CN106755172B - New way for synthesizing acetyl coenzyme A and derivative products thereof by using glycolaldehyde - Google Patents

Abstract

The invention discloses a new way for synthesizing acetyl coenzyme A and derivative products thereof by using glycolaldehyde, which comprises a reaction of reacting glycolaldehyde with 3-glyceraldehyde phosphate to generate 5-arabinose phosphate under enzyme catalysis, wherein the enzyme is selected from aldolase, transaldolase, isozyme and mutase thereof. The method has the advantages of high catalytic rate, high reaction efficiency, low cost, strong affinity with substrates and high catalytic activity of auxiliary enzymes Rpi A and Rpe from a pentose phosphate pathway, high catalytic rate, carbon theoretical yield of a reaction route of 100 percent, no carbon loss, recycling of G3P, enzyme and coenzyme, and the like. The method of the preparation method can play a more obvious role in the production process of controllable substrate level, such as in-vitro continuous multi-enzyme catalysis, fed-batch fermentation, continuous fermentation and the like.

Description

A New Approach to Synthesize Acetyl-CoA and Its Derivatives Using Glycolaldehyde

technical field

The invention relates to the technical field of biomedicine, in particular to a method for synthesizing acetyl-CoA and derivatives thereof by using glycolaldehyde.

Background technique

Acetyl coenzyme A (AcCoA) is a key intermediate in the synthesis of essential biological compounds, including polyketides, fatty acids, isoprenoids, alkaloids, vitamins, and amino acids. Metabolites derived from AcCoA are primary and secondary metabolites, which include compounds of industrial utility. AcCoA can be generated from acetyl phosphate (AcP) (eg, catalyzed by phosphoacetyltransferase Pta (EC 2.3.1.8)), and then generate a series of biological products based on AcCoA, which are widely used in biocatalysis and other fields. The synthetic methods of AcP are as follows:

Patent application WO2015/144447A1 discloses a method utilizing phosphoketolase (EC 4.1.2.9, fructose-6-phosphate phosphoketolase EC4.1.2.22) or sulfoacetaldehyde acetyltransferase (EC 2.3.3.15) catalyzed Formaldehyde produces the method for acetylphosphoric acid, reaction equation is as follows:

2CH ₂ O+phosphoric acid→acetylphosphoric acid+H ₂ O.

The phosphoketolase used in the above preparation reaction has poor affinity with formaldehyde, the catalytic efficiency is less than 1% of the conversion of its optimum substrate 5-phosphate xylulose, and the tolerance of microbial cells to formaldehyde is generally poor. , the applicable expansion space is small.

Patent application WO2015/181074A1 discloses a method for catalyzing D-fructose to produce D-erythrose and acetyl phosphate using phosphoketolase (EC 4.1.2.9, fructose-6-phosphate phosphoketolase EC 4.1.2.22), which The method also includes converting D-erythrose into glycolaldehyde, which is further converted into acetyl phosphate, and the reaction process is as follows:

The reaction rate of the above-mentioned preparation method of acetyl phosphate is low, the conversion number K _cat of phosphoketolase to fructose is only 0.1/s, the efficiency is very low, and the reaction time is long (18h), which will raise the stability of the enzyme and the product. test. In addition, the third step of the cleavage of D-erythrose into glycolaldehyde is a reversible process, and its reaction rate is affected by the concentration of glycolaldehyde after the cleavage, making it difficult to accumulate high concentrations of glycolaldehyde in the system, which will lead to the formation of acetyl phosphate The rate is not guaranteed. The acetyl phosphate generated in the second enzymatic reaction is likely to affect the yield of the last enzymatic reaction due to product inhibition. If the acetyl phosphate generated in the second enzymatic reaction is removed immediately, the entire preparation process will be complicated.

SUMMARY OF THE INVENTION

In order to overcome the deficiencies of the prior art, one aspect of the present invention provides a method for preparing arabinose 5-phosphate (Ara5P), the method comprising the reaction of glycolaldehyde and glyceraldehyde 3-phosphate under enzyme catalysis to generate Ara5P, the The enzyme is selected from the group consisting of aldolase, transaldolase, and isoenzymes and mutant enzymes thereof.

