Recombinant vector, engineering bacterium for synthesizing 1, 5-pentanediol and application of engineering bacterium

Abstract

The invention relates to the technical field of genetic engineering, in particular to a recombinant vector, an engineering bacterium for synthesizing 1, 5-pentanediol and application of the engineering bacterium. The recombinant vector provided by the invention can be used for expressing and synthesizing 5-amino valeraminase, lysine 2-monooxygenase, carboxylic acid reductase and phosphovinyltransferase in thalli, and recombinant bacteria constructed by utilizing the recombinant vector provided by the invention have the advantages that the recombinant bacteria can be used for synthesizing 5-amino valeraminase, lysine 2-monooxygenase, carboxylic acid reductase and phosphovinyltransferase; a complete 1, 5-pentanediol metabolic pathway can be constructed only through three key enzymes, namely 5-amino pentanamidase, lysine 2-monooxygenase and carboxylic acid reductase, the metabolic pathway is extremely short, the metabolic efficiency is higher, 1.28 g / L of 1, 5-pentanediol can be biologically synthesized under the condition that 19.43 g / L of glucose is consumed, and the strain is high in stability and suitable for industrial production.

Description

Technical Field

[0001] The present invention relates to the field of gene engineering technology, and in particular to a recombinant vector, an engineering bacterium for synthesizing 1,5-pentanediol and applications thereof. Background Art

[0002] 1,5-Pentanediol (1,5-PDO) is a diol with important industrial application value, which is widely used in the manufacture of polyesters, polyurethanes, plasticizers, inks and coatings. The biosynthesis of 1,5-pentanediol generally uses glucose as a substrate, and finally obtains 1,5-pentanediol through lysine, 5-aminovaleric acid, 5-hydroxyvaleric acid, and 5-hydroxyvaleraldehyde. However, in the process from lysine to 1,5-PDO, there are limiting factors such as long metabolic pathways and low enzyme activity. At the same time, there is also the problem of low 1,5-PDO production, which seriously restricts its application and development.

[0003] For example, Chinese patent document CN112708588A uses a recombinant microorganism that overexpresses the genes of module A, module B, and module C to complete the steps. Module A includes 4-hydroxybutyrate coenzyme A transferase abfT Aldehyde dehydrogenase bld Gene; module B includes amidase davA gene, lysine monooxygenase davB Gene, 4-aminobutyrate aminotransferase gabT Alcohol dehydrogenase yqhD Gene; Module C includes aspartate kinase lysC Gene, dihydrodipicolinate synthase dapA Diaminopimelate ddh gene; the final constructed Escherichia coli produced 1,5-pentanediol at a yield of 1.1 g / L.

[0004] Chinese patent document CN117025497A discloses another genetically engineered bacterium for catalyzing the preparation of 1,5-pentanediol. The host is Escherichia coli BL21 (DE3), which overexpresses lysine decarboxylase CadA, 4-aminobutyrate aminotransferase GabT and aldehyde-keto reductase YahK. However, the yield of 1,5-pentanediol biosynthesized by this genetically engineered bacterium is only 0.36 g / L. Summary of the invention

[0005] In order to solve the above problems, the present invention provides a recombinant vector, an engineered bacterium for synthesizing 1,5-pentanediol and its application. The recombinant bacterium constructed by using the recombinant vector provided by the present invention can realize the synthesis of 1,5-pentanediol, has an extremely short metabolic pathway, is more efficient in metabolic efficiency, has strong strain stability, and is suitable for industrial production.

[0006] In order to achieve the above object, the present invention provides the following technical solutions: The present invention provides a recombinant vector for constructing an engineering bacterium, comprising a first recombinant vector and a second recombinant vector; The first recombinant vector comprises a first starting vector and a first coding gene inserted into the first starting vector; the first coding gene comprises a 5-aminovaleramidase coding gene and a lysine 2-monooxygenase coding gene; the amino acid sequence of the 5-aminovaleramidase is shown in SEQ ID NO.28; the amino acid sequence of the lysine 2-monooxygenase is shown in SEQ ID NO.29; The second recombinant vector includes a second starting vector and a second coding gene inserted into the second starting vector; the second coding gene includes a carboxylic acid reductase coding gene and a phosphopantetheinyl transferase coding gene.

[0007] Preferably, the nucleotide sequence of the gene encoding 5-aminovaleramidase is shown as SEQ ID NO.2; the nucleotide sequence of the gene encoding lysine 2-monooxygenase is shown as SEQ ID NO.1.

