, marks the unhooking of the carbon emission and the economic development, and the peak reaching targets comprise peak reaching years and peaks.
II. Embodiment
As described above, few methods for synthesizing glycolic acid, ethylene glycol and D-erythrose by an enzymatic method have been reported. In the form of CO 2 The pathway of synthesizing glycolic acid, ethylene glycol and D-erythrose by multienzyme cascade catalysis is also hardly reported as a raw material. Development of a green and efficient CO 2 The approach to the synthesis of 2C and 4C compounds is a long felt but unsolved problem. In view of this, the invention is efficient for green CO 2 Methods for synthesizing 2C and 4C compounds have been extensively studied.
Based on the background of carbon neutralization carbon peak, the invention researches and discovers and constructs a catalytic CO 2 Multi-enzyme cascade for synthesizing 2C or 4C compoundsDiameter, the pathway is as CO 2 As a starting material, two 2C compounds- -glycolic acid and ethylene glycol and one 4C compound D-erythrose- -were synthesized by a multienzyme cascade, as shown in FIG. 1.
The first catalytic CO constructed by the invention 2 The multienzyme cascade path for synthesizing 2C or 4C compounds is multienzyme cascade catalytic CO 2 A pathway for synthesizing glycolic acid comprising effecting synthesis of glycolaldehyde, a methanol to 2C compound, using a combination of 3 enzymes, methanol oxidase, glycolaldehyde synthase and glycolaldehyde dehydrogenase; wherein the methanol catalyzes CO by photo, electro or multienzyme 2 And (3) transformation. The route specifically comprises the following steps: (1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde; (2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase; (3) Glycolaldehyde synthesis is catalyzed by glycolaldehyde dehydrogenase (see FIG. 1).
Corresponding to the above approach, the present invention also provides a multienzyme cascade catalytic CO 2 A method of synthesizing glycolic acid comprising the steps of: (1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde; (2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase; (3) Glycolaldehyde is catalyzed by glycolaldehyde dehydrogenase to synthesize glycolaldehyde.
In the invention, the methanol oxidase refers to a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; particularly preferably, the methanol oxidase is an AOX enzyme.
In the invention, the glycolaldehyde synthetase refers to a related enzyme with a function of catalyzing formaldehyde to synthesize glycolaldehyde; particularly preferably, the glycolaldehyde synthase is a GALS enzyme.
In the present invention, the glycollic acid dehydrogenase refers to a related enzyme having a function of catalyzing glycolaldehyde to synthesize glycollic acid; particularly preferably, the glycolate dehydrogenase is an acetaldehyde dehydrogenase ALDH-BL21 and/or an acetaldehyde dehydrogenase ALDH-11300.
In some embodiments of the invention, the multi-enzyme cascade catalyzes CO 2 The method for synthesizing the glycollic acid takes methanol as a raw material, and utilizes 3 enzymes of methanol oxidase, glycolaldehyde synthase and glycolaldehyde dehydrogenase to catalyze the reaction for synthesizing the glycollic acid at the temperature of 4-80 ℃; pH of the reaction3-14; the mass of the methanol oxidase, glycolaldehyde synthase and glycolaldehyde dehydrogenase is (0.1-10): (0.1-10): (0.1-10).
The second catalytic CO constructed by the invention 2 The multienzyme cascade path for synthesizing 2C or 4C compounds is that the multienzyme cascade catalyzes CO 2 A pathway for synthesizing ethylene glycol using 3 enzyme combinations of methanol oxidase, glycolaldehyde synthase and ethylene glycol dehydrogenase to effect synthesis of methanol to 2C compound ethylene glycol, wherein methanol can catalyze CO by light, electricity or multiple enzymes 2 And (3) transformation. The route specifically comprises the following steps: (1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde; (2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase; (3) Glycolaldehyde is catalyzed by glycol dehydrogenase to synthesize glycol (see FIG. 1).
Corresponding to the above approach, the present invention also provides a multienzyme cascade catalytic CO 2 A method of synthesizing ethylene glycol comprising the steps of: (1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde; (2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase; (3) Glycol dehydrogenase is used for catalyzing glycolaldehyde to synthesize glycol.
