CN110564757A - Construction method and application of metabolic engineering escherichia coli strain for producing 3-hydroxypropionic acid by using acetic acid or salt thereof - Google Patents
Construction method and application of metabolic engineering escherichia coli strain for producing 3-hydroxypropionic acid by using acetic acid or salt thereof Download PDFInfo
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Abstract
The invention provides a construction method of a metabolic engineering escherichia coli strain for producing 3-hydroxypropionic acid by using acetic acid or salts thereof, which is further modified on the basis of a metabolic pathway for producing 3-HP by using the acetic acid or the salts thereof through acetyl-CoA carboxylase and malonyl-CoA reductase and comprises one or two of the following two pathways: (1) inhibit fatty acid synthesis and promote the glyoxylate cycle to increase the metabolic flux of acetyl-CoA and malonyl-CoA to the target metabolites and enhance energy metabolism; (2) fatty acid synthesis pathways are inhibited to increase the metabolic flux of acetyl-CoA and malonyl-CoA to the metabolites of interest. According to the invention, on the basis of the established exogenous expression 3-HP pathway, metabolic pathway and regulation are analyzed, escherichia coli is modified by using a genetic engineering means, and the obtained strain produces 3-HP in a culture medium taking acetic acid or salts thereof as a carbon source, wherein the yield can reach 0.38g/g and is 50.7% of the maximum theoretical yield.
Description
Technical Field
The invention belongs to the technical field of bioengineering, and particularly relates to a construction method and application of a metabolic engineering escherichia coli strain for producing 3-hydroxypropionic acid by using acetic acid or salts thereof.
Background
Acetic acid is a cheap and readily available carbon source and is currently synthesized predominantly by chemical methods (Li et al, 2016). The synthesis process is mainly a methanol carbonylation method, and the yield of acetic acid synthesized by the method exceeds 64 percent of the total yield of acetic acid worldwide. Acetic acid can be produced not only by chemical industry, but also in large quantities in nature, and microbial fermentation, lignocellulose hydrolysis and anaerobic microbial anaerobic digestion all produce excessive waste acetic acid. In addition, acetic acid can also be biologically derived from CO2、CO、CH4Equal to one carbon compound (Hu et al, 2016; Kalyuzhnaya et al, 2013; Straub et al, 2014).
3-hydroxypropionic acid (3-HP), a three-carbon achiral organic acid, is an isomer of 3-hydroxypropionic acid and lactic acid (2-hydroxypropionic acid), and thus 3-hydroxypropionic acid is also called hydroxypropionic acid or ethylene lactic acid. The U.S. department of energy (DOE) 2004 proposed 12 important platform compounds, 3-hydroxypropionic acid ranked in the fourth place. Due to its excellent chemical properties, 3-hydroxypropionic acid can be used to produce a wide variety of chemical derivatives, such as acrylic acid, 1, 3-propanediol, methacrylates, acrylamide, malonic acid, and the like.
currently, the main methods for producing 3-hydroxypropionic acid are still chemical synthesis methods such as the oxidation of bromo 3-hydroxypropanal, the hydrolysis of propionic acid, and the 2-cyanoglycolic acid-base method, which cause many problems of environmental pollution. Recently, much interest has been focused on the biochemical production of 3-hydroxypropionic acid from renewable resources to meet the demand for environmental and economic improvements. To produce 3-hydroxypropionic acid using metabolic engineering, a number of biosynthetic pathways have been constructed that use glucose or glycerol as starting materials (Jiang et al, 2009; Huang et al, 2013). Recently, several engineered E.coli strains harboring coenzyme B12-dependent glycerol dehydratase (GDHt) and aldehyde dehydrogenase (ALDH) were developed for the production of 3-hydroxypropionic acid using glycerol (Raj et al, 2008; Jo et al, 2008; Kim et al, 2014). Since Escherichia coli cannot produce vitamin B12, it is necessary to supply coenzyme B12 at high cost from an external source. However, the production of 3-hydroxypropionic acid using acetic acid or a salt thereof as a carbon source has been reported. Ji HL et al synthesized 3-hydroxypropionic acid using acetic acid, and increased accumulation of 3-hydroxypropionic acid by inhibiting fatty acid synthesis and promoting glyoxylate cycle by adding Cerulenin and knocking out iclR, respectively.
Chinese patent application 201710969403.0 discloses a construction method and application of metabolic engineering Escherichia coli strain for producing hydroxypropionic acid by using acetic acid, wherein wild Escherichia coli is used as starting bacteria to construct 3-hydroxypropionic acid and lactic acid and a synthetic way for co-producing 3-hydroxypropionic acid and lactic acid. The acetyl CoA is used as a key precursor substance of the hydroxypropionic acid and is also a key node of central metabolism, so that the metabolic flux of the acetyl CoA flowing to a target product can be increased by regulating the metabolic flux of the acetyl CoA by down-regulating gltA, the yield of the hydroxypropionic acid can be improved, the central metabolism can be maintained, the supply of the precursor substance necessary for energy and growth is ensured, and the yield of the 3-hydroxypropionic acid is lower.
