CN117070582A - Method for synthesizing beta-nicotinamide mononucleotide by hypoxanthine-guanine phosphoribosyl transferase - Google Patents
Method for synthesizing beta-nicotinamide mononucleotide by hypoxanthine-guanine phosphoribosyl transferase Download PDFInfo
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- 230000002194 synthesizing effect Effects 0.000 title claims abstract description 13
- PQGCEDQWHSBAJP-TXICZTDVSA-N 5-O-phosphono-alpha-D-ribofuranosyl diphosphate Chemical compound O[C@H]1[C@@H](O)[C@@H](O[P@](O)(=O)OP(O)(O)=O)O[C@@H]1COP(O)(O)=O PQGCEDQWHSBAJP-TXICZTDVSA-N 0.000 claims abstract description 44
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 claims abstract description 36
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- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 1
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- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
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- 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
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/30—Nucleotides
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Abstract
The invention discloses a method for synthesizing beta-nicotinamide mononucleotide by using hypoxanthine-guanine phosphoribosyl transferase, which takes nicotinamide and 5-phosphoribosyl-1-pyrophosphate as substrates, and the nicotinamide mononucleotide is generated by condensing the nicotinamide and the 5-phosphoribosyl-1-pyrophosphate by using a hypoxanthine-guanine phosphoribosyl transferase catalytic system, wherein the catalytic system consists of hypoxanthine-guanine phosphoribosyl transferase and a PRPP (PRPP) synthesis system. The method for synthesizing the beta-nicotinamide mononucleotide by using the hypoxanthine-guanine phosphoribosyl transferase as the key enzyme has the advantages of small molecular weight of the key enzyme, good thermal stability, strong enzyme engineering operability, wide sources, simple reaction steps, mild reaction conditions, high catalytic rate, environment friendliness, simplicity and convenience in enzyme preparation and the like, and has good application and development prospects in the field of industrial synthesis of the beta-nicotinamide mononucleotide and related products thereof.
Description
Technical Field
The invention belongs to the technical field of biological enzyme engineering, and in particular relates to a method for synthesizing beta-nicotinamide mononucleotide by using hypoxanthine-guanine phosphoribosyl transferase, which takes hypoxanthine-guanine phosphoribosyl transferase as a catalyst to catalyze condensation of nicotinamide and 5-phosphoribosyl-1-pyrophosphoric acid to obtain beta-nicotinamide mononucleotide in a stereoselective manner.
Background
Beta-nicotinamide mononucleotide (beta-nicotinamide mononucleotide, NMN) is a precursor of nicotinamide adenine dinucleotide (Nicotinamide adenine dinucleotide, NAD, also known as coenzyme I), an important intermediate metabolite in cells. NAD is difficult to enter cells directly, NMN is a direct precursor to NAD synthesis, is well tolerated, and is capable of preventing age-related physiological decline (Mills KF, yoshida S, stein LR, et al Long-Term Administration of Nicotinamide Mononucleotide Mitigates Age-Associated Physiological Decline in Mice. Cell Metab.2016,24, 795-806), and is therefore the preferred substance for NAD supplementation. In addition, NMN has been shown to be effective in treating high fat diet-induced type 2 diabetes by reversing mitochondrial dysfunction associated with aging (Haigis MC, mostonplavsky R, haigis KM, et al SIRT4 inhibits glutamate dehydrogenase and opposes the effects of calorie restriction in pancreatic beta cells 2006,126, 941-954). However, natural NMN is extremely low in the content of organisms and cannot be obtained by separation and extraction. At present, the traditional NMN preparation method is prepared by a chemical synthesis way, but the cost for preparing high-purity NMN is high due to the fact that the stereoselectivity is difficult to control, the yield is low, and the product taking the NMN as a main active ingredient is quite expensive. Compared with chemical synthesis, the biological enzyme method for synthesizing NMN has the advantages of high efficiency and environmental protection, no organic solvent residue and no chiral problem. At present, nicotinamide phosphoribosyl transferase for catalyzing NMN synthesis is limited by factors such as large molecular weight, complex structure, difficult directed evolution and the like, and a novel efficient phosphoribosyl transferase for synthesizing NMN is still to be developed.