In the invention, the methanol oxidase refers to a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; particularly preferably, the methanol oxidase is an AOX enzyme.
In the invention, the glycolaldehyde synthetase refers to a related enzyme with a function of catalyzing formaldehyde to synthesize glycolaldehyde; particularly preferably, the glycolaldehyde synthase is a GALS enzyme.
In the invention, the glycol dehydrogenase refers to a related enzyme with the function of catalyzing glycolaldehyde to synthesize glycol; particularly preferably, the glycol dehydrogenase is one or more of alcohol dehydrogenase GOX1067, alcohol dehydrogenase GOX1068, acetaldehyde dehydrogenase GOX2018 and alcohol dehydrogenase GOX 0313.
In some embodiments of the invention, the multi-enzyme cascade catalyzes CO 2 The method for synthesizing glycol takes methanol as raw material, and utilizes 3 enzymes of methanol oxidase, glycolaldehyde synthase and glycol dehydrogenase to catalyze and synthesize glycol at 4-80 ℃; reverse-rotationThe pH is 3-14; the mass ratio of the methanol oxidase, the glycolaldehyde synthase and the glycol dehydrogenase is (0.1-10): (0.1-10): (0.1-10).
Third catalytic CO constructed by the invention 2 The multienzyme cascade path for synthesizing 2C or 4C compounds is that the multienzyme cascade catalyzes CO 2 A pathway for synthesizing D-erythrose, which utilizes 3 enzyme combinations of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase to realize the synthesis of the compound D-erythrose from methanol to 4C, wherein the methanol can catalyze CO by light, electricity or multiple enzymes 2 And (3) transformation. The route specifically comprises the following steps: (1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde; (2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase; (3) The glycolaldehyde synthesis of D-erythrose is catalyzed by D-erythrose synthase (see FIG. 1).
Corresponding to the above approach, the present invention also provides a multienzyme cascade catalytic CO 2 A method of synthesizing D-erythrose comprising the steps of: (1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde; (2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase; (3) D-erythrose is synthesized by catalyzing glycolaldehyde by using D-erythrose synthetase.
In the invention, the methanol oxidase refers to a related enzyme with the function of catalyzing methanol to synthesize formaldehyde; particularly preferably, the methanol oxidase is an AOX enzyme.
In the invention, the glycolaldehyde synthetase refers to a related enzyme with a function of catalyzing formaldehyde to synthesize glycolaldehyde; particularly preferably, the glycolaldehyde synthase is a GALS enzyme.
In the present invention, the D-erythrose synthase means an enzyme having a function of catalyzing glycolaldehyde to synthesize D-erythrose, and particularly preferably, the D-erythrose synthase is one or more of 2-deoxy-D-ribose 5-phosphate aldolase deroeo, 2-deoxy-D-ribose 5-phosphate aldolase DERAYer, 2-deoxy-D-ribose 5-phosphate aldolase DERASa, 2-deoxy-D-ribose 5-phosphate aldolase DERAMet, and 2-deoxy-D-ribose 5-phosphate aldolase derayc.
In some embodiments of the invention, the multi-enzyme cascade catalyzes CO 2 Synthesis of D-erythrosineThe method for synthesizing the sugar takes methanol as a raw material, and utilizes 3 enzymes of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase to catalyze and synthesize the D-erythrose in a cascading way at the temperature of 4-80 ℃; the pH of the reaction is 3-14; the ratio of 3 enzymes of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase is (0.1-10): (0.1-10): (0.1-10).