Disclosure of Invention
The first purpose of the invention is to provide a construction method of metabolic engineering Escherichia coli strain for producing 3-hydroxypropionic acid by using acetic acid or salt thereof with higher yield.
The second purpose of the invention is to provide a metabolic engineering Escherichia coli strain obtained by the construction method.
The third purpose of the invention is to provide the application of the metabolic engineering Escherichia coli strain obtained by the construction method in the fermentation production of 3-hydroxypropionic acid by taking acetic acid or salts thereof as a carbon source.
In order to achieve the first object, the invention discloses the following technical scheme: a method of constructing a metabolically engineered escherichia coli strain that utilizes acetic acid or a salt thereof to produce 3-hydroxypropionic acid, the method further engineered on the basis of a metabolic pathway for producing 3-hydroxypropionic acid utilizing acetic acid or a salt thereof via an acetyl-CoA carboxylase and a malonyl-CoA reductase, the further engineering comprising one or both of: (1) inhibit fatty acid synthesis and promote the glyoxylate cycle to increase the metabolic flux of acetyl-CoA and malonyl-CoA to the target metabolites and enhance energy metabolism; (2) fatty acid synthesis pathways are inhibited to increase the metabolic flux of acetyl-CoA and malonyl-CoA to the metabolites of interest.
As a preferable mode, the metabolic pathway for producing 3-hydroxypropionic acid using acetic acid or a salt thereof via acetyl-CoA carboxylase and malonyl-CoA reductase means that an acetyl-CoA carboxylase-encoding gene (dtsR1 and accBC) derived from Corynebacterium glutamicum and a malonyl-CoA reductase-encoding gene (mcr) derived from filamentous fungus Lloyothiacola virens (C.aurantiacaa) are overexpressed.
Among them, the protein expressed by dtsR1 is a subunit of acetyl-CoA carboxylase (Acc) and is a component of the Acc gene, and this gene is included in the whole Acc gene.
As a preferred embodiment, acetic acid or a salt thereof refers to acetic acid or an acetate salt including one or both of sodium acetate and ammonium acetate. Acetate can also be used as a carbon source to achieve consistent results, and when acetic acid is added to the fermentation medium, the pH will decrease and need to be adjusted to a neutral pH with sodium hydroxide, and acetate will be produced upon neutralization of the acetic acid. Acetate such as sodium acetate and ammonium acetate may be directly placed in the medium, and acetate may be used as a carbon source.
As a preferred embodiment, the further modification comprises one or both of the following two approaches:
(1) Knocking out fadR by host bacteria;
(2) fabR is overexpressed.
Preferably, the further modification refers to knocking out fadR and over-expressing fabR by the host bacteria.
Preferably, the further modification refers to knocking out fadR or over-expressing fabR of the host bacterium.
In order to achieve the second object, the invention discloses the following technical scheme: the metabolic engineering Escherichia coli strain obtained by the construction method.
In order to achieve the third object, the invention discloses the following technical scheme: the metabolic engineering escherichia coli strain obtained by the construction method is applied to the fermentation production of 3-hydroxypropionic acid by taking acetic acid or salts thereof as a carbon source.
The metabolically engineered Escherichia coli uses acetic acid or salts thereof as raw materials to produce 3-hydroxypropionic acid by fermentation, is mainly established on the basis of cloning encoding genes of key enzymes from different sources in a pathway for synthesizing the 3-hydroxypropionic acid by malonyl-CoA and constructing a complete pathway for producing the 3-hydroxypropionic acid, and combines the modification of host bacteria or the overexpression of fatty acid synthesis related enzymes, including the modification of a transport pathway of the acetic acid or the salts thereof, so as to enhance the utilization of the acetic acid or the salts thereof; in addition, by inhibiting fatty acid synthesis, the flow of acetyl-CoA and malonyl-CoA to the fatty acid synthesis pathway is reduced, thereby enhancing metabolic flux to the target product. According to the invention, through analysis of metabolic pathway and regulation and control, Escherichia coli is transformed by using a genetic engineering means, the transformation is carried out on the basis of metabolic engineering Escherichia coli capable of producing 3-hydroxypropionic acid by using acetic acid or salts thereof as a carbon source, and the 3-hydroxypropionic acid is produced by using the further transformed metabolic engineering strain and using the acetic acid or salts thereof as a main carbon source.
