94 results about "Polyphosphate kinase" patented technology

The invention relates to a recombinant pichia pastoris strain capable of producing astaxanthin at high yield as well as a construction method and application of the recombinant pichia pastoris strain. The recombinant pichia pastoris strain is obtained by expressing ATP (adenosine triphosphate) citrate lyase (ACL) and acetyl-CoA synthase (ACS) or phosphoketolase (PK) and phosphotransacetylase (PTA), choline kinase (CK), inositol polyphosphate kinase (IPK) and vitreoscilla hemoglobin (VHB) in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified strain PP-LC2. The astaxanthin production performance of the recombinant strain is verified on the basis of comparison of precursor supply pathways, IUP pathway construction and improvement of the oxygen supply capacity of the engineering strain, and the astaxanthin production capacity of the pichia pastoris is further improved. The construction method of the recombinant pichia pastoris is simple, the synthesis of astaxanthin can be better promoted, the engineering strain can efficiently synthesize the astaxanthin by utilizing methanol through amplification fermentation of a 5L fermentation tank, and industrial production is facilitated.

The invention discloses a polyphosphate kinase mutant, a biological material, a catalyst and application, and belongs to the field of protein engineering, and compared with a wild enzyme, the mutant has one or more amino acid residue mutations in the 307th site, the 311th site, the 320th site, the 408th site, the 419th site and the 431th site. The mutant disclosed by the invention is obtained by combining a semi-rational design strategy with active site region prediction and mutation library design screening, and the enzyme activity and the relative yield are remarkably improved: the efficiency of catalytic synthesis of GTP by using the preferable mutant Y431P / K311F / W408A with GMP as a substrate is 2.6 times that of a wild type enzyme, and the relative yield is improved by 150%; the efficiency of the optimized mutant Y431P / K311F / R419L for catalytically synthesizing GTP by taking GDP as a substrate is 1.9 times that of a wild enzyme, and the relative yield is increased by 82%. The key mutant obtained by screening can realize efficient synthesis from cheap NDP or NMP to high-value NTP, and has an excellent industrial application prospect.

The invention belongs to the technical field of bioengineering, and particularly relates to a polyphosphate kinase mutant and application thereof in synthesis of beta-nicotinamide mononucleotide, and the polyphosphate kinase mutant is obtained by single-point or multi-point mutation of 99th, 186th, 190th and 204th amino acids of polyphosphatase with an amino acid sequence shown as SEQ ID NO.1. The invention also relates to a preparation method of the polyphosphate kinase mutant, and the application of the polyphosphate kinase mutant in synthesis of beta-nicotinamide mononucleotide in synthesis of beta-nicotinamide mononucleotide in synthesis of beta-nicotinamide mononucleotide in synthesis of beta-nicotinamide mononucleotide. Polyphosphokinase derived from Lampropedia Hyalina DSM 16112 is subjected to molecular modification to obtain multiple mutants, the enzyme activity of the mutants is improved by 1.5-14 times compared with that of wild type polyphosphokinase, the preference of the mutants to AMP is improved by 1.08-4.88 times compared with that of the wild type polyphosphokinase, and an ATP regeneration system constructed by the mutants can remarkably reduce the use amount of enzyme and reduce the fermentation capacity and cost; the reaction time can be greatly shortened, the influence of AMP on the enzyme activity is reduced, the requirements of large-scale industrial production of NMN prepared by adopting a biological enzyme method can be met, and the method has a wide application prospect.

The invention discloses a synthetic method of 2 '-fluoro-2'-deoxynucleosid-5 '-triphosphoric acid and application of the 2'-fluoro-2 '-deoxynucleosid-5'-triphosphoric acid. The synthesis method comprises the following steps: reacting in a system containing adenosine triphosphate, 2 '-fluoro-2'-deoxynucleoside, metal ions and deoxynucleoside kinase to obtain 2 '-fluoro-2'-deoxynucleoside-5 '-phosphoric acid; and reacting in a system containing polyphosphoric acid, 2 '-fluoro-2'-deoxynucleoside-5 '-phosphoric acid, metal ions and polyphosphate kinase to obtain 2'-fluoro-2 '-deoxynucleoside-5'-triphosphoric acid. The obtained 2 '-fluoro-2'-deoxynucleoside-5 '-triphosphoric acid can be used for preparing heterologous nucleic acid. The invention provides an economic and green new way for synthesis of 2 '-fluoro-2'-deoxynucleoside-5 '-triphosphate, solves the problems of complex steps, non-environment-friendly reagents and high cost in the existing chemical synthesis method, and provides a thought for preparation of nucleotide derivatives.