[0008] Preferably, the second recombinant vector is a recombinant vector pET28a- caR - Sfp .

[0009] Preferably, the first starting vector is a pETDuet-1 vector.

[0010] Preferably, the promoter of the first coding gene is the trc promoter.

[0011] The present invention provides the use of the recombinant vector described in the above technical solution in constructing an engineering bacterium for synthesizing 1,5-pentanediol.

[0012] The present invention provides an engineering bacterium for synthesizing 1,5-pentanediol, wherein the engineering bacterium comprises an original strain and a recombinant vector transferred into the original strain; the recombinant vector is the recombinant vector described in the above technical solution; the original strain comprises Escherichia coli ( Escherichia coli ).

[0013] The present invention provides the use of the recombinant vector described in the above technical solution or the engineering bacteria described in the above technical solution in synthesizing 1,5-pentanediol.

[0014] The present invention provides a method for synthesizing 1,5-pentanediol, comprising the following steps: The engineered bacteria described in the above technical solution are fermented in a fermentation liquid containing glucose to obtain a fermentation liquid containing 1,5-pentanediol.

[0015] Preferably, the fermentation broth further contains pyruvic acid and isopropyl-β-D-thiogalactopyranoside.

[0016] Beneficial effects: The recombinant vector provided by the present invention can express and synthesize 5-aminopentanamidase, lysine 2-monooxygenase, carboxylic acid reductase and phosphopantetheinyl transferase in bacteria. The recombinant bacteria constructed by using the recombinant vector provided by the present invention can construct a complete 1,5-pentanediol metabolic pathway only through three key enzymes, 5-aminopentanamidase, lysine 2-monooxygenase and carboxylic acid reductase. The metabolic pathway is extremely short and the metabolic efficiency is more efficient. 1.28 g / L of 1,5-pentanediol can be biosynthesized under the condition of consuming 19.43 g / L of glucose. The strain has strong stability and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0018] Figure 1 for DavA Genes and DavB Electropherogram of the gene; Figure 2 for DavA Gene, caR Genes and SfP Electropherogram of the gene; Figure 3 The yields of 1,5-pentanediol, 5-hydroxyvaleric acid and 5-hydroxyvaleraldehyde in the fermentation broth of the recombinant bacterium QDE01 at different times; Figure 4 The yield of 1,5-pentanediol in the fermentation broth of recombinant bacteria QDE01, QDE02, and QDE03 at different times; Figure 5 It is the bacterial concentration of the fermentation broth of recombinant bacteria QDE01, QDE02 and QDE03 at different times. DETAILED DESCRIPTION

[0019] The present invention provides a recombinant vector for constructing an engineering bacterium, comprising a first recombinant vector and a second recombinant vector; The first recombinant vector comprises a first starting vector and a first coding gene inserted into the first starting vector; the first coding gene comprises a 5-aminovaleramidase coding gene ( DavA gene) and the lysine 2-monooxygenase encoding gene ( DavB gene); the amino acid sequence of the 5-aminovaleramidase is shown in SEQ ID NO.28; the amino acid sequence of the lysine 2-monooxygenase is shown in SEQ ID NO.29; The second recombinant vector comprises a second starting vector and a second coding gene inserted into the second starting vector; the second coding gene comprises a carboxylic acid reductase coding gene ( caRgene) and the gene encoding phosphopantetheinyltransferase ( SfP Gene). Phosphopantetheinyltransferase can phosphopantetheinylate carboxylic acid reductase, thereby increasing the specific activity of carboxylic acid reductase.

[0020] SEQ ID NO.28: MRIALYQGAPKPLDVPGNLQRLRHQAQLAAERGAQLLVCPEMFLTGYNIGLAQVERLAEAADGPAAMTVVEIAQAHRIAIVYGYPERGDDGAIYNSVQLIDAHGRSLSNYRKTHLFGELDRSMFSPGADHFPV VELEGWKVGLLICYDIEFPENARRLALDGAELILVPTANMTPYDFTCQVTVRARAQENQCYLVYANYCGAEDEIEYCGQSSIIGPDGSLLAMAGRDECQLLAELEHERVVQGRTAFPYLTDLRQELHLRKG*; SEQ ID NO.29: *.

[0021] As an embodiment, the nucleotide sequence of the gene encoding 5-aminovaleramidase is shown as SEQ ID NO.2; the nucleotide sequence of the gene encoding lysine 2-monooxygenase is shown as SEQ ID NO.1.