It will be appreciated by those skilled in the art that the catalytic CO described in the present invention 2 The multienzyme cascade pathway for synthesizing 2C or 4C compounds is referred to as CO 2 For starting materials, CO is catalyzed by light, electricity or multienzymes 2 Converting to obtain methanol, and then taking methanol as a substrate to catalyze CO in multi-enzyme cascade 2 Pathway for synthesizing glycollic acid and multienzyme cascade catalysis CO 2 Pathway for synthesizing glycol or multienzyme cascade catalysis CO 2 Pathways for the synthesis of D-erythrose 2C or 4C compounds are synthesized as shown in figure 1. Thus, the catalytic CO provided in the present invention 2 The multienzyme cascade pathway for the synthesis of 2C or 4C compounds can also be understood as catalytic CO 2 A multi-enzyme cascade pathway that converts to methanol and catalyzes the synthesis of 2C or 4C compounds from methanol; accordingly, the multienzyme cascade catalysis CO provided in the present invention 2 Methods for synthesizing 2C or 4C compounds can also be understood as catalytic CO 2 A method for converting the methanol into methanol and catalyzing the methanol to synthesize 2C or 4C compounds by a multi-enzyme cascade; based on the realization of carbon neutralization and carbon peak reaching, CO is realized 2 The multi-enzyme cascade catalysis for synthesizing the 2C compound and the 4C compound provides a new idea for synthesizing the multi-carbon compound from the 1C compound, and opens up a new way for the green biological production of the multi-carbon high-added-value compound.
In the present invention, for the catalysis of CO by light, electricity or multiple enzymes 2 The method for obtaining methanol by conversion is not particularly limited as long as it is capable of catalyzing CO by light, electricity or multiple enzymes 2 Conversion to methanol is sufficient, for example, reference DOI:10.1002/anie.201812773 and DOI: the method described in 10.1002/anie.201808964 is by photocatalytic or electrocatalytic CO 2 The conversion gives methanol.
The detection method and the detection instrument in the invention are as follows:
(1) The yield of the product (e.g., glycolic acid, ethylene glycol, and D-erythrose) was determined using HPLC U3000 type high performance liquid chromatograph (Thermo Fisher Scientific).
Examples
In order that the invention may be more readily understood, the invention will be further described in detail with reference to the following examples, which are given by way of illustration only and are not limiting in scope of application. The starting materials or components used in the present invention may be prepared by commercial or conventional methods unless specifically indicated. The specific experimental methods not mentioned in the following examples are generally carried out according to conventional experimental methods.
Example 1: catalytic synthesis of glycollic acid by cascade of methanol oxidase, glycolaldehyde synthase and glycollic acid dehydrogenase
The reaction system: the total volume of the reaction system is 3mL, and the buffer solution is phosphate buffer solution PBS. After reacting for 1-12h at 4-80 ℃, the amount of glycolic acid produced is detected by HPLC.
The reaction equation is shown in formula (1):
the total volume of the reaction system was 3mL, the buffer was 0.05M, PBS with pH=8.0, 50mM methanol, 2mM anhydrous magnesium sulfate, 0.1mM TPP,25mM NAD +,3mg methanol oxidase AOX,0.3mg catalase CAT,6mg glycolaldehyde synthase GALS were added to the reaction system, after 6 hours reaction at 35℃3mg acetaldehyde dehydrogenase ALDH-BL21 was added, and after 0.5 hours continuous reaction at 35℃glycolic acid was detected, 0.33g/L glycolic acid was obtained. The graph of the glycolic acid production reaction progress is shown in FIG. 2 (note: abscissa reaction time refers to the reaction time calculated after addition of ALDH-BL 21).
Methanol oxidase AOX and catalase CAT were all purchased from Sigma. The glycolaldehyde synthase GALS gene was obtained from the literature (DOI No. 10.1038/s41467-019-09095 of this literature) based on which it was obtained by Huada gene synthesis. The acetaldehyde dehydrogenase ALDH-BL21 was obtained from a patent (the patent number: 202111282888.9, genBank accession number: CAQ31929.1 of the gene encoding the enzyme) (based on this patent, from Huada gene synthesis).
Example 2: catalytic synthesis of ethylene glycol by cascade of methanol oxidase, glycolaldehyde synthase and ethylene glycol dehydrogenase
The reaction system: the total volume of the reaction system is 3mL, and the buffer solution is phosphate buffer solution PBS. After the reaction at 4-80 ℃, the amount of ethylene glycol produced is detected by HPLC.