Preferably, the reaction of the preparation is carried out by microorganisms;

Preferably, the aldolase is fructose bisphosphate aldolase (EC 4.1.2.13);

Preferably, the transaldolase is a mutant enzyme Tal B ^F178Y of the transaldolase (EC2.2.1.2) derived from Escherichia coli.

Another aspect of the present invention also provides a composition for preparing Ara5P, comprising:

(1) Glycoaldehyde,

(2) glyceraldehyde 3-phosphate,

(3) aldolase, transaldolase, or its isoenzyme, mutant enzyme;

or,

(1) Glycoaldehyde,

(2) glyceraldehyde 3-phosphate,

(3) Microorganisms expressing aldolase, transaldolase, or isozymes or mutant enzymes thereof;

Preferably, the aldolase is fructose bisphosphate aldolase (EC 4.1.2.13);

Preferably, the transaldolase is a mutant enzyme Tal B ^F178Y of the transaldolase (EC2.2.1.2) derived from Escherichia coli.

Another aspect of the present invention provides a preparation method of acetyl phosphate (AcP), the method comprises the steps of the above-mentioned preparation method of Ara5P; preferably, the preparation method of AcP further comprises the conversion of Ara5P into xylulose-5-phosphate ( Xu5P) is further converted into the step of AcP;

Preferably, the step of converting Ara5P into Xu5P comprises the step of converting Ara5P into ribulose 5-phosphate (Ru5P) and further converting it into Xu5P;

Preferably, the reaction of converting Xu5P into AcP comprises the step of generating AcP under the catalysis of phosphoketolase by Xu5P and phosphoric acid;

More preferably, the phosphoketolase is phosphoketolase (EC 4.1.2.9) or fructose-6-phosphate phosphoketolase (EC 4.1.2.22);

The phosphoketolase can be derived from the microorganism expression, artificial synthesis and purification process of various species, especially derived from microorganisms such as preferably derived from Pseudomonas stutzeri (such as Pseudomonas stutzeri Pseudomonas stutzeri A1501) and Bifidobacterium adolescentis strains. Phosphotransketolase can play a good catalytic role in the above reactions;

Preferably, the reaction system for converting Xu5P into AcP also includes a coenzyme of phosphoketolase, such as thiamine pyrophosphate;

Preferably, the reaction of the preparation is carried out by microorganisms.

Another aspect of the present invention provides a composition for preparing AcP, including the above-mentioned composition for preparing Ara5P;

Preferably, the above-mentioned composition for preparing AcP also includes phosphoric acid and phosphoketolase;

More preferably, the above-mentioned composition for preparing AcP also includes phosphoketolase coenzyme;

Preferably, the above-mentioned composition for preparing AcP also includes an enzyme that can catalyze Ara5P to generate Xu5P, the enzyme can be one kind of enzyme, or a combination of two or more kinds of enzymes, preferably ribose-5-phosphate isomerase in combination with ribulose-phosphate 3-isomerase;

In a preferred embodiment of the present invention, the above-mentioned composition for preparing AcP comprises:

(1) Glycoaldehyde,

(2) glyceraldehyde 3-phosphate,

(3) Aldolase, transaldolase, or its isoenzyme, mutant enzyme,

(4) ribose-5-phosphate isomerase,

(5) Ribulose-phosphate 3-isomerase,

(6) Phosphate, phosphoketolase, coenzyme.