The invention belongs to the technical field of soil organic matter improvers, and discloses a soil organic matter improver based on a compound microbial agent and a preparation method thereof, and the soil organic matter improver comprises the following components: a compound enzyme, a modified chitosan microsphere complex, a modified polyelectrolyte, an inhibitor L-methionine sulfoximine, a substrate, an auxiliary factor and the compound microbial agent. A compound enzyme system is used as a core, soil ammonium ions are circularly and efficiently assimilated through glutamine synthetase / glutamate synthetase, and the soil ammonium ions are converted into organic nitrogen; and energy guarantee is provided by virtue of polyphosphate kinase. The modified chitosan microsphere complex is used for protecting and slowly releasing enzyme and a substrate, and controlled release of the inhibitor L-methionine sulfoximine is realized through the modified polyelectrolyte, so that carbon-nitrogen balance of soil is regulated and controlled. The system has a synergistic effect with a compound microbial agent, the problem of soil organic matter reduction caused by excessive nitrogen application is effectively solved, meanwhile, the crop root biomass is remarkably increased, and the soil organic matter content is increased.

The invention discloses a method for biological high-density synthesis of retinaldehyde and a synthetic strain thereof. The synthetic strain is obtained by expressing ATP citrate lyase ACL, acetyl-CoA synthase ACS, choline kinase CK, inositol polyphosphate kinase IPK and vitreoscilla hemoglobin VHb in host bacteria. Wherein the host bacterium is a pichia pastoris gene modified bacterial strain PP-B2C. The retinaldehyde production performance of the recombinant strain is verified on the basis of a precursor supply pathway, an IUP pathway and a dissolved oxygen level, and the retinaldehyde production capacity of the pichia pastoris is further improved. The construction method of the recombinant pichia pastoris is simple, the synthesis of retinaldehyde can be better promoted, and the retinaldehyde can be efficiently synthesized by the engineering strain by utilizing glucose and methanol through amplification fermentation of a 5L fermentation tank, so that the recombinant pichia pastoris is beneficial to industrial production.

2'-deoxycytidine-5'-monophosphate and an enzymatic synthesis method therefor, relating to the technical field of biosynthesis. The synthesis method comprises: constructing a first engineered strain and a second engineered strain; respectively carrying out fermentation culture and protein separation and purification on the first engineered strain and the second engineered strain to obtain high-purity deoxycytidine kinase and high-purity polyphosphate kinase; using 2'-deoxycytidine as a deoxycytidine substrate and adenosine triphosphate as a phosphate donor, cyclically feeding polyphosphate, and keeping the ratio of adenosine triphosphate to polyphosphate to a preset ratio to form an enzymatic reaction system; and performing an enzymatic reaction to obtain 2'-deoxycytidine-5'-monophosphate. High-concentration deoxycytidine monophosphate can be quickly and effectively produced, so that pollution hazards caused by a chemical synthesis method can be solved, and the input amount of adenosine triphosphate as a phosphate donor in a biosynthesis system can also be reduced, thereby reducing production costs.

The invention relates to an engineering bacterium for synthesizing mannitol by using fructose and nicotinamide ribose and application thereof. The invention provides genetic recombination escherichia coli for synthesizing D-mannitol by using D-fructose and nicotinamide ribose, which comprises the following modifications: insertion or enhanced expression of a gene cluster 1, the gene cluster 1 comprises a D-mannitol dehydrogenase coding gene MDH and a formate dehydrogenase coding gene FDH; deletion or inhibitory expression of a gene cluster 2, wherein the gene cluster 2 comprises a 6-phosphofructokinase 1 coding gene pfkA; and insertion or enhanced expression of a gene cluster 3, wherein the gene cluster 3 comprises a nicotinamide ribokinase coding gene NRK, a nicotinamide mononucleotide adenosine transferase coding gene NMNAT, a polyphosphate kinase coding gene PPK2 and a pyrophosphohydrolase coding gene IPPa. The strain disclosed by the invention can effectively synthesize high-purity D-mannitol by using D-fructose as a production raw material and nicotinamide ribose as a co-substrate through an enzyme catalytic reaction under the condition of not additionally adding NAD < + > or NADP < + >, so that the production cost is greatly reduced.