Hypoxanthine-guanine phosphoribosyl transferase (HGPRT), belonging to the family of 6-oxopurine phosphoribosyl transferases (PRTase), has the biological function of catalyzing the condensation of guanine/Hypoxanthine with 5-phosphoribosyl-1-pyrophosphate (PRPP) to produce guanine/inosine monophosphate (GMP/IMP), which is a key enzyme in the salvage synthesis pathway of guanine and Hypoxanthine.
Disclosure of Invention
The invention aims to provide a method for synthesizing beta-Nicotinamide Mononucleotide (NMN) by using hypoxanthine-guanine phosphoribosyl transferase, which is a method for obtaining NMN by condensing Nicotinamide (NAM) and 5-phosphoribosyl-1-pyrophosphate (PRPP) by using hypoxanthine-guanine phosphoribosyl transferase stereoselectivity, and is an expansion application of hypoxanthine-guanine phosphoribosyl transferase substrate spectrum.
The method provided by the invention takes NAM and PRPP as raw materials and Mg as raw materials 2+ NMN is obtained as cofactor by enzymatic catalysis of hypoxanthine-guanine phosphoribosyl transferase.
The method comprises the following specific steps: takes NAM and PRPP as raw materials and Mg 2+ As cofactor, under the catalysis of hypoxanthine-guanine phosphoribosyl transferase in an enzyme catalysis system, NMN is obtained by condensation, and the reaction formula is as follows:
in the reaction process, NAM and PRPP are used as substrates, but the PRPP is expensive and easy to decompose, and the PRPP is synthesized by the PRPP synthesis system, so that the reaction cost can be obviously reduced.
In the invention, the PRPP synthesis system is as follows: ribose kinase and phosphoribosyl pyrophosphokinase as key enzymes of the system, ATP and D-ribose as substrates, mg 2+ PRPP synthesis system as cofactor.
Therefore, the NMN enzyme catalytic synthesis system also comprises a PRPP synthesis system, and the enzyme system comprises hypoxanthine-guanine phosphoribosyl transferase, ribose kinase, phosphoribosyl pyrophosphatase and magnesium chloride. The hypoxanthine-guanine phosphoribosyl transferase, ribose kinase and phosphoribosyl pyrophosphatase in the enzyme catalytic system are all free pure enzymes after purification. Adding ATP, D-ribose, NAM and PRPP into the enzyme reaction system, and controlling the pH and temperature under the condition of Mg 2+ As cofactors, ATP phosphorylates D-ribose to 5-ribose phosphate under the catalysis of ribokinase, and further phosphorylates 5-ribose pyrophosphate kinase under the catalysis of ribokinasePhosphoribosyl phosphate is converted into PRPP, and finally, hypoxanthine-guanine phosphoribosyl transferase condenses NAM with PRPP to generate NMN, and the reaction formula is as follows:
specifically, the nucleotide sequence for encoding the hypoxanthine-guanine phosphoribosyl transferase is derived from GenBank, the number is U88876.1, and the hypoxanthine-guanine phosphoribosyl transferase is obtained by total gene synthesis of mycobacterium tuberculosis (Mycobacterium tuberculosis) through codon optimization, as shown by MtHGPRT-DNA (SEQ.No.1) in a sequence table.
Specifically, the amino acid sequence of the hypoxanthine-guanine phosphoribosyl transferase is shown as MtHGPRT-AA (SEQ No. 2) in a sequence table.
As known to those skilled in the art, the nucleotide sequence of the hypoxanthine-guanine phosphoribosyl transferase gene of the present invention can also be any other nucleotide sequence encoding the amino acid sequence shown as MtHGPRT-AA (SEQ. No.2) in the sequence Listing.
Any nucleotide sequence obtained by substitution, certainty or insertion treatment of one or more nucleotides with respect to the nucleotide sequence shown in MtHGPRT-DNA is within the scope of the present invention as long as it has homology of 90% or more with the nucleotides.
Any amino acid sequence shown in MtHGPRT-AA is deleted, inserted or substituted with one or more amino acids and has NMN synthesis activity, and still falls into the protection scope of the invention.
For hypoxanthine-guanine phosphoribosyl transferase, any other source of isoenzyme having NMN synthesizing activity is within the scope of the invention.
Specifically, the nucleotide sequence encoding the ribokinase is derived from GenBank, accession No.: 948260, E.coli (strain K12) ribokinase), is obtained by total gene synthesis from biotechnology, as shown in RBKS-DNA (SEQ. No. 3) of the sequence Listing.
Specifically, the amino acid sequence of the ribokinase is shown as RBKS-AA (SEQ. No. 4) in the sequence table.
Specifically, the nucleotide sequence encoding the phosphoribosyl pyrophosphate kinase is derived from GenBank, accession number: 885993, as shown in MtPRS-DNA (SEQ. No. 5) in the sequence Listing, is Mycobacterium tuberculosis (Mycobacterium tuberculosis) phosphoribosyl pyrophosphate kinase, obtained by total gene synthesis of biotechnology company.
Specifically, the amino acid sequence of the phosphoribosyl pyrophosphate kinase is shown as MtPRS-AA (SEQ. No.6) in a sequence table.
Preferably, in the catalytic system, the addition amount of the ribose kinase is 0.05-3.0mg/ml; the addition amount of phosphoribosyl pyrophosphokinase is 0.1-6.0mg/ml; the addition amount of hypoxanthine-guanine phosphoribosyl transferase is 0.1-5.0mg/ml.
In the catalytic system, the addition amount of the substrate NAM is 0.5-30mM; the addition amount of PRPP is 0.75-45mM; cofactor Mg 2+ The addition amount of (C) is 0.1-20mM. The addition amount of the substrate ATP in the PRPP synthesis system is 1.0-30mM; the addition amount of D-ribose is 0.5-10mM.
Preferably, in the enzyme catalytic system, the reaction temperature is 25-37 ℃ and the reaction time is 2-20h; more preferably, the temperature is 30-37 ℃ and the time is 4-14h.
Preferably, the pH value of the reaction is controlled to be 6 to 9, the pH is controlled to be lowered by sodium hydroxide, and the pH is controlled to be raised by formic acid.
The beneficial effects of the invention are mainly as follows: provides a method for catalyzing NAM and PRPP condensation to obtain NMN by taking hypoxanthine-guanine phosphoribosyl transferase as key enzyme, which has not been reported at present. Compared with nicotinamide phosphoribosyl transferase which is a key enzyme for biosynthesis of NMN at present, the hypoxanthine guanine phosphoribosyl transferase has the advantages of small molecular weight, easiness in carrying out related operation of enzyme engineering, strong thermal stability, high heterologous expression efficiency of escherichia coli, easiness in purification preparation, high catalytic reaction speed, high catalytic efficiency and the like; meanwhile, the reaction is carried out at the normal temperature with the pH value close to neutrality, the reaction does not need to be participated by transition metal and organic solvent, the method also has the advantages of mild reaction condition, environmental friendliness and the like, has good industrial application development prospect, almost all organisms including prokaryotes and the like have hypoxanthine-guanine phosphoribosyl transferase, has more abundant biological sources than nicotinamide phosphoribosyl transferase, and provides a rich source for screening efficient NMN synthetase.
Drawings
FIG. 1 is a SDS-PAGE of purified hypoxanthine-guanine phosphoribosyl transferase.
Fig. 2 is a NAM blank high performance liquid chromatogram.
FIG. 3 is a high performance liquid chromatogram of the reaction solution after 3.5 hours of reaction in the catalytic system of hypoxanthine-guanine phosphoribosyl transferase.
Detailed Description
The invention is further described below with reference to the drawings and specific examples. It should be understood that the following examples are illustrative of the present invention and are not intended to limit the scope of the present invention.
The experimental methods in the present invention are conventional methods unless otherwise specified.
The plasmid extraction kit is purchased from Tiangen biochemical technology (Beijing) limited company; e.coli DH 5. Alpha. E.coli BL21 (DE 3) and the like are purchased from the division of biological engineering (Shanghai); pre-stained protein Maker was purchased from New Saimei Biotechnology Inc. of Suzhou.
Common reagents used in the present invention include substrates purchased from Ara Ding Huaxue reagent Inc., shanghai Seiyaku Biotechnology Inc., national pharmaceutical Condition chemical Co., ltd.
EXAMPLE 1 expression of hypoxanthine-guanine phosphoribosyl transferase
The sequence of coding mycobacterium tuberculosis (Mycobacterium tuberculosis) hypoxanthine-guanine phosphoribosyl transferase gene (GenBank: U88876.1) is optimized by a codon (the sequence is shown as SEQ ID NO. 1), and is fully synthesized by a biological engineering (Shanghai) limited company and then is connected into a pET-28a (+) vector to construct a MtHGPRT-pET-28a (+) plasmid, and after sequencing and verifying the sequence, the sequence is transformed into E.coli BL21 (DE 3) competent cells by heat shock to obtain hypoxanthine-guanine phosphoribosyl transferase expressionEngineering bacteria, which are prepared by picking single colony from LB plate medium containing 50 mug/ml kanamycin, inoculating to LB liquid medium containing 50 mug/ml kanamycin, shaking and culturing at 37deg.C shaking table 200rpm for 12 hr, transferring to 3L liquid LB medium for expansion culture, shaking and culturing at 37deg.C shaking table 200rpm for 8 hr, and collecting the optical density OD of the culture solution 600 When the temperature reaches 0.6, the temperature is reduced to 16 ℃, IPTG solution with the final concentration of 0.7mM is added for induction expression for 16h, the culture solution is centrifuged at 8000rpm for 10min, the supernatant culture solution is discarded, and the thalli are preserved at-20 ℃ for standby.
EXAMPLE 2 expression of ribokinase
The coding E.coli (Escherichia coli (strain K12)) ribose kinase gene (GenBank: 948260) sequence is subjected to codon optimization (the sequence is shown as SEQ ID NO. 3), is fully synthesized by a biological engineering (Shanghai) limited company and then is connected into a pET-28a (+) vector to construct an RBKS-pET-28a (+) plasmid, after sequencing and verifying the sequence, the RBKS-pET-28a (+) vector is transformed into E.coli BL21 (DE 3) competent cells by heat shock, hypoxanthine-guanine phosphoribosyl transferase expression engineering bacteria are obtained, single colony is selected from LB plate medium containing 50 mug/ml kanamycin, inoculated into LB liquid medium containing 50 mug/ml kanamycin, shake-cultured for 12 hours at 37 ℃, transferred into 1.5L liquid LB medium for expansion, shake-cultured for 8 hours at 200rpm at 37 ℃, and when the optical density OD of the culture solution is obtained 600 When the temperature reaches 0.6, the temperature is reduced to 16 ℃, IPTG solution with the final concentration of 0.2mM is added for induction expression for 16h, the culture solution is centrifuged at 8000rpm for 10min, the supernatant culture solution is discarded, and the thalli are preserved at-20 ℃ for standby.
EXAMPLE 3 expression of phosphoribosyl pyrophosphate kinase
The coding mycobacterium tuberculosis (Mycobacterium tuberculosis) phosphoribosyl pyrophosphate kinase gene (GenBank: 885993) sequence is subjected to codon optimization (the sequence is shown as SEQ ID NO. 5), is fully synthesized by a biological engineering (Shanghai) limited company and is connected with a pET-28a (+) vector to construct a MtPRS-pET-28a (+) plasmid, after sequencing and verifying the sequence, the sequence is transformed into E.coli BL21 (DE 3) competent cells by heat shock to obtain hypoxanthine-guanine phosphoribosyl transferase expression engineering bacteria, and the hypoxanthine-guanine phosphoribosyl transferase expression engineering bacteria are obtained by using LB plate culture medium containing 50 mug/ml kanamycinPicking single colony, inoculating to LB liquid medium containing 50 μg/ml kanamycin, shake culturing at 37deg.C shaking table 200rpm for 12 hr, transferring to 1.5L liquid LB medium, shake culturing at 37deg.C shaking table 200rpm for 8 hr, and collecting culture medium with optical density OD 600 When the temperature reaches 0.6, the temperature is reduced to 16 ℃, IPTG solution with the final concentration of 0.4mM is added for induction expression for 16h, the culture solution is centrifuged at 8000rpm for 10min, the supernatant culture solution is discarded, and the thalli are preserved at-20 ℃ for standby.