Preferably, the aldolase is fructose bisphosphate aldolase (EC 4.1.2.13);

Preferably, the transaldolase is the mutant enzyme Tal B ^F178Y of the transaldolase (EC2.2.1.2) derived from Escherichia coli;

Preferably, the phosphoketolase is phosphoketolase (EC 4.1.2.9) or fructose-6-phosphate phosphoketolase (EC 4.1.2.22);

Preferably, the phosphoketolase coenzyme is thiamine pyrophosphate;

In a more preferred embodiment of the present invention, the above-mentioned composition for preparing AcP comprises:

(1) Glycoaldehyde,

(2) glyceraldehyde 3-phosphate,

(3) Fructose bisphosphate aldolase (EC 4.1.2.13) or a mutant enzyme Tal B ^F178Y of Escherichia coli-derived transaldolase (Tal B),

(4) ribose-5-phosphate isomerase (EC 5.3.1.6),

(5) Ribulose-phosphate 3-isomerase (EC 5.1.3.1),

(6) Phosphate, phosphoketolase (EC 4.1.2.9) or fructose-6-phosphate phosphoketolase (EC 4.1.2.22) and thiamine pyrophosphate.

The stability of AcP is poor, and it is often made into the form of acetyl phosphate in practical applications. On the other hand, the present invention provides a preparation method of acetyl phosphate, which comprises the above-mentioned steps of the preparation method of Ara5P;

Preferably, the preparation method of the acetyl phosphate salt comprises the steps of the above-mentioned preparation method of AcP;

Preferably, the acetyl phosphate includes: acetyl phosphate dilithium salt, acetyl phosphate disodium salt, acetyl phosphate diammonium salt.

Another aspect of the present invention provides a method for preparing acetyl coenzyme A (AcCoA), the method comprising the steps of the above-mentioned preparation method of Ara5P; preferably, the above-mentioned preparation method of AcCoA comprises the steps of the above-mentioned preparation method of AcP;

Preferably, the preparation method of AcCoA further comprises the step of converting AcP into AcCoA under the action of acetyl phosphotransferase (Pta);

Preferably, the reaction of the preparation is carried out by microorganisms.

Another aspect of the present invention provides a method for preparing an AcCoA derivative compound, the method comprising the steps of the above-mentioned preparation method of Ara5P, preferably, the above-mentioned steps of the preparation method of AcP, more preferably, the above-mentioned steps of the preparation method of AcCoA; in the present invention In a preferred embodiment, the AcCoA derivative compound is selected from: acetone, isopropanol, acetic acid, L-glutamic acid, L-glutamine, L-proline, L-arginine, L- Leucine, L-cysteine, succinate and polyhydroxyalkanoate;

In another preferred embodiment of the present invention, the AcCoA derivative compound is poly-3-hydroxybutyrate (PHB);

Preferably, the reaction of the preparation is carried out by microorganisms.

The glyceraldehyde-3-phosphate described in the preparation method of the above-mentioned Ara5P, AcP, acetyl phosphate, AcCoA, AcCoA derivative compounds can be purchased commercially, and can also be prepared by methods known in the prior art, such as using glucose, glycerol , dihydroxyacetone phosphate, etc., for example, dihydroxyacetone phosphate can be converted into glyceraldehyde 3-phosphate under the action of triose phosphate isomerase; also can be prepared by adding glyceraldehyde 3-phosphate in the reaction system of the above preparation method The reactants are prepared online; the source of glyceraldehyde-3-phosphate does not limit the scope of the present invention.

Glycolaldehyde described in the preparation method of above-mentioned Ara5P, AcP, acetyl phosphate, AcCoA, AcCoA derivative compound can be prepared by methods known in the prior art, such as, the halogenation reaction of acetaldehyde, the cleavage reaction of carbohydrates, etc. , wherein, the cost that utilizes formaldehyde to prepare glycolaldehyde is lower, preferably, above-mentioned glycolaldehyde is to utilize formaldehyde according to prior art preferably as the preparation method and the technique of the glycolaldehyde recorded in " formaldehyde synthesizes glycolaldehyde research and its application progress ". Xin Kun, Li Qingsong, Jia Bing, et al. Research progress on the synthesis of glycolaldehyde from formaldehyde and its application. Natural gas chemical industry·C1 chemistry and chemical industry, 2016(41): 88-94) Prepared.

Another aspect of the present invention provides the use of the above-mentioned aldolase, transaldolase or its isoenzyme and mutant enzyme in the preparation of Ara5P, AcP, acetylphosphate, AcCoA, and AcCoA derivative compounds.