The invention discloses an application of engineered citrobacter freundii in fermentation synthesis of a stable body. The engineered citrobacter freundii is citrobacter freundii overexpressing a polyphosphate kinase gene Ppk1. The stable body prepared by fermenting the engineered citrobacter freundii is rich in polyphosphates and polyamines, particularly, the content of the polyamines can be up to 21wt%, the polyphosphates in the stable body can store the polyamines in an ionic bond combination mode, the combined polyamines are insoluble in water, a natural anion coating is generated, and the stability of the stable body is improved. And the sustained-release tablet is not easily taken by intestinal epithelial cells, so that the sustained-release tablet has a certain sustained-release effect. The medicine prepared by taking the compound as a main active ingredient shows a remarkable curative effect in ulcerative colitis treatment, has the comprehensive effects of reducing oxidative stress, enhancing the anti-inflammatory ability of a body and regulating immunity, is suitable for patients of all ages, and has outstanding application value and market prospect.

The invention belongs to the technical field of biology, and particularly relates to a method for synthesizing D-tagatose through multi-enzyme cascade catalysis, lactose is used as a substrate, and the D-tagatose is synthesized through synchronous cascade catalysis of beta-galactosidase, L-arabinose isomerase, glucose isomerase, fructokinase, tagatose-1, 6-diphosphate aldolase, polyphosphate kinase and phosphatase. According to the invention, the intermediate product D-glucose is continuously converted into D-tagatose, so that the conversion rate of D-tagatose is increased, a new method is provided for synthesizing D-tagatose from lactose, and certain theoretical basis and technical support are provided for realizing industrial production of D-tagatose with high added value.

The invention discloses a method for synthesizing D-psicose through multi-enzyme co-immobilization cascade combination, and belongs to the field of enzyme engineering. According to the method provided by the invention, D-psicose-3-epimerase, L-rhamnokinase, polyphosphate kinase and prepared nitrilotriacetic acid modified magnetic nanoparticles are mixed and incubated to obtain the co-immobilized enzyme. According to the efficient co-immobilized enzyme provided by the invention, the stability is improved, the efficient co-immobilized enzyme is easily separated from an enzymolysis system, the repeated utilization of the immobilized enzyme can be realized, and the production cost is reduced. When the co-immobilized enzyme is applied to preparation of D-psicose, the raw material cost of an enzyme preparation is greatly reduced, and the co-immobilized enzyme has relatively high industrial application value.

The present invention discloses a polyphosphate kinase mutant and its application in glutamine synthesis, belonging to the field of biotechnology. The present invention has screened a polyphosphate kinase mutant with significantly improved enzyme activity. The catalytic activity of this mutant enzyme has increased by 182.2% compared to the original enzyme, enabling efficient regeneration of ATP and reducing the cost of biocatalysis. Using the polyphosphate kinase mutant constructed in the present invention for the production of glutamine significantly improves the sustainability of ATP regeneration. The glutamine yield can reach 94.4 mM after 8 h of reaction, and the conversion rate is 94.4%. The yield of glutamine produced by coupling with the original enzyme is increased by 41.5%.

The invention provides a yeast rich in polyphosphate and application of the yeast in synthesis of adenosine triphosphate, and the content of polyphosphate is 15-35 wt% based on the dry matter mass of the yeast rich in polyphosphate; according to the method disclosed by the invention, the yeast is subjected to phosphorus starvation culture and then is subjected to phosphorus supplementary culture, so that the yeast is enriched with polyphosphates, and the yeast rich in polyphosphates is obtained; then, the yeast rich in polyphosphate is used as a biocatalyst to synthesize ATP, that is, self-related enzyme systems, mainly including adenosine kinase, endo-polyphosphate kinase and excision-polyphosphate kinase, in the yeast rich in polyphosphate are used for replacing exogenous polyphosphate kinase and adenosine kinase, and in addition, ATP is synthesized by using the yeast rich in polyphosphate as a biocatalyst. According to the method for synthesizing ATP by using the yeast rich in polyphosphate as the biocatalyst, exogenous adenosine kinase and exogenous polyphosphate kinase do not need to be additionally added, the cost is low, the efficiency of converting adenosine into ATP is high, the batch-to-batch stability of the yeast is realized, and the enzyme activity of the self-related enzyme system is high.