The reaction equation is shown in formula (2):
50mM methanol, 2mM anhydrous magnesium sulfate, 0.1mM TPP,25mM NADH,3mg methanol oxidase AOX,0.3mg catalase CAT,9mg glycolaldehyde synthase GALS,1mg ethanol dehydrogenase GOX0313 and 3mL total volume of the reaction system, 0.05M buffer solution, PBS with pH value of 7.0 and 35 ℃ are added into the reaction system, and after reaction for 8 hours, 0.89g/L ethylene glycol is detected. The reaction progress of the ethylene glycol production is shown in FIG. 3 (note: the abscissa indicates the calculated reaction time after the simultaneous addition of three enzymes).
Methanol oxidase AOX and catalase CAT were all purchased from Sigma. The glycolaldehyde synthase GALS gene was obtained from the literature (DOI No. 10.1038/s41467-019-09095 of this literature) based on which it was obtained by Huada gene synthesis. The alcohol dehydrogenase GOX0313 is obtained by a patent (patent number 202111284219.5) based on which it is obtained by Huada gene synthesis.
Example 3: catalytic production of D-erythrose by methanol oxidase, glycolaldehyde synthetase and D-erythrose synthetase three enzymes
The reaction system: the total volume of the reaction system is 3mL, and the buffer solution is phosphate buffer solution PBS. After the reaction at 4 to 80 ℃, the amount of D-erythrose produced was measured by HPLC.
The reaction equation is shown in formula (3):
the total volume of the reaction system was 3mL, the buffer was 0.05M, PBS with pH=9.0, 50mM methanol, 2mM anhydrous magnesium sulfate, 0.1mM TPP,25mM NAD +,3mg methanol oxidase AOX,0.3mg catalase CAT,6mg glycolaldehyde synthase GALS were added to react at 35℃for 8 hours, 2mg 2-deoxy-D-ribose 5-phosphate aldolase DERAeco protein was added, and after continuing the reaction at 65℃for 1 hour, 0.052g/L D-erythrose was detected. The D-erythrose yield reaction profile is shown in FIG. 4 (note: abscissa reaction time refers to the calculated reaction time after addition of DERAEco).
Methanol oxidase AOX and catalase CAT were all purchased from Sigma. The glycolaldehyde synthase GALS gene was obtained from the literature (DOI No. 10.1038/s41467-019-09095 of this literature) based on which it was obtained by Huada gene synthesis. The enzyme DERAEco is obtained from the patent (GenBank accession number: BAE78370.1 of the gene encoding the enzyme, patent number: 202111282875.1) (based on the patent, from Huada gene synthesis).
It should be noted that the above-described embodiments are only for explaining the present invention and do not constitute any limitation of the present invention. The invention has been described with reference to exemplary embodiments, but it is understood that the words which have been used are words of description and illustration, rather than words of limitation. Modifications may be made to the invention as defined in the appended claims, and the invention may be modified without departing from the scope and spirit of the invention. Although the invention is described herein with reference to particular means, materials and embodiments, the invention is not intended to be limited to the particulars disclosed herein, as the invention extends to all other means and applications which perform the same function.