[0022] As an embodiment, the second recombinant vector is a recombinant vector pGCS. The recombinant vector pGCS of the present invention is a pET28a plasmid in which the T7 promoter is replaced by the trc promoter and then inserted into caR Genes and sfP The gene was constructed and published in the literature [Chen Y, Song W, Wang G, et al. Metabolic engineering of high L-lysine-producing Escherichia coli for de novo production of L-lysine-derived compounds[J]. ACS Synthetic Biology, 2024, 13(9): 2948-2959.].

[0023] As an embodiment, the first starting vector is a pETDuet-1 vector. As another embodiment, the T7 promoter in the pETDuet-1 vector is replaced with a trc promoter as the promoter of the first coding gene. As an embodiment, the DavB The gene is located between the SalⅠ and AflⅡ restriction sites of the pETDuet-1 vector. DavA The gene is located between the SacⅠ and BamHI restriction sites of the pETDuet-1 vector. The trc promoter can better express the first coding gene and the second coding gene in the host bacteria.

[0024] The recombinant vector provided by the present invention can express and synthesize 5-aminopentanamidase, lysine 2-monooxygenase, carboxylic acid reductase and phosphopantetheinyl transferase in the bacteria, and a new metabolic pathway for producing 1,5-pentanediol can be constructed by only three key enzymes, 5-aminopentanamidase, lysine 2-monooxygenase and carboxylic acid reductase. The recombinant bacteria constructed by the recombinant vector provided by the present invention can realize the synthesis of 1,5-pentanediol, the metabolic pathway is extremely short, the metabolic efficiency is more efficient, the strain stability is strong, and it is suitable for industrial production.

[0025] Based on the above advantages, the present invention provides the use of the recombinant vector described in the above technical solution in constructing an engineering bacterium for synthesizing 1,5-pentanediol.

[0026] Based on the above advantages, the present invention provides an engineering bacterium for synthesizing 1,5-pentanediol, wherein the engineering bacterium comprises an original strain and a recombinant vector transferred into the original strain; the recombinant vector is the recombinant vector described in the above technical solution; the original strain comprises Escherichia coli ( Escherichia coli ).

[0027] As an embodiment, the original strain may be Escherichia coli. As another embodiment, the Escherichia coli may be E.coli QDE. The present invention E.coli QDE is deposited in the laboratory of Qilu University of Technology and published in the literature [LiN, Xue L, Wang Z, et al. Improving the synthesis efficiency of amino acidssuch as L-lysine by assembling artificial cellulosome elements Dockerinprotein in vivo[J]. Fermentation, 2022, 8(11): 578.].

[0028] Based on the above advantages, the present invention provides the use of the recombinant vector described in the above technical solution or the engineering bacteria described in the above technical solution in the synthesis of 1,5-pentanediol.

[0029] Based on the above advantages, the present invention provides a method for synthesizing 1,5-pentanediol, comprising the following steps: The engineered bacteria described in the above technical solution are fermented in a fermentation liquid containing glucose to obtain a fermentation liquid containing 1,5-pentanediol.

[0030] As an embodiment, the fermentation broth further contains pyruvic acid and isopropyl-β-D-thiogalactopyranoside.

[0031] In order to further illustrate the present invention, the recombinant vector, the engineered bacteria for synthesizing 1,5-pentanediol and their applications provided by the present invention are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0032] Example 1 Construction of recombinant plasmid 1. Obtain from NCBI DavA Gene (GenBank: CP061335.1) and DavB The nucleotide sequence of the gene (GenBank: CP101700.1) was optimized according to the frequency of rare codon usage in Escherichia coli and commissioned to be synthesized by GenScript Biotech Co., Ltd. DavBGene sequence (SEQ ID NO.1) and DavA The gene sequence (SEQ ID NO.2) is as follows: DavB Gene (SEQ ID NO.1): DavA Gene (SEQ ID NO.2): 5'-atgcgtatcgcgctgtaccagggcgcgccgaaaccgctggatgttccgggtaacctgcagcgtctgcgtcaccaggcgcagctggcggctgaacgtggtgcgcagctgctggtgtgcccggaaatgttcctgaccggctacaacatcggcctggcgcaggttgaacgtctggcggaagcggcggacggtccggcggcgatgaccgttgttgaaatcgcgcaggcgcaccgtatcgcgatcgtttacggttacccggaacgtggtgatgatggtgcgatctacaacagcgttcagctgatcgatgcgcacggtcgttctctgtctaactaccgtaaaacccacctgttcggtgaactggatcgtagcatgttctctccgggcgcggatcacttcccggttgttgaactggaaggttggaaagttggcctgctgatctgctacgacatcgaatttccggaaaacgcgcgtcgtctggcgctggatggtgcggaactgatcctggttccgaccgcgaacatgactccgtacgatttcacctgccaggttaccgttcgtgctcgtgcgcaggaaaaccagtgctacctggtttacgcgaactactgcggtgcggaagatgaaatcgaatactgcggccagagcagcatcatcggtccggatggtagcctgctggcgatggcgggccgtgatgaatgccagctgctggcggaactggaacacgaacgtgttgttcagggccgtaccgcgttcccgtacctgaccgatctgcgtcaggaactgcacctgcgtaaaggctaa-3'.

[0033] 2. Using the optimized DavB gene sequence as a template, perform PCR amplification using primers DavB-F and DavB-R to obtain a DavB gene sequence containing the restriction site sequence; using the optimized DavAThe gene sequence was used as a template for PCR amplification using primers DavA-F and DavA-R to obtain a sequence containing restriction site. DavA Gene sequence; primer sequences are as follows: DavB-F (SEQ ID NO.3): 5'-ggatcttccagagatggatccATGAATAAAAAGAACAGGCACCCC-3'; DavB-R (SEQ ID NO.4): 5'-ctgccgttcgacgatgagctcTTAGTCGGCTAAGGCAATCGG-3'; DavA-F (SEQ ID NO.5): 5'-ggatcttccagagatgtcgacAAGGAGATATACCATGCGTATCGCGCTGTACCAG-3'; DavA-R (SEQ ID NO.6): 5'-ctgccgttcgacgatcttaagTTAGCCTTTACGCAGGTGCAGTTC-3'; Among them, 5'-ggatcttccagagatggatcc-3' (SEQ ID NO.7) is the BamHI restriction site sequence and the vector homology arm sequence; 5'-ctgccgttcgacgatgagctc-3' (SEQ ID NO.8) is the SacⅠ restriction site sequence and the vector homology arm sequence; 5'-ggatcttccagagatgtcgac-3' (SEQ ID NO.9) is the SalⅠ restriction site sequence and the vector homology arm sequence; 5'-ctgccgttcgacgatcttaag-3' (SEQ ID NO.10) is the AflⅡ restriction site sequence and the vector homology arm sequence; 5'-AAGGAGATATACC-3' (SEQ ID NO. 35) is the ribosome binding site (RBS) sequence.

[0034] The reaction system for PCR amplification was as follows: 2 μL template, 2 μL upstream primer, 2 μL downstream primer, 25 μL 2×Phanta ultra-high fidelity enzyme and ddH 2 O 19 μL. The reaction program of PCR amplification was as follows: 95°C pre-denaturation for 3 min; 95°C denaturation for 15 s, 60°C annealing for 15 s, 72°C extension for 45 s, 30 cycles; 72°C final extension for 10 min.

[0035] 3. Entrust Sangon Biotechnology Co., Ltd. to synthesize the trc promoter sequence (shown in SEQ ID NO.11), as follows: 5'--3'; The trc promoter sequence was connected between the SphI and NdeI restriction sites of the pETDuet-1 vector to replace the T7 promoter on the pETDuet-1 vector, thereby obtaining a new pETDuet-1 vector (referred to as the pETDuet vector).

[0036] 4. Combine the pETDuet vector obtained in step 3 and the vector obtained by amplification in step 2 DavB The gene sequence was treated with SalⅠ and AflⅡ respectively, and then connected at 37℃ overnight to obtain the recombinant plasmid pETDuet-d avB .

[0037] 5. The recombinant plasmid pETDuet- davB and the amplified one in step 2 DavBThe gene sequences were treated with SacⅠ and BamHI respectively, and then connected at 37℃ overnight to obtain the recombinant plasmid pETDuet- davB - davA .

[0038] Example 2 Construction of Escherichia coli for the synthesis of 1,5-pentanediol Plasmid pET28a- caR - sfP The recombinant plasmid pETDuet- davB - davA Transformed to the same strain of E. coli E.coli In QDE, the cells were cultured overnight at 37°C in LB medium containing 50 µg / mL kanamycin and 100 µg / mL ampicillin for dual resistance.

[0039] Single colonies were selected for PCR identification (primers see Table 1). The results were as follows: Figure 1 and Figure 2 As shown, Figure 1 M stands for Marker DL2000, 1 stands for DavA Gene, 2 DavB Gene; Figure 2 M stands for Marker DL5000, 1 stands for DavA Gene, 2 caR Gene, 3 SfP Gene. Determine DavB Gene, DavA Gene, caR Gene, Sfp The gene was successfully transferred into Escherichia coli to obtain the recombinant bacterium QDE01.

[0040] Table 1 Primer sequences used for PCR identification

[0041] Example 3 Biosynthesis of 1,5-pentanediol by recombinant bacterium QDE01 The recombinant bacteria QDE01 constructed in Example 2 was activated in a shaker at 37°C. 600 The seed solution was inoculated into LB medium containing 0.5 g / L pyruvic acid at a 1% (v / v) inoculum and cultured on a shaking platform until OD 600 When the value was 1, isopropyl-β-D-thiogalactopyranoside (IPTG) with a final concentration of 0.02 mmol / L and 20 g / L glucose were added, and the fermentation was carried out at 30 °C with shaking and regular sampling.

[0042] Set up a control group: replace the recombinant bacteria QDE01 with a control strain for fermentation; the construction method of the control strain is similar to that of the recombinant bacteria QDE01, except that the plasmid pET28a- caR - sfPreplaced with plasmid pET28a (T7 promoter replaced by trc promoter), and recombinant plasmid pETDuet- davB - davA Replace with the pETDuet vector constructed in Example 1.

[0043] The production of 1,5-pentanediol, 5-hydroxyvaleric acid and 5-hydroxyvaleraldehyde in the fermentation broth was detected every 1 day by ultraviolet high performance liquid chromatography. Figure 3 shown.

[0044] Depend on Figure 3 It can be seen that the present invention expresses the optimized codon DavB Gene 、DavA Genes and caR Genes, using 5-aminopentylamidase, lysine 2-monooxygenase and carboxylic acid reductase three key enzymes can construct a complete 1,5-pentanediol metabolic pathway, which can biosynthesize 1.28g / L of 1,5-pentanediol under the condition of consuming 19.43g / L glucose. A complete and efficient 1,5-pentanediol biosynthetic pathway was constructed. E.coli QDE did not produce 1,5-pentanediol.

[0045] Comparative Example 1 The construction method of recombinant bacterium QDE02 is similar to that of recombinant bacterium QDE01, except that the recombinant plasmid pETDuet- davB - davA Replaced with recombinant plasmid pETDuet- davB - davA -g abT - yahK ; The construction method of recombinant bacterium QDE03 is similar to that of recombinant bacterium QDE01, except that the recombinant plasmid pETDuet- davB - davA Replaced with recombinant plasmid pETDuet- davB - davA - davT - eyqhD .

[0046] The recombinant plasmid pETDuet- davB - davA -g abT - yahK The construction method is as follows: 1) Containing the gene encoding 4-aminobutyrate aminotransferase ( gabT The plasmid pGGY containing the gene was used as a template and PCR amplification was performed using primers gabT-F and gabT-R to obtain gabT Gene sequence; yahK The plasmid pGGY of the gene was used as a template and PCR amplification was performed using primers yahK-F and yahK-R to obtain yahK Gene sequence; 2) The gabT The gene sequence was inserted into the recombinant plasmid pETDuet- davB - davAThe recombinant plasmid pETDuet- davB - davA -g abT ; 3) yahK The gene sequence was inserted into the recombinant plasmid pETDuet- davB - davA -g abT The recombinant plasmid pETDuet- davB - davA -g abT - yahK .

[0047] The recombinant plasmid pETDuet- davB - davA - davT - eyqhD The construction method is as follows: 1) Containing davT The plasmid pGDE containing the gene was used as a template and PCR amplification was performed using primers davT-F and davT-R to obtain davT Gene sequence; eyqhD The plasmid pGDE containing the gene was used as a template and PCR amplification was performed using primers eyqhD-F and eyqhD-R to obtain eyqhD Gene sequence; 2) The davT The gene sequence was inserted into the recombinant plasmid pETDuet- davB - davA The recombinant plasmid pETDuet- davB - davA - davT ; 3) eyqhD The gene sequence was inserted into the recombinant plasmid pETDuet- davB - davA - davT The recombinant plasmid pETDuet- davB - davA - davT - eyqhD .