Another aspect of the present invention also provides the use of a microorganism expressing the above-mentioned aldolase, transaldolase or its isoenzyme and mutant enzyme in the preparation of Ara5P, AcP, acetylphosphate, AcCoA, and AcCoA derivative compounds.

The enzymes involved in the present invention can be derived from various microbial sources, isoenzymes and mutant enzymes obtained by artificial transformation.

The reaction of the above preparation is carried out by microorganisms, such as one or more than two of the reactants involved in the reaction, and/or one or more than two kinds of enzymes can be produced online by microorganisms.

In the preparation method of Ara5P provided by the present invention, aldolase or transaldolase does not depend on coenzyme, the catalytic efficiency is high (V _max is greater than 50U/mg), and there are many kinds of aldolase and transaldolase, which can be optimized Lots of space. The catalytic substrate of phosphoketolase in the present invention is xylulose 5-phosphate, which is the most suitable substrate, with high affinity and enzyme activity (K _m is about 10-45 mM, V _max is 90-800 U/mg) , and as the last step of the reaction, the catalytic process is irreversible, so that the system is not limited by the reaction equilibrium, which can not only drive the rate of aldolase and transaldolase, but also maximize the yield of acetyl phosphate and promote 3- Replenishment of glyceraldehyde phosphate. The preparation method of AcP provided has a high catalytic rate, the theoretical carbon yield of the reaction route is 100%, there is no carbon loss, G3P and enzymes can be recycled, the reaction efficiency is high, and the cost is reduced. The auxiliary enzymes RpiA and Rpe of the sugar pathway have strong affinity with the substrate, good stability and high catalytic activity. The method of the above preparation method will exert more obvious advantages in the production process of controlled substrate level, such as fed-feed fermentation and continuous fermentation.

Description of drawings

Figure 1 shows a schematic diagram of the biosynthetic pathway provided in Example 1 of the present invention.

Figure 2 shows the GC-MS detection results provided in Example 6 of the present invention.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

The biosynthesis of acetyl phosphate using glycolaldehyde involved in the specific embodiment of the present invention is shown in FIG. 1 , and the information on the reactants, intermediates and enzymes involved in the reaction process is shown in Tables 1 and 2.

Table 1 Metabolite information table involved in the present invention

Table 2 Enzyme information table involved in the present invention

Example 1

Under the initial concentration of glycolaldehyde of 15mM, under the action of the mutant enzyme Tal B ^F178Y of Transaldolase Tal B derived from Escherichia coli (the 178th amino acid residue was mutated from F to Y on the basis of the original enzyme), 5mM G3P was added to carry out. The preparation reaction of Ara5, after 1 h of reaction, the reaction solution was lyophilized and derivatized, the content of Ara5P quantitatively detected by GC-MS was 2.5mM, and the average synthesis rate of Ara5P was 52.1 μmol (Ara5P)/min/mg (enzyme protein).

Example 2

At the initial concentration of 15 mM, Glycolaldehyde, ThDP concentration of 1 mM, PO ₄ ³⁺ concentration of 2 mM, Bifidobacterium adolescentis derived Fpk (EC 4.1.2.22), Escherichia coli derived aldolase (Fsa, EC 4.1.2.13), 2 mM were added G3P and a small amount of Rpi A and Rpe were used to prepare AcP. After testing, the conversion rate of glycolaldehyde was 16.3 μmol (glycolaldehyde)/min/mg (enzyme protein).

The above data is the average rate of the reaction system for 3h.

Example 3

Glycoaldehyde was added at an initial concentration of 15 mM, ThDP at 1 mM and PO ₄ ³⁺ at 2 mM, adding Xpk (EC 4.1.2.9) derived from Pseudomonas stutzeri A1501, Transaldolase (EC 4.1.2.9) derived from Escherichia coli Tal B, EC 2.2.1.2) mutant enzyme Tal B ^F178Y , 2 mM G3P and trace amounts of Rpi A and Rpe, were used to prepare AcP. After testing, the conversion rate of glycolaldehyde was 19.7 μmol (glycolaldehyde)/min/mg (enzyme protein).