Claims (1)
1. Multienzyme cascade catalysis CO 2 A method for synthesizing a 2C or 4C compound comprising a multienzyme cascade catalyzing CO 2 Synthesis of glycolic acid or multienzyme cascade catalytic CO 2 Synthesizing D-erythrose;
the multienzyme cascade catalyzes CO 2 The method for synthesizing the glycollic acid comprises the following steps:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) Glycolaldehyde is catalyzed by glycolaldehyde dehydrogenase to synthesize glycolaldehyde; wherein methanol catalyzes CO by photo, electro or multienzyme 2 Obtaining by transformation;
the methanol oxidase is AOX enzyme; the glycolaldehyde synthase is a GALS enzyme; the glycollic acid dehydrogenase is aldehyde dehydrogenase ALDH-BL21; the reaction system: substrates methanol, anhydrous magnesium sulfate, TPP, nad+, methanol oxidase AOX, catalase CAT, glycolaldehyde synthase GALS, glycolaldehyde dehydrogenase ALDH-BL21, genBank accession numbers of genes encoding glycolaldehyde dehydrogenase ALDH-BL 21: CAQ31929.1;
the multienzyme cascade catalyzes CO 2 The method for synthesizing glycollic acid comprises reacting substrate methanol, anhydrous magnesium sulfate, TPP, NAD+, methanol oxidase AOX, catalase CAT, glycolaldehyde synthetase GALS at 35 deg.C for 6h, adding acetaldehyde dehydrogenase ALDH-BL21, and reacting at 35 deg.C for 0.5 hr; the pH of the reaction was 8.0; the mass ratio of the methanol oxidase to the glycolaldehyde synthase to the glycolaldehyde dehydrogenase is 1:2:1;
the multienzyme cascade catalyzes CO 2 The method for synthesizing D-erythrose comprises the following steps:
(1) Methanol is used as a substrate, and methanol oxidase is utilized to convert the methanol into formaldehyde;
(2) Synthesizing glycolaldehyde by catalyzing formaldehyde with glycolaldehyde synthetase;
(3) Catalyzing glycolaldehyde to synthesize D-erythrose by using D-erythrose synthetase;
wherein methanol catalyzes CO by photo, electro or multienzyme 2 Obtaining by transformation;
the methanol oxidase is AOX enzyme; the glycolaldehyde synthase is a GALS enzyme; the D-erythrose synthetase is 2-deoxy-D-ribose 5-phosphate aldolase DERAEco;
the reaction system: substrate methanol, anhydrous magnesium sulfate, TPP, methanol oxidase AOX, catalase CAT, glycolaldehyde synthase GALS, D-erythrose synthase DERAEco, genBank accession numbers of genes encoding D-erythrose synthase DERAEco: BAE78370.1;
the multienzyme cascade catalyzes CO 2 The method for synthesizing D-erythrose comprises reacting substrate methanol, anhydrous magnesium sulfate, TPP, methanol oxidase AOX, catalase CAT and glycolaldehyde synthetase GALS at 35 deg.C for 8h, adding 2-deoxy-D-ribose 5-phosphate aldolase DERAeco, and reacting at 65deg.C for 1 hr; the pH of the reaction was 9; the ratio of methanol oxidase, glycolaldehyde synthase and D-erythrose synthase 3 enzymes was 1.5:3:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210306243.2A CN114807249B (en) | 2022-03-25 | 2022-03-25 | Catalytic CO 2 Multi-enzyme cascade pathway for synthesizing 2C or 4C compounds |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210306243.2A CN114807249B (en) | 2022-03-25 | 2022-03-25 | Catalytic CO 2 Multi-enzyme cascade pathway for synthesizing 2C or 4C compounds |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114807249A CN114807249A (en) | 2022-07-29 |
| CN114807249B true CN114807249B (en) | 2023-12-15 |
Family
ID=82530991
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210306243.2A Active CN114807249B (en) | 2022-03-25 | 2022-03-25 | Catalytic CO 2 Multi-enzyme cascade pathway for synthesizing 2C or 4C compounds |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114807249B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007228924A (en) * | 2006-03-02 | 2007-09-13 | Kaneka Corp | Method for producing glyoxylic acid |
| CN104017795A (en) * | 2014-05-27 | 2014-09-03 | 中国科学院天津工业生物技术研究所 | Biosynthesis method of 2-deoxy scarce aldose by using aldolase |
| CN110975938A (en) * | 2019-12-18 | 2020-04-10 | 华能国际电力股份有限公司 | Catalyst for preparing methanol by carbon dioxide hydrogenation and preparation method thereof |