The invention discloses a method for catalytically synthesizing adenosine triphosphate by using microcrystalline cellulose carrier purification immobilized enzyme, which comprises the following steps: preparing bacterial suspension by using escherichia coli wet thalli and Tris-HCL buffer solution, crushing, centrifuging, and taking supernate to obtain crude enzyme; adding a microcrystalline cellulose carrier and the wet thalli into the crude enzyme liquid, and washing the immobilized enzyme after immobilization to obtain the immobilized enzyme; the method comprises the following steps: adding immobilized enzyme adenosine kinase and immobilized polyphosphate kinase into a substrate system with the volume of 1L according to a weight ratio, and starting a catalytic reaction under the conditions that the pH is 5.5-7.5 and the temperature is 20-40 DEG C; after catalysis is finished, the immobilized enzyme adenosine kinase and the immobilized polyphosphate kinase are subjected to solid-liquid separation with reaction liquid through a Buchner funnel, the immobilized enzyme continues to be subjected to catalytic reaction, and the method for preparing the immobilized enzyme is simple in mode, low in cost, easy to industrially produce, high in stability and capable of being repeatedly used for many times, and products are easy to separate.

The present invention relates to a method for producing cytidine 5′-monophospho-N-acetyl-neuraminic acid (CMP-Neu5Ac, 1) from low-cost substrates N-acetyl-D-glucosamine (GlcNAc), pyruvate, cytidine and polyphosphate in a single reaction mixture with a set of optionally immobilized or optionally co-immobilized enzymes comprising N-acylglucosamine 2-epimerase (AGE), an N-acetylneuraminate lyase (NAL), an N-acylneuraminate cytidylyltransferase (CSS), a uridine kinase (UDK), a uridine monophosphate kinase and a polyphosphate kinase 3 (PPK3). Further, said process may be adapted to produce Neu5Acylated i.e. sialylated biomolecules and biomolecules including a saccharide, a peptide, a protein, a glycopeptide, a glycoprotein, a glycolipid, a glycan, an antibody, and a glycoconjugate, in particular, an antibody drug conjugate, and a carbohydrate conjugate vaccine, or a flavonoid.

The invention relates to the field of polyphosphate kinase, and discloses polyphosphate kinase 2 as well as expression and application thereof. The invention provides a coding gene of polyphosphate kinase 2. The nucleotide sequence of the coding gene is shown as SEQ ID NO. 1. According to the polyphosphokinase 2, efficient soluble expression of the polyphosphokinase 2 is realized by modifying and optimizing a gene sequence, so that the polyphosphokinase 2 obtained by induced expression has relatively high catalytic activity on AMP / ADP and wide adaptability of taking cheap polyphosphoric acid as a phosphoric acid donor, and can stably realize efficient conversion from AMP / ADP to ATP; the method can be used for establishing a low-cost and high-efficiency ATP regeneration system.

The invention relates to a biosynthesis method of pyridoxal phosphate or pyridoxine phosphate, and belongs to the technical field of biosynthesis. In order to solve the existing problem of serious environmental pollution, the invention provides a biosynthesis method of pyridoxal phosphate or pyridoxine phosphate, which comprises the following steps: in the presence of polyphosphate, magnesium salt, ATP (adenosine triphosphate), pyridoxal kinase and polyphosphate kinase, performing phosphorylation reaction on pyridoxal or pyridoxine and acid salts thereof, thereby obtaining the pyridoxal phosphate or pyridoxine phosphate. The temperature of the phosphorylation reaction is controlled to be 30-38 DEG C, after the reaction is finished, protein separation is conducted on reaction liquid, mother liquor is separated, the compound pyridoxal phosphate or pyridoxine phosphate shown in the formula I is obtained, and the mother liquor is recycled and reused. According to the method, a large amount of ammonium acetophosphate does not need to be additionally added, operation is simpler, mother liquor can be recycled, a large amount of waste water is prevented from being discharged, environment friendliness is better achieved, and the yield is high.

The application discloses a polyphosphate kinase mutant, a preparation method and application thereof, and wild-type polyphosphate kinase from Pseudomonas aeruginosa Pseudomonas aeruginosa PAO1, The active site is mutated by using a directed evolution method, and a polyphosphate kinase mutant with high activity in catalyzing synthesis of ATP is obtained, the relative activity of the mutant to a substrate ADP or AMP is much higher than that of a wild type, and the mutation is at least one of D307M, W408A and Y431P, or at least one of E304V, P331S and Y431P. The application realizes efficient synthesis from a cheap substrate to a high-value product, is helpful to industrial production of ATP, and has a good application prospect in the fields of medicine and the like.

The present invention relates to a method for producing α-sialyl-β-D-galactoside saccharides, particularly α-sialyl-(2→3)-β-D-galactoside saccharides and α-sialyl-(2→6)-β-D-galactoside saccharides, from a β-D-galactoside saccharide, a sialic acid donor, and an enzyme with β-galactoside α-sialyltransferase activity. The enzymes with β-galactoside α-sialyltransferase activity used herein do not exhibit catalytic activity towards the hydrolysis of cytidine 5'-monophospho-N-acetyl-neuraminic acid to cytidine and N-acetyl-neuraminic acid. The method can be performed in vitro and in vivo using a genetically engineered cell comprising a nucleic acid encoding said enzyme. Further, said process may be adapted to produce the sialic acid donor CMP-Neu5Ac from low-cost substrates N-acetyl-D-glucosamine (GlcNAc), pyruvate, a cytidine phosphate (CMP, CDP or CTP) and polyphosphate in a single reaction mixture with a set of optionally immobilized or optionally co-immobilized enzymes comprising N-acylglucoamine 2-epimerase (AGE), an N-acetylneuraminate lyase (NAL), an N-acylneuraminate cytidylyltransferase (CSS), optional a uridine kinase (UDK), a uridine monophosphate kinase and a polyphosphate kinase 3 (PPK3).

The invention discloses a high-yield heparosan gene engineering bacterium and application thereof, and belongs to the technical field of gene engineering and metabolic engineering. The invention discloses a high-yield heparosan genetically engineered bacterium which is prepared by the following steps: taking corynebacterium glutamicum BDH as an original strain, and knocking out a glucose-6-phosphate dehydrogenase gene and a lactic dehydrogenase gene; a fructose-1, 6-diphosphate aldolase gene is weakened; according to the invention, a transglutamine-6-phosphate fructose transaminase gene, a nucleoside diphosphate kinase gene, an NADH oxidase gene, a glutamine synthetase gene and a polyphosphate kinase gene are over-expressed. The gene engineering bacterium constructed by the invention can be used for producing 23.05 g / L of heparosan.

The invention relates to the technical field of enzymes, in particular to a BsBacD enzyme mutant, an enzyme composition and a preparation method of tyrosine dipeptide. The method comprises the following steps: rationally designing BsBacD enzyme, replacing ADP (adenosine diphosphate or salt thereof) with relatively cheap ATP (adenosine triphosphate or salt thereof) or relatively cheap AMP (adenosine monophosphate or salt thereof), and introducing polyphosphate kinase ChPPK2 capable of regenerating ATP by taking AMP as a substrate to synthesize tyrosine dipeptide; parameters such as reaction pH, magnesium chloride concentration, substrate tyrosine concentration and the like are optimized, and a low-cost and efficient enzymatic synthesis process of three tyrosine dipeptides such as propyl tyrosine dipeptide, glycyl tyrosine dipeptide and sericin tyrosine dipeptide is developed.

The present application relates to the technical field of bioengineering, and particularly relates to a polyphosphate kinase mutant with high catalytic activity and application thereof in synthesis of glutathione. The present application provides a polyphosphate kinase mutant with high catalytic activity, which is characterized in that the polyphosphate kinase mutant is mutated from an amino acid sequence shown in SEQ ID NO: 2 into one of the following: (1) single mutation is made to the 234th amino acid in the amino acid sequence shown in SEQ ID NO: 2; (2) double mutations are made to the 234th and 166th amino acids in the amino acid sequence shown in SEQ ID NO: 2; wherein a nucleotide sequence of a coding gene coding the amino acid sequence shown in SEQ ID NO: 2 is as shown in SEQ ID NO: 1. ch PPK-N234A and double-site mutation polyphosphate kinase mutant ch PPK-N234A / K166D has high activity, and the enzyme activity is increased by 4.19 times and 6.12 times, respectively, compared with wild-type polyphosphate kinase.

The invention discloses a method for improving the yield of L-threonine by enhancing Escherichia coli electronic respiration transfer and energy transfer efficiency, and belongs to the technical field of genetic engineering. According to the invention, a cytochrome bo oxidase subunit CyoA mutant derived from escherichia coli, a cytochrome bo oxidase subunit CyoB mutant derived from pseudomonas aeruginosa, a cytochrome bd oxidase cydAB gene cluster derived from escherichia coli and a polyphosphate kinase PPK mutant derived from radiation-resistant coccus are introduced, amn genes are knocked out, and a cytochrome bo oxidase gene is obtained. The electron transfer efficiency in escherichia coli and the production efficiency of energy ATP (adenosine triphosphate) are enhanced, and the production efficiency of L-threonine and the carbon yield are further improved. Finally, the obtained engineering strain ECTHRS-5 is subjected to fed-batch fermentation in a 5L fermentation tank to accumulate 160.8 g / L of L-threonine, and the sugar-acid conversion rate is 0.62 g / g.