[0048] The plasmid pGGY and plasmid pGDE are both disclosed in the literature [Chen Y, Song W, Wang G, et al. Metabolic engineering of high L-lysine-producing Escherichia coli for de novoproduction of L-lysine-derived compounds [J]. ACS Synthetic Biology, 2024, 13(9): 2948-2959.]. The primer sequences are shown in Table 2.

[0049] Table 2 Sequences of different primers

[0050] Note: 5'-taagaaggagatatacatatg-3' (SEQ ID NO.30) is the NdeⅠ restriction site sequence and the vector homology arm sequence; 5'-tggccggccgatatccaattg-3' (SEQ ID NO.31) is the MunⅠ restriction site sequence and the vector homology arm sequence; 5'-atcggccggccacgcgatcgcaaggag-3' (SEQ ID NO.32) is the SgfⅠ restriction site sequence and the vector homology arm sequence; 5'-ctgccgttcgacgatggtacc-3' (SEQ ID NO.33) and 5'-tttrccagactcgagggtrcc-3' (SEQ ID NO.34) are both KpnⅠ restriction site sequences and vector homology arm sequences.

[0051] Comparative Example 2 After the constructed recombinant bacteria QDE01, QDE02, and QDE03 were verified to be correct, they were fermented and sampled at regular intervals according to the method of Example 3. The yield and bacterial concentration of 1,5-pentanediol in the fermentation broth of different recombinant bacteria were detected. The results are shown in Figure 4 and Figure 5 .

[0052] The results show that the 1,5-PDO production of the recombinant bacterium QDE01 reached 1.3 g / L at 6 days. The 1,5-PDO production of strains QDE02 and QDE03 was only 1.13 g / L and 0.87 g / L at 6 days. In addition, the bacterial concentrations of QDE02 and QDE03 were significantly lower than that of the recombinant bacterium QDE01. It can be seen that too many genes inserted into the plasmid are not conducive to the growth of the recombinant bacteria, thereby affecting the production of 1,5-pentanediol. The recombinant bacterium QDE01 constructed by the present invention has a shorter metabolic pathway, high 1,5-pentanediol production, and good bacterial growth, which is more conducive to later industrial production.

[0053] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. Construct a recombinant vector of an engineering bacterium, characterized in that: comprising a first recombinant vector and a second recombinant vector; The first recombinant vector comprises a first starting vector and a first coding gene inserted into the first starting vector; the first coding gene comprises a 5-aminovaleramidase coding gene and a lysine 2-monooxygenase coding gene; the amino acid sequence of the 5-aminovaleramidase is shown in SEQ ID NO.28; the amino acid sequence of the lysine 2-monooxygenase is shown in SEQ ID NO.29; The second recombinant vector includes a second starting vector and a second coding gene inserted into the second starting vector; the second coding gene includes a carboxylic acid reductase coding gene and a phosphopantetheinyl transferase coding gene.

2. The recombinant vector according to claim 1, characterized in that The nucleotide sequence of the gene encoding 5-aminovaleramidase is shown in SEQ ID NO.2; the nucleotide sequence of the gene encoding lysine 2-monooxygenase is shown in SEQ ID NO.

1.

3. The recombinant vector according to claim 1, characterized in that The second recombinant vector is the recombinant vector pET28a- R - Sf .

4. The recombinant vector according to claim 1, characterized in that The first starting vector is a pETDuet-1 vector.

5. The recombinant vector according to claim 1, characterized in that The promoter of the first coding gene is trc promoter.

6. Use of the recombinant vector according to any one of claims 1 to 5 in constructing an engineered bacterium for synthesizing 1,5-pentanediol.

7. An engineered bacterium for synthesizing 1,5-pentanediol, characterized in that: The engineered bacteria include an original strain and a recombinant vector transferred into the original strain; the recombinant vector is the recombinant vector according to any one of claims 1 to 5; the original strain includes Escherichia coli ( Escherichia coli ).

8. Use of the recombinant vector according to any one of claims 1 to 5 or the engineered bacteria according to claim 7 in the synthesis of 1,5-pentanediol.

9. A method for synthesizing 1,5-pentanediol, characterized in that: The following steps are involved: The engineered bacteria according to claim 7 are fermented in a fermentation liquid containing glucose to obtain a fermentation liquid containing 1,5-pentanediol.

10. The method according to claim 9, characterized in that The fermentation broth also contains pyruvic acid and isopropyl-β-D-thiogalactopyranoside.

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