EXAMPLE 4 purification of hypoxanthine-guanine phosphoribosyl transferase, ribokinase and phosphoribosyl pyrophosphate kinase
3g of hypoxanthine-guanine phosphoribosyl transferase expression engineering bacteria or ribose kinase, phosphoribosyl pyrophosphatase bacterial cells are resuspended in 20ml lysate (10 mM imidazole, 50mM Tris-HCl,500mM NaCl,10% glycerol, 1% Tween-20 pH 8.0). Shaking, adding lysozyme (1 mg/ml), ice-bathing for 40min, ultrasonic crushing for 3 times, 3 min/time, and centrifuging at 14000rpm for 15min at intervals of 15min to obtain supernatant as crude enzyme solution. Ni-IDA protein purification magnetic beads are used as purification materials, 5ml of 10% magnetic bead suspension is used for a single time, 10ml of lysate is used for balancing the Ni-IDA magnetic beads, crude enzyme solution is added after magnetic separation, mixed incubation is carried out for 1h at 4 ℃, the lysate (50 mM imidazole, 50mM Tris-HCl,500mM NaCl,10% glycerol and 1% Tween-20 pH 8.0) is used for eluting and removing unadsorbed proteins, finally elution buffer (500 mM imidazole, 50mM Tris-HCl,500mM NaCl,10% glycerol and 1% Tween-20 pH 8.0) is used for eluting and collecting target proteins, and 5LKpi buffer (50 mM KH) 2 PO 4 ，50mM K 2 HPO 4 pH 7.4) to dialyze the target protein, remove salts and imidazole, SDS-PAGE analysis results show that the purified yields of hypoxanthine-guanine phosphoribosyl transferase, ribokinase and phosphoribosyl pyrophosphatase can reach 10 mg/liter of culture medium, 20 mg/liter of culture medium and 8 mg/liter of culture medium, respectively, and the purity is more than 95% (figure 1).
EXAMPLE 3 Synthesis of NMN by hypoxanthine-guanine phosphoribosyl transferase
The enzyme purified from hypoxanthine-guanine phosphoribosyl transferase, ribokinase and phosphoribosyl pyrophosphate kinase obtained in example 2Adding into the reaction system according to the addition amount of 0.8mg/ml, respectively adding NAM with the final concentration of 0.8mM, PRPP with the final concentration of 0.8mM, ATP with the final concentration of 1.0mM, D-ribose with the final concentration of 0.8mM and MgCl with the final concentration of 10mM with the pH of 50.4 Kpi as buffer solution 2 After 3.5h of reaction at 37℃with constant temperature shaking (666 rpm), the reaction was terminated by adding an equal volume of methanol, and after 15min centrifugation at 14000rpm, the supernatant was taken and fed into a High Performance Liquid Chromatography (HPLC) to analyze the amounts of substrate and product. The HPLC analysis method comprises the following steps: shimadzu high performance liquid chromatography LC-20AT; chromatographic column extension-C18.6 x 250mm; column temperature is 30 ℃; the flow rate is 0.7ml/min; detection wavelength 245nm; mobile phase: 10% of water (100 mM sodium phosphate salt) and 90% of methanol. And calculating the conversion rate and the yield of the NMN catalyzed by the hypoxanthine-guanine phosphoribosyl transferase by using NAM and NMN standard substance concentration curves. The conversion rate can reach 95.6% and the yield is 90.5% through calculation. The corresponding high performance liquid chromatograms are shown in fig. 2 and 3. The fluorescence analysis method comprises the following steps: mu.l DMSO (containing 20% acetophenone), 10. Mu.l 2M KOH, 25. Mu.l reaction supernatant were mixed, and after 2min in ice bath, 45. Mu.l 88% formic acid was added and incubated at 37℃for 10min. Then, 40. Mu.l of the reacted solution was added to a 384-well blackboard, and fluorescence was detected at an excitation wavelength of 382nm and an emission wavelength of 445 nm. The yield of xanthine-guanine phosphoribosyl transferase catalyzed NMN is calculated from the NMN standard concentration curve. The yield can reach 89.5% by calculation, which is consistent with the result of HPLC detection.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A method for synthesizing beta-nicotinamide mononucleotide by using hypoxanthine-guanine phosphoribosyl transferase is characterized in that nicotinamide and 5-phosphoribosyl-1-pyrophosphate are taken as substrates, and the beta-nicotinamide mononucleotide is obtained by catalysis of an enzyme catalysis system, wherein the enzyme catalysis system consists of hypoxanthine-guanine phosphoribosyl transferase and a PRPP synthesis system.