| CN113755543A (en) * | 2020-08-24 | 2021-12-07 | 中国科学院天津工业生物技术研究所 | Starch biosynthesis method |
| CN114058601A (en) * | 2021-11-01 | 2022-02-18 | 北京化工大学 | Enzyme with function of catalyzing glycolaldehyde to synthesize ethylene glycol and application thereof |
-
2022
- 2022-03-25 CN CN202210306243.2A patent/CN114807249B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2007228924A (en) * | 2006-03-02 | 2007-09-13 | Kaneka Corp | Method for producing glyoxylic acid |
| CN104017795A (en) * | 2014-05-27 | 2014-09-03 | 中国科学院天津工业生物技术研究所 | Biosynthesis method of 2-deoxy scarce aldose by using aldolase |
| CN110975938A (en) * | 2019-12-18 | 2020-04-10 | 华能国际电力股份有限公司 | Catalyst for preparing methanol by carbon dioxide hydrogenation and preparation method thereof |
| CN113755543A (en) * | 2020-08-24 | 2021-12-07 | 中国科学院天津工业生物技术研究所 | Starch biosynthesis method |
| CN114058601A (en) * | 2021-11-01 | 2022-02-18 | 北京化工大学 | Enzyme with function of catalyzing glycolaldehyde to synthesize ethylene glycol and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| 2-deoxyribose-5-phosphate aldolase, NAD(P)-linked [Escherichia coli str. K-12 substr. W3110];Musso,R.;NCBI Database;第1-2页 * |
| gamma-aminobutyraldehyde dehydrogenase [Escherichia coli BL21(DE3)];Krempl,P.M;NCBI DATABASE;第1-2页 * |
| 氧化还原酶在多酶级联反应中的应用进展;应向贤;毛王伟;张丽;张洁;赵嫚;汪钊;;发酵科技通讯(03);全文 * |
| 草酸钙结石的研究进展;赵宇 等;医学研究生学报;第555-560页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114807249A (en) | 2022-07-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Pradima et al. | Review on enzymatic synthesis of value added products of glycerol, a by-product derived from biodiesel production | |
| KR101189818B1 (en) | Method for producing hydrocarbons and oxygen-containing compounds, from biomass | |
| EP1892300B1 (en) | Method for producing 1,3-propanediol using crude glycerol, a by-product from biodiesel production | |
| CN102712939A (en) | Process for the fermentative preparation of (3-hydroxy)propionaldehyde in the presence of semicarbazide or carbohydrazide | |
| WO2008124490A1 (en) | Methods of producing butanol | |
| AU2009344282A1 (en) | Process for the preparation of (3R)-hydroxybutyl (3R)-hydroxybutyrate by enzymatic enantioselective reduction employing Lactobacillus brevis alcohol dehydrogenase | |
| Held et al. | Biocatalysis. Biological systems for the production of chemicals | |
| JPWO2006126626A1 (en) | Method for producing glycolic acid | |
| Hata et al. | Enzymatic production of D-(-)-pantoyl lactone from ketopantoyl lactone | |
| FI81607C (en) | Preparation of butanol by improved fermentation process | |
| Ma et al. | C2 feedstock-based biomanufacturing of value-added chemicals | |
| CN114807249B (en) | Catalytic CO 2 Multi-enzyme cascade pathway for synthesizing 2C or 4C compounds | |
| Casy et al. | Enantioselective reduction of β, χ-unsaturated α-keto acids usingbacillus stearothermophilus lactate dehydrogenase: A new route to functionalised allylic alcohols | |
| CN114921392B (en) | Method for efficiently co-producing gluconic acid and allitol | |
| Oh et al. | Biohydrogen production from carbon monoxide and water by Rhodopseudomonas palustris P4 | |
| CN110423793B (en) | Biosynthesis method of 2-hydroxy-3-methyl-3-formylbutyric acid | |
| CN111170845B (en) | Method for producing lactic acid by catalytic conversion of glucose and application thereof | |
| EP2179050A1 (en) | Microbial kinetic resolution of ethyl-3,4-epoxybutyrate | |
| CN107663517A (en) | A kind of preparation method of L lactic acid catalystic converter system and L lactic acid | |
| CN101054597A (en) | Method for producing cystine by microorganism transformation | |
| Wagner et al. | Next-generation feedstocks methanol and ethylene glycol and their potential in industrial biotechnology | |
| CN106834366B (en) | Method for producing alpha-ketoglutaric acid by using L-glutamic acid dehydrogenase as catalyst | |
| Bornscheuer | (Chemo-) enzymatic cascade reactions | |
| CN116904382B (en) | Host cell and method for co-producing dihydroxyacetone and bose | |
| CN116355974B (en) | Production process for synthesizing furan ammonium salt by enzyme-chemical method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |