CN106755172B

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

Info

Publication number: CN106755172B
Application number: CN201611167145.6A
Authority: CN
Inventors: 马红武; 杨雪; 袁倩倩; 江会锋
Original assignee: Tianjin Institute of Industrial Biotechnology of CAS
Current assignee: Tianjin Institute of Industrial Biotechnology of CAS
Priority date: 2016-12-16
Filing date: 2016-12-16
Publication date: 2020-07-28
Anticipated expiration: 2036-12-16
Also published as: WO2018107880A1; CN106755172A

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

New way for synthesizing acetyl coenzyme A and derivative products thereof by using glycolaldehyde

Technical Field

The invention relates to the technical field of biomedicine, in particular to a method for synthesizing acetyl coenzyme A and derivative products thereof by using glycolaldehyde.

Background

Acetyl-coa (acacoa) is a key intermediate in the synthesis of essential biological compounds including polyketides, fatty acids, isoprenoids, alkaloids, vitamins, and amino acids, among others. Metabolites derived from acaoa are primary and secondary metabolites, which include industrially useful compounds. AcCoA can be generated from acetyl phosphate (AcP) (for example, the AcCoA can be obtained by catalyzing phosphate acetyltransferase Pta (EC 2.3.1.8)), and then a series of biological products taking the AcCoA as a platform are generated, so that the AcCoA can be widely applied to the fields of biological catalysis and the like. The synthesis method of AcP comprises the following steps:

patent application WO2015/144447a1 discloses a method for catalyzing formaldehyde to produce acetyl phosphate using phosphoketolase (EC4.1.2.9, fructose-6-phosphate phosphoketolase EC4.1.2.22) or sulfoacetaldehyde acetyltransferase (EC 2.3.3.15), the reaction equation is as follows:

2CH₂o + phosphoric acid → acetyl phosphoric acid + H₂O。

The phosphoketolase used in the preparation reaction has poor affinity with formaldehyde, catalytic efficiency is not as high as one percent of that of the phosphoketolase used for converting the optimal substrate 5-xylulose phosphate, and microbial cells have generally poor tolerance to formaldehyde and have small application and expansion space.

Patent application WO2015/181074a1 discloses a method for producing D-erythrose and acetyl phosphate by catalyzing D-fructose with phosphoketolase (EC4.1.2.9, fructose-6-phosphate phosphoketolase EC4.1.2.22), which further comprises converting D-erythrose into glycolaldehyde and then further into acetyl phosphate, the reaction process is as follows:

the reaction rate of the preparation method of the acetyl phosphate is low, and the transformation number K of phosphoketolase to fructose is_catOnly 0.1/s, very low efficiency and long reaction times (18h) were challenging for enzyme and product stability. In addition, the third stepThe reaction for cracking D-erythrose into glycolaldehyde is a reversible process, and the reaction rate is influenced by the concentration of the glycolaldehyde after cracking, so that high-concentration glycolaldehyde is difficult to accumulate in a system, and the generation rate of acetyl phosphate is difficult to guarantee. The acetyl phosphate formed by the second enzymatic reaction is likely to affect the yield of the final enzymatic reaction due to product inhibition, and if the acetyl phosphate formed by the second enzymatic reaction is removed immediately after the formation of acetyl phosphate, the operation of the whole production process is complicated.

Disclosure of Invention

In order to overcome the disadvantages of the prior art, the present invention provides, in one aspect, a method for preparing arabinose 5-phosphate (Ara5P), the method comprising reacting glycolaldehyde and glyceraldehyde 3-phosphate under an enzyme catalysis to produce Ara5P, the enzyme being selected from the group consisting of: aldolase, transaldolase, isozymes thereof, and mutant enzymes thereof.

Preferably, the reaction of the preparation is carried out by a microorganism;

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

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

In another aspect, the present invention provides a composition for preparing Ara5P, 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;

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

In another aspect, the present invention provides a process for the preparation of acetyl phosphate (AcP), said process comprising the process steps of the above preparation of Ara 5P; preferably, the preparation method of AcP further comprises the step of converting Ara5P into xylulose 5-phosphate (Xu5P) and further converting the xylulose 5-phosphate into AcP;

preferably, said step of converting Ara5P into Xu5P comprises the step of converting Ara5P into ribulose-5-phosphate (Ru5P) and further converting into Xu 5P;

preferably, the reaction for converting Xu5P into AcP comprises the steps of generating AcP by Xu5P and phosphoric acid under the catalysis of phosphoketolase;

more preferably, the phosphoketolase is phosphoketolase (EC4.1.2.9) or fructose-6-phosphate phosphoketolase (EC 4.1.2.22);

the phosphoketolase can be obtained from microbial expression, artificial synthesis and purification processes of various species, and particularly, the phosphoketolase derived from microorganisms such as Pseudomonas stutzeri (such as Pseudomonas stutzeri A1501) and Bifidobacterium adolescentis strains can play a good catalytic role in the reaction;

preferably, the reaction system for converting Xu5P into AcP further comprises a coenzyme of phosphoketolase, such as thiamine pyrophosphate;

preferably, the reaction of the preparation is carried out by a microorganism.