2. The method according to claim 1, characterized by the following steps: adopts nicotinamide and 5-phosphoribosyl-1-pyrophosphoric acid as raw materials and Mg 2+ Condensing a substrate to obtain beta-nicotinamide mononucleotide under the catalysis of hypoxanthine-guanine phosphoribosyl transferase in an enzyme catalysis system as a cofactor, wherein the reaction formula is as follows:
the PRPP is synthesized by taking the synthesis of PRPP as a precondition and taking ATP and D-ribose as substrates through a PRPP synthesis system through twice catalytic phosphorylation reactions of ribose kinase and phosphoribosyl pyrophosphate kinase.
3. The method according to claim 1 or 2, wherein the nucleotide sequence encoding an inosine-guanine phosphoribosyl transferase is shown in SEQ.No.1 and the amino acid sequence thereof is shown in SEQ.No.2.
4. The method according to claim 1 or 2, wherein the PRPP synthesis system is: ribose kinase and phosphoribosyl pyrophosphokinase as key enzymes of the system, ATP and D-ribose as substrates, mg 2+ As cofactor, with Mg 2+ As cofactor, ATP phosphorylates D-ribose to 5-phosphoribosyl under the catalysis of ribokinase, and further phosphorylates 5-phosphoribosyl to PRPP under the catalysis of phosphoribosyl pyrophosphatase, which serves as a substrate for hypoxanthine-guanine phosphoribosyl transferase, and condenses with NAM to produce NMN, the reaction formula is as follows:
5. the method of claim 1 or 2, wherein the enzyme catalytic system comprises hypoxanthine-guanineA kinase enzyme (MTX) ribose kinase phosphoribosyl pyrophosphate kinase, substrate and cofactor Mg 2+ The hypoxanthine-guanine phosphoribosyl transferase, ribose kinase and phosphoribosyl pyrophosphatase in the enzyme catalysis system are all purified to be free pure enzyme.
6. The method according to claim 5, wherein the nucleotide sequence encoding ribokinase is shown in SEQ.No.3 and the amino acid sequence thereof is shown in SEQ.No.4.
7. The method according to claim 5, wherein the nucleotide sequence encoding phosphoribosyl pyrophosphate kinase is shown in SEQ.No.5 and the amino acid sequence thereof is shown in SEQ.No.6.
8. The method for synthesizing beta-nicotinamide mononucleotide using an inosine-guanine phosphoribosyl transferase according to claim 1 or 2, wherein the amount of inosine-guanine phosphoribosyl transferase added is 0.1-5.0mg/ml, the amount of ribokinase added is 0.05-3.0mg/ml, and the amount of phosphoribosyl pyrophosphatase added is 0.1-6.0mg/ml in the enzyme catalyst system.
9. The method for synthesizing beta-nicotinamide mononucleotide by using hypoxanthine-guanine phosphoribosyl transferase according to claim 1 or 2, wherein the addition amount of substrate nicotinamide is 0.5-30mM and the addition amount of 5-phosphoribosyl-1-pyrophosphate is 0.75-45mM in the enzyme catalytic system; cofactor Mg 2+ The addition amount of the substrate ATP in the PRPP synthesis system is 0.1-20mM, and the addition amount of the substrate ATP in the PRPP synthesis system is 1.0-30mM; the addition amount of the D-ribose is 0.5-10mM, the reaction temperature is 25-37 ℃ and the reaction time is 2-20h.
10. A method for synthesizing β -nicotinamide mononucleotide using hypoxanthine-guanine phosphoribosyl transferase as set forth in claim 1 or 2, wherein the pH of the reaction is controlled to be 6-9.
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