The above data is the average rate of the reaction system for 3h.

Example 4

Glycoaldehyde at an initial concentration of 20 mM, ThDP concentration of 1 mM, PO ₄ ³⁺ concentration of 2 mM, added Xpk derived from Pseudomonas stutzeri A1501, Transaldolase derived from Escherichia coli (tal B, EC 2.2.1.2 ) of the mutant enzyme Tal B ^F178Y , 4 mM DHAP and a small amount of Rpi A, Rpe, TpiA, to carry out the preparation reaction of AcP. After testing, the conversion rate of glycolaldehyde was 26.5 μmol (glycolaldehyde)/min/mg (enzyme protein).

The above data is the average rate of the reaction system for 3h.

Example 5

The genes for the synthesis of phosphoketolase were integrated into the genome of Escherichia coli K-12 MG1655, and then the genes for the synthesis of poly-3-hydroxybutyrate (PHB) (PhaA, PhaB and PhaC) and TalB ^F178Y were incorporated The plasmid of the gene is transferred into the above strain, and the above plasmid selects the IPTG inducible promoter (the rest of the required enzymes are already present in E. coli). The above strains were cultured in LB medium. After culturing for 2.5 hours, when the OD ₆₀₀ value of the bacteria reached 0.8-1.0, IPTG was added to a final concentration of 0.5mM to induce (22°C, 6h) to express the target enzyme protein on the above plasmid . After centrifugation (4°C, 8000r/min, 5min) to collect the cells, use the M9 medium without carbon and nitrogen sources to suspend the cells, and then re-centrifuge to collect the cells. This suspension and centrifugation process is repeated three times to remove the cells. Residual carbon source in bacteria. After the final collected cells were resuspended in the above-mentioned M9 medium, they were equally divided into 3 parts, and 0.02g, 0.45g and 0.9g of glycolaldehyde were added respectively (the cells contained the intermediate metabolite G3P, no additional addition was required), The above-mentioned M9 medium was uniformly adjusted to 30 mL, fermented for 20 h, and the cell growth during the fermentation was observed. After the fermentation, the cells were collected and the PHB content in the fermentation broth was detected.

Detection of PHB: After centrifuging the fermentation broth to obtain bacterial cells, freeze-dried and weighed. Use 4 mL of a mixture of chloroform and esterification solution (the main component is methanol) with a volume ratio of 1:1 to derivatize the bacterial cell freeze-dried powder at 100 °C for 4 hours, then add 2 mL of ultrapure water, stand for stratification, remove methanol and Cell debris, etc., the lower layer of methyl 3-hydroxybutyrate chloroform solution was removed, and the methyl 3-hydroxybutyrate in chloroform was quantitatively detected by GC-MS.

Experimental results: During the above fermentation process, the bacteria can grow and reproduce normally; after the fermentation, the corresponding dry weights of the bacteria are 0.025g, 0.076g, and 0.131g respectively; after comparing with the standard product, the samples were measured. The total content of 3-hydroxybutyric acid was 0.005g, 0.066g and 0.089g respectively, and the conversion rate of carbon source was between 10% and 20%. The results show that the bacteria can use glycolaldehyde and G3P and enzymes contained in it to synthesize acetyl phosphate and acetyl-CoA, and continue to generate PHB, a derivative of acetyl-CoA, for its own carbon source storage. It does not show obvious cytotoxicity and can maintain the normal metabolic consumption of the bacteria.

Example 6

Acetyl phosphate and AcCoA were prepared in different ways, and reacted for 2 h. After the reaction, the contents of Ara5P and AcCoA were detected. The content of AcCoA can also indirectly represent the synthetic amount of acetyl phosphate. The reactants, enzymes and their additions are shown in Table 3.

Detection of Ara5P: Take 50 μL of the reaction solution, freeze-dry the reaction solution, and derivatize it with 30 μL of methoxyamine hydrochloride and 90 μL of trimethylsilyl trifluoroacetamide for 1 h, respectively, at a derivatization temperature of 37 °C. Detection of Ara5P content.