This invention discloses the application of engineered *Citrobacter freundii* in the fermentation synthesis of spermine. The engineered *Citrobacter freundii* is a strain overexpressing the polyphosphate kinase gene Ppk1. The engineered *Citrobacter freundii* is inoculated into a fermentation medium for fermentation culture to synthesize spermine. The synthesis and extraction method of this invention for producing spermine is simple to operate, highly reproducible, low in cost, and yields a high spermine content in the fermented bacterial cells, accounting for 12.42% of the bacterial dry weight. The extracted spermine has a purity as high as 90.26%.

The application belongs to the technical field of biology and specifically relates to a method for synthesizing D-tagatose through multi-enzyme cascade catalysis, which uses lactose as a substrate and utilizes beta-galactoside enzyme, L-arabinose isomerase, glucose isomerase, fructokinase, tagatose-1,6-diphosphate aldolase, polyphosphate kinase and phosphatase for synchronous cascade catalysis to synthesize D-tagatose. The method improves the conversion rate of D-tagatose by continuously converting the intermediate product D-glucose into D-tagatose, provides a new method for synthesizing D-tagatose from lactose, and provides certain theoretical basis and technical support for realizing the industrialized production of high-value-added D-tagatose.

The present disclosure discloses a method for efficient catalytic synthesis of PAPS based on constructing an ATP regeneration system, and belongs to the technical field of bioengineering. Efficient production of PAPS is realized through microbial recombination expression and artificial construction of PAPS bifunctional synthetase. On the basis, an ATP regeneration system coupling with polyphosphate kinase from Corynebacterium glutamicum and Mycobacterium tuberculosis can be used for recovering two byproducts: pyrophosphoric acid and ADP at the same time, the equivalent conversion of a substrate and a product is realized, the PAPS generated in a catalysis system has high purity, and the sulfonic acid group donation in most sulfonic acid transfer reactions can be realized.

The present disclosure relates to biocatalytic ligation methods for producing oligonucleotides; and to fusion polypeptides for use in said methods. In particular, the disclosure relates to biocatalytic ligation methods incorporating ATP regeneration, and to fusion polypeptides comprising a polyphosphate kinase domain and an ATP-dependent nucleic acid ligase domain.

This invention discloses an enzymatic method for preparing (S)-adenosylmethionine. The method uses adenosine diphosphate (ADP) and L-methionine as substrates, reacting them under the combined action of polyphosphate kinase (Ppk) and adenosylmethionine synthase (MAT) to obtain (S)-adenosylmethionine. This method produces a product with high chiral purity and high atom economy, making it more suitable for industrial production.

The present disclosure relates to biocatalytic ligation methods for generating oligonucleotides and fusion polypeptides for use in the methods. In particular, the disclosure relates to biocatalytic ligation methods that incorporate ATP regeneration and fusion polypeptides that include a polyphosphate kinase domain and an ATP-dependent nucleic acid ligase domain.

The invention discloses a vidarabine monophosphate biosynthesis method based on an ATP (adenosine triphosphate) regeneration system, and belongs to the technical field of biomedical engineering. According to the method, riboside arabinoside and ATP serve as reaction substrates, the target product riboside arabinoside monophosphate is synthesized through double-enzyme catalysis, double enzymes are deoxynucleoside kinase and polyphosphate kinase, the deoxynucleoside kinase is mainly used for catalyzing riboside arabinoside to synthesize riboside arabinoside monophosphate, and the polyphosphate kinase is used for catalyzing riboside arabinoside to synthesize riboside arabinoside monophosphate. Polyphosphokinase is mainly used for catalyzing ADP (adenosine diphosphate) and polyphosphoric acid or polyphosphate to regenerate to obtain ATP (adenosine triphosphate). The method can be used for a pure enzyme reaction system, a crude enzyme liquid reaction system or a resting cell catalysis system. The method disclosed by the invention is environment-friendly, realizes low-cost efficient conversion of a substrate through coupled ATP regeneration, plays a significant role in promoting biosynthesis of arabinoside monophosphate series nucleoside drugs, and has application potential and economic value.