In another aspect, the present invention provides a composition for preparing AcP, comprising the above composition for preparing Ara 5P;

preferably, the composition for preparing AcP further comprises phosphoric acid, phosphoketolase;

more preferably, the composition for preparing AcP further comprises a phosphoketolase coenzyme;

preferably, the composition for preparing AcP further comprises an enzyme capable of catalyzing Ara5P to generate Xu5P, wherein the enzyme can be one enzyme or a combination of more than two enzymes, and is preferably a combination of ribose-5-phosphate isomerase and ribulose-phosphate 3-isomerase;

in a preferred embodiment of the present invention, the above composition for preparing AcP 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.

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

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

Preferably, the phosphoketolase is phosphoketolase (EC4.1.2.9) 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 composition for preparing AcP 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) fructose bisphosphate aldolase (EC4.1.2.13) or mutant enzyme of Escherichia coli-derived transaldolase (Tal B)^F178Y，

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

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

(6) phosphoketolase, phosphoketolase (EC4.1.2.9) or fructose-6-phosphate phosphoketolase (EC4.1.2.22), and thiamine pyrophosphate.

AcP has poor stability and is often prepared into an acetyl phosphate form in practical application, and the invention provides a preparation method of the acetyl phosphate in another aspect, wherein the preparation method comprises the steps of the Ara5P preparation method;

preferably, the preparation method of the acetyl phosphate comprises the preparation method steps of the AcP;

preferably, the acetyl phosphate comprises: lithium acetyl phosphate, disodium acetyl phosphate and diammonium acetyl phosphate.

In another aspect, the present invention provides a method for preparing acetyl-coenzyme a (acaco), which comprises the steps of the above preparation method of Ara 5P; preferably, the preparation method of AcCoA comprises the steps of the preparation method of AcP;

preferably, the preparation method of AcCoA further comprises the step of converting AcP into AcCoA under the action of acetylphosphotransferase (Pta);

preferably, the reaction of the preparation is carried out by a microorganism.

In another aspect, the present invention provides a method for preparing an AcCoA derivative compound, comprising the steps of the above preparation method of Ara5P, preferably, the above preparation method of AcP, more preferably, the above preparation method of AcCoA, wherein in a preferred embodiment of the present invention, the AcCoA 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 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 a microorganism.

Glyceraldehyde 3-phosphate described in the above preparation method of Ara5P, AcP, acetyl phosphate, aca a derivative compound may be commercially available, or may be prepared by a method known in the art, such as preparation using glucose, glycerol, dihydroxyacetone phosphate, etc., for example dihydroxyacetone phosphate may be converted to glyceraldehyde 3-phosphate by triose phosphate isomerase; the reactant prepared from 3-glyceraldehyde phosphate can be added into the reaction system of the preparation method to realize on-line preparation; the source of glyceraldehyde 3-phosphate does not limit the scope of the present invention.

The glycolaldehyde in the preparation method of Ara5P, acap, acetyl phosphate, acacoa, and acacoa derivative compounds can be prepared by methods known in the art, such as acetaldehyde halogenation, saccharide cleavage, etc., wherein the cost of preparing glycolaldehyde from formaldehyde is low, preferably, the glycolaldehyde is prepared by using formaldehyde according to the prior art, preferably, the preparation method and process of glycolaldehyde described in "research on synthesis of glycolaldehyde from formaldehyde and progress of its application" (xinkun, laiqingsong, jiabing, etc.. research on synthesis of glycolaldehyde from formaldehyde and progress of its application, chemical industry of natural gas, C1, 2016(41), (88-94).

The invention also provides application of the aldolase, the transaldolase or the isozyme and the mutant enzyme thereof in preparing Ara5P, AcP, acetyl phosphate, AcCoA and AcCoA derivative compounds.

The invention also provides application of the microorganism expressing the aldolase, the transaldolase or the isozyme and the mutant enzyme thereof in preparing Ara5P, AcP, acetyl phosphate, AcCoA and AcCoA derivative compounds.

The enzyme related to the invention can be from various microbial sources and artificially modified isoenzymes and mutant enzymes.

The reaction of the above preparation is carried out by a microorganism, for example, one or more than two of the reactants involved in the reaction, and/or one or more than two of the enzymes may be produced on-line by a microorganism.

In the preparation method of Ara5P, aldolase or transaldolase does not depend on coenzyme, and the catalytic efficiency is high (V)_maxMore than 50U/mg), and the aldolase and transaldolase are various in variety, and the optimization space is large. The catalytic substrate of the phosphoketolase is 5-xylulose phosphate, which is the most suitable substrate, and the affinity and the enzyme activity are high (K)_mAbout 10-45mM, V_max90-800U/mg) and as the last step of reaction, the catalytic process is irreversible, so that the system is not limited by reaction balance, the rates of aldolase and transaldolase can be pulled, the yield of acetyl phosphate can be improved to the maximum extent, and the anaplerosis of 3-phosphoglyceraldehyde can be promoted. The provided preparation method of AcP has higher catalytic rate, and the theoretical yield of carbon in the reaction route is 100 percentThe method has the advantages of no carbon loss, cyclic use of G3P and enzyme, high reaction efficiency, reduced cost, strong affinity with substrates, good stability and high catalytic activity of auxiliary enzymes RpiA and Rpe from the pentose phosphate pathway. The method of the preparation method can play a more obvious advantage in the production process of feeding fermentation, continuous fermentation and the like which can control the substrate level.

Drawings

FIG. 1 is a schematic diagram of a biosynthetic pathway provided in example 1 of the present invention.

FIG. 2 shows the GC-MS detection results provided in example 6 of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The biosynthetic pathway of acetyl phosphate using glycolaldehyde according to the embodiment of the present invention is shown in fig. 1, and the information of the reactants, intermediates and enzymes involved in the reaction process is detailed in tables 1 and 2.

TABLE 1 metabolite information Table referred to in the present invention

TABLE 2 enzyme information Table referred to in the present invention

Example 1

Initial ethanol aldehyde concentration of 15mMAt the initial concentration, the mutant enzyme, Tal B, of Transaldolase Tal B derived from Escherichia coli^F178Y(the 178 th amino acid residue was mutated from F to Y based on the proenzyme) 5mM G3P was added to carry out the preparation reaction of Ara5, after 1 hour of reaction, the reaction solution was lyophilized and derivatized, and the Ara5P content was 2.5mM and the average synthesis rate of Ara5P was 52.1. mu. mol (Ara5P)/min/mg (enzyme protein) as quantitatively determined by GC-MS.

Example 2

Glycolaldehyde at an initial concentration of 15mM, ThDP concentration of 1mM, PO₄ ³⁺The AcP preparation reaction was carried out at a concentration of 2mM by adding Fpk derived from Bifidobacterium adolescentis (EC4.1.2.22), aldolase derived from Escherichia coli (Fsa, EC4.1.2.13), 2mM G3P, and trace amounts of Rpi A and Rpi. The conversion rate of glycolaldehyde was determined to be 16.3. mu. mol (glycolaldehyde)/min/mg (enzyme protein).

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

Example 3

Glycolaldehyde at an initial concentration of 15mM, ThDP concentration of 1mM, PO₄ ³⁺The mutant enzyme Tal B derived from Xpk (EC4.1.2.9) derived from Pseudomonas stutzeri A1501 and Transaldolase (Tal B, EC2.2.1.2) derived from Escherichia coli were added to the mixture at a concentration of 2mM^F178Y2mM G3P and trace amounts of Rpi A and Rpe. The conversion rate of glycolaldehyde was determined to be 19.7. mu. mol (glycolaldehyde)/min/mg (enzyme protein).

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

Example 4

Glycolaldehyde at an initial concentration of 20mM, ThDP concentration of 1mM, PO₄ ³⁺The 2mM concentration was added to Xpk derived from Pseudomonas stutzeri A1501 and to talB, a mutant enzyme of Transaldolase (Tal B, EC2.2.1.2) derived from Escherichia coli^F178Y4mM DHAP and a small amount of Rpi A, Rpe, TpiA. The conversion rate of glycolaldehyde was determined to be 26.5. mu. mol (glycolaldehyde)/min/mg (enzyme protein).

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

Example 5

The gene for synthesizing phosphoketolase was integrated into the genome of Escherichia coli K-12 MG1655, and then the gene (PhaA, PhaB and PhaC) for synthesizing poly-3-hydroxybutyrate (PHB) and TalB were incorporated^F178YThe plasmid of (2) was transformed into the above strain, the above plasmid selected for the I PTG inducible promoter (the remaining required enzymes, all present in E.coli.) the above strain was cultured in L B medium for 2.5h, after which the OD of the cells was obtained₆₀₀When the value reaches 0.8-1.0, IPTG is added to the final concentration of 0.5mM, target enzyme protein on the plasmid is induced (22 ℃, 6h) to express, after the thalli is collected by centrifugation (4 ℃, 8000r/min, 5min), M9 culture medium without carbon source and nitrogen source is used for suspending the thalli, the thalli is collected by recentrifugation, the suspending and centrifuging process is repeated for three times for removing residual carbon source in the thalli, after the thalli collected finally is resuspended by the M9 culture medium, 3 parts are divided equally, 0.02G, 0.45G and 0.9G of glycolaldehyde (the thalli contains intermediate metabolite G3P and does not need to be added additionally) are respectively added, the volume is uniformly determined to 30M L by the M9 culture medium, the fermentation is carried out for 20h, the cell growth condition in the fermentation process is observed, and the thalli content of PHB in the fermentation liquid is collected and detected after the fermentation is finished.

And (3) detecting PHB, namely centrifuging the fermentation liquor to obtain thalli, freeze-drying and weighing, deriving the thalli freeze-dried powder for 4 hours at 100 ℃ by using a chloroform and esterification liquid (methanol is a main component) mixed solution 4m L with a volume ratio of 1:1, then adding 2m L ultrapure water, standing and layering, removing methanol, cell debris and the like, taking a lower-layer chloroform solution of 3-hydroxybutyrate methyl ester, and quantitatively detecting the 3-hydroxybutyrate methyl ester in chloroform by using a gas chromatograph-mass spectrometer GC-MS.

The experimental results are as follows: in the fermentation process, the thallus can grow and reproduce normally; after fermentation, the dry weight of the corresponding thallus is 0.025g, 0.076g and 0.131g respectively; comparing with a standard product, the total content of the 3-hydroxybutyric acid in the sample is respectively 0.005g, 0.066g and 0.089g, and the carbon source conversion rate is between 10 and 20 percent. The results show that the thalli can utilize glycolaldehyde, G3P and enzyme contained in the thalli to synthesize acetyl phosphate and acetyl coenzyme A, and continuously generate PHB which is a derivative of acetyl coenzyme A and is used for storing a carbon source of the thalli, the glycolaldehyde does not show obvious cytotoxicity in the fermentation process, and the normal metabolic consumption of the thalli can be maintained.

Example 6

Acetyl phosphate and AcCoA are prepared by different ways, the reaction is carried out for 2h, the contents of Ara5P and AcCoA are detected after the reaction is finished, the content of AcCoA can also indirectly indicate the synthesis amount of acetyl phosphate, and reactants, enzymes and the addition amount of the reactants are shown in Table 3.

And detecting Ara5P, namely taking 50 mu L reaction liquid, freeze-drying the reaction liquid, respectively deriving for 1h by utilizing 30 mu L methoxylamine hydrochloride and 90 mu L trimethylsilyl trifluoroacetamide, wherein the deriving temperature is 37 ℃, and detecting the content of Ara5P by utilizing GC-MS.

AcCoA was assayed by collecting 50. mu. L reaction solution, terminating the reaction with 10% sulfuric acid solution, removing impurities with a 0.22 μm filter, and performing liquid phase assay with mobile phase A of 0.2M, pH ═ 5 sodium dihydrogenphosphate solution, mobile phase B of acetonitrile, and the maximum absorption peak of AcCoA appearing at 254 nm.

The results of the product content measurements are shown in Table 3.

The results in Table 3 show that, when no aldolase or transaldolase was added to the reaction system, Ara5P was not produced, the amount of AcCoA produced was very small, and the reaction conversion rate was very low; after aldolase or transaldolase and G3P are added into the reaction system, Ara5P can be generated, and the reaction conversion rate is high; after RpiA and Rpe are added simultaneously, the generated Ara5P is converted into acetyl phosphate under the actions of RpiA, Rpe and F/Xpk, and is further converted into AcCoA by phosphate acetyltransferase, and the detection results of Ara5P and AcCoA show that the reaction conversion rate is very high; DHAP and TpiA are added into the system at the same time to replace G3P, and the effect similar to that of direct addition of G3P can be obtained, so that higher conversion rate is achieved.

TABLE 3 Table of substance addition and metabolite production of the system

(concentration unit of reactant or product: mM; amount unit of enzyme: μ g)

Remarking: the buffer solution contains Tris, NaCl and MgCl₂Three major components, pH7.5, 37 ℃.

In addition, in the same reaction system containing reactants such as GA, Phos, G3P, ThDP and the like, respectively: a: no aldolase or transaldolase was added; b: addition of Tal B^F178Y(ii) a C: addition of Tal B^F178YAnd F/Xpk; d: addition of Tal B^F178YF/Xpk and Rpi A; e: addition of Tal B^F178YF/Xpk and Rpi A and Rpe. The content of Ara5P was determined by GC-MS, and the results are shown in the A-E chart of FIG. 2 (the peak time of Ara5P was 37 min). As can be seen from the graphs in FIG. 2 and the corresponding reaction conditions, Ara5P was produced only after addition of aldolase; only when Rpi A and Rpe are added simultaneously, the produced Ara5P is decomposed more completely by phosphoketolase.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and the like that are within the spirit and principle of the present invention are included in the present invention.

Claims

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.