Detection of AcCoA: Take 50 μL of the reaction solution, stop the reaction with 10% sulfuric acid solution, remove impurities with a 0.22 μm filter membrane, and perform liquid phase detection. Mobile phase A is 0.2 M sodium dihydrogen phosphate solution with pH=5 , the mobile phase B was acetonitrile, and the maximum absorption peak of AcCoA appeared at 254 nm.

The product content detection results are shown in Table 3.

The results in Table 3 show that when aldolase or transaldolase is not added to the above reaction system, no Ara5P is generated, the amount of AcCoA produced is also very small, and the reaction conversion rate is very low; adding aldolase or transaldolase to the above reaction system After alcoholase and G3P, Ara5P can be generated, and the reaction conversion rate is high; after adding RpiA and Rpe at the same time, the generated Ara5P will be converted into acetyl phosphate under the action of RpiA, Rpe and F/Xpk, and further by phosphate acetyltransferase It is converted into AcCoA, and the detection results of Ara5P and AcCoA show that the reaction conversion rate is very high; adding DHAP and TpiA to the system at the same time, instead of G3P, can also obtain similar effects as directly adding G3P, and achieve a higher conversion rate.

Table 3 Substance addition and metabolite generation in the system

(unit of concentration of reactant or product: mM; unit of enzyme dosage: μg)

Remarks: The buffer contains three main components, Tris, NaCl and MgCl ₂ , pH7.5, 37℃.

In addition, in the same reaction system containing reactants such as GA, Phos, G3P, ThDP, etc., respectively: A: without adding aldolase or transaldolase; B: adding Tal B ^F178Y ; C: adding Tal B ^F178Y and F/Xpk; D: Add Tal B ^F178Y , F/Xpk and Rpi A; E: Add Tal B ^F178Y , F/Xpk and Rpi A and Rpe. The content of Ara5P was detected by GC-MS, and the results were shown in the AE diagram in Figure 2 (the peak time of Ara5P was 37 min). It can be seen from the graphs in Fig. 2 and the corresponding reaction conditions that Ara5P is generated only after the addition of aldolase; only when Rpi A and Rpe are added at the same time, the generated Ara5P will be completely destroyed by phosphoketolase. break down.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention. within.

Claims (26)

1. A method for preparing 5-arabinose phosphate, which comprises reacting glycolaldehyde with 3-glyceraldehyde phosphate under the catalysis of an enzyme to produce the 5-arabinose phosphate, wherein the enzyme is selected from the group consisting of: aldolase, transaldolase, isozymes thereof, and mutant enzymes thereof.

2. The method of claim 1, wherein the reaction of preparation is carried out by a microorganism.

3. The method of claim 1, wherein the aldolase is fructose bisphosphate aldolase (EC4.1.2.13); and/or the presence of a gas in the gas,

the transaldolase is mutant enzyme TalB of transaldolase (EC2.2.1.2) derived from Escherichia coli^F178Y。

4. The process according to any one of claims 1 to 3, wherein the glycolaldehyde is prepared from formaldehyde.

5. The method of any one of claims 1-3, wherein the glyceraldehyde-3-phosphate is prepared from dihydroxyacetone phosphate.

6. A composition for preparing arabinose 5-phosphate comprising:

(1) the reaction product of the alcohol aldehyde and the alcohol aldehyde,

(2) the 3-phosphoric acid of the glyceraldehyde is taken as a raw material,

(3) aldolase, transaldolase, or isozyme or mutant enzyme thereof;

or the like, or, alternatively,

(1) the reaction product of the alcohol aldehyde and the alcohol aldehyde,

(2) the 3-phosphoric acid of the glyceraldehyde is taken as a raw material,

(3) a microorganism expressing an aldolase, transaldolase, or an isozyme or a mutant enzyme thereof.

7. The composition of claim 6, wherein said aldolase is fructose bisphosphate aldolase (EC4.1.2.13); and/or the presence of a gas in the gas,

the transaldolase is mutant enzyme TalB of transaldolase derived from Escherichia coli^F178Y。

8. A method for producing acetyl phosphate, which comprises the method for producing arabinose 5-phosphate according to any one of claims 1 to 5.

9. The method of claim 8, wherein the reaction of preparation is carried out by a microorganism.

10. The method of claim 8, further comprising the step of converting arabinose 5-phosphate to xylulose 5-phosphate and further to acetyl phosphate.

11. The method of claim 10, wherein the step of converting arabinose 5-phosphate to xylulose 5-phosphate comprises the step of converting arabinose 5-phosphate to ribulose 5-phosphate and further to xylulose 5-phosphate; and/or the presence of a gas in the gas,

the reaction for converting xylulose 5-phosphate into acetyl phosphate comprises the step of generating acetyl phosphate by xylulose 5-phosphate and phosphoric acid under the catalysis of phosphoketolase.

12. The method of claim 11, wherein said phosphoketolase is phosphoketolase (EC4.1.2.9) or fructose-6-phosphate phosphoketolase (EC4.1.2.22).

13. A composition for preparing acetyl phosphate comprising the composition for preparing arabinose 5-phosphate according to claim 6.

14. The composition of claim 13, wherein the composition further comprises a phosphate, phosphoketolase.

15. The composition of claim 14, wherein the composition further comprises a phosphoketolase coenzyme; and/or the presence of a gas in the gas,

the composition also includes an enzyme that catalyzes the production of xylulose-5-phosphate from arabinose-5-phosphate.

16. The composition of claim 15, wherein the composition comprises:

(1) the reaction product of the alcohol aldehyde and the alcohol aldehyde,

(2) the 3-phosphoric acid of the glyceraldehyde is taken as a raw material,

(3) aldolase, transaldolase, or isozymes, mutant enzymes thereof,

(4) a ribose-5-phosphate isomerase enzyme,

(5) a ribulose-phosphate 3-isomerase,

(6) phosphoric acid, phosphoketolase and coenzyme.

17. The composition of claim 16, wherein said aldolase is fructose bisphosphate aldolase (EC4.1.2.13); and/or the presence of a gas in the gas,

the transaldolase is mutant enzyme TalB of transaldolase derived from Escherichia coli^F178Y(ii) a And/or the presence of a gas in the gas,

the phosphoketolase is phosphoketolase (EC4.1.2.9) or fructose-6-phosphoketolase (EC4.1.2.22).

18. A method for preparing an acetyl phosphate, the method comprising the method for preparing arabinose 5-phosphate according to any one of claims 1 to 5 or the method for preparing acetyl phosphate according to any one of claims 8 to 12.

19. The method of claim 18, wherein said acetyl phosphate comprises: lithium acetyl phosphate, disodium acetyl phosphate and diammonium acetyl phosphate.

20. A method for producing acetyl-coa, which comprises the method for producing arabinose 5-phosphate according to any one of claims 1 to 5 or the method for producing acetyl phosphate according to any one of claims 8 to 12.

21. The method of claim 20, wherein the reaction of preparation is carried out by a microorganism.

22. A method for producing an acetyl-coa derivative compound, which comprises the method for producing arabinose 5-phosphate according to any one of claims 1 to 5 or the method for producing acetyl phosphate according to any one of claims 8 to 12 or the method for producing acetyl-coa according to claim 20 or 21.

23. The method of claim 22, wherein said acetyl-CoA derivative compound is selected from the group consisting of acetone, isopropanol, acetic acid, L-glutamic acid, L-glutamine, L-proline, L-arginine, L-leucine, L-cysteine, succinate, and polyhydroxyalkanoates.

24. The method of claim 22, wherein said acetyl-coa derived compound is poly-3-hydroxybutyrate.

25. The method of claim 22, wherein the reaction of preparation is carried out by a microorganism.

26. Use of an aldolase, transaldolase or an isozyme or a mutant thereof, or a microorganism expressing an aldolase, transaldolase or an isozyme or a mutant thereof, for the preparation of arabinose-5-phosphate, acetyl-coa derived compounds.

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