The method of the invention uses Red recombination technology to knock out fadR; in addition, fabR was overexpressed using molecular biology techniques.
the invention takes escherichia coli BL27 as a starting bacterium, and is further modified on the basis of the escherichia coli which has heterologously expressed key enzyme for synthesizing 3-hydroxypropionic acid by a malonyl-CoA pathway. The invention takes acetic acid or salt thereof as a carbon source, which means that the acetic acid or salt thereof is taken as a main carbon source, and malonyl-CoA is a key intermediate for producing 3-hydroxypropionic acid and can also be taken as a precursor substance for fatty acid synthesis, so that the metabolic flow of malonyl-CoA to 3-hydroxypropionic acid is increased by inhibiting a fatty acid synthesis pathway, thereby improving the yield of 3-hydroxypropionic acid. By knocking out the fadR gene, on one hand, fatty acid degradation can be inhibited, and fatty acid synthesis can be inhibited, so that the metabolic flow of malonyl-CoA to 3-hydroxypropionic acid is increased; on the other hand, indirectly promoting the glyoxylate pathway increases energy metabolism; overexpression of fabR also inhibits fatty acid synthesis and increases metabolic flux of intermediates to 3-hydroxypropionic acid.
The invention has the advantages that: according to the invention, on the basis of the established exogenous expression 3-hydroxypropionic acid pathway, metabolic pathway and regulation are analyzed, escherichia coli is modified by using a genetic engineering means, and the obtained strain produces 3-hydroxypropionic acid in a culture medium taking acetic acid or salt thereof as a carbon source, wherein the yield can reach 0.38g/g and is 50.7% of the maximum theoretical yield.
Drawings
FIG. 1 is a diagram showing the metabolism of 3-hydroxypropionic acid (3-HP) produced by E.coli using acetic acid or its salt. Acetate, Acetate salt; AcP, acetyl phosphate; acs, acetyl-CoA synthetase; ackA, acetate kinase; pta, phosphotransacetylase; acacoa, acetyl-CoA; Malonyl-CoA, Malonyl-CoA; acc, acetyl-CoA carboxylase; mcr, malonyl-CoA reductase; gltA, citrate synthase; CIT, citric acid; acnAB, aconitase; ICT, isocitric acid; icdA, isocitrate dehydrogenase; α KG, α ketoglutaric acid; sucAB, alpha ketoglutarate dehydrogenase; SucCoA, succinyl-CoA; sucCD, succinate thiokinase; SUC, succinic acid; aceA, isocitrate lyase; iclR, isocitrate lyase repressor; GOX, glyoxylic acid; aceB, malate synthase; MAL, malic acid; mdh, malate dehydrogenase; OAA, oxaloacetate; fumABC, fumarase; FUM, fumaric acid; frdABCD, fumarate reductase; sdhABCD, succinate dehydrogenase.
Detailed Description
hereinafter, the technique of the present invention will be described in detail with reference to specific embodiments. The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
Example 1.construction of the 3-hydroxypropionic acid production pathway
pET28a plasmid is used as carrier, and key enzyme of 3-hydroxy propionic acid production path from Chloroflexus aurantiacaus and Corynebacterium glutamicum is overexpressed at the same time. The enzymes that produce 3-hydroxypropionic acid from acetyl-CoA are acetyl-CoA carboxylase (encoded by dtsR1 and accBC derived from Corynebacterium glutamicum), malonyl-CoA reductase (encoded by mcr derived from Chloroflexus aurantiacaus), respectively. A recombinant plasmid pET28a-mcr-RBS-dtsR1-RBS-accBC (pET 28a-MDA for short) is obtained by expressing more than three genes derived from Chloroflexus aurantiaca and Corynebacterium glutamicum in pET28a at the same time. The pET28a vector carries a T7 promoter which is strictly controlled by T7RNA polymerase, and since wild Escherichia coli does not express T7RNA polymerase per se, it needs to be modified to synthesize T7RNA polymerase, thereby regulating the T7 expression system. Coli containing T7RNA polymerase can express target protein from T7 promoter. In addition, the pET28a vector has a segment of gene lac I coding lac repressor protein immediately downstream of the T7 promoter, which can block the transcription of target gene caused by T7RNA polymerase from any source, and an inducer (lactose or lactose analogue) is added to combine with the lac repressor protein to make it lose its effect.
The plasmid construction method is as follows:
The sequences of mcr from Chloroflexus aurantiacus, dtsR1 from Corynebacterium glutamicum and accBC gene are searched in NCBI, the three genes are expressed in series, RBS optimized by calculation is added among the genes, and finally, a linearized pET28a vector is connected.
Plasmids were constructed using a seamless cloning approach. The constructed plasmid was transformed into e.coli DH5 α by calcium transfer, and then verified by colony PCR and sequencing. The primers for amplifying the different fragments are shown in Table 1 below.
TABLE 1 overexpression genes, promoter replacement and Gene knockout primers
Example 2 construction of FadR Gene knock-out bacteria
Since acetyl-CoA and malonyl-CoA are key intermediates in the production of 3-hydroxypropionic acid and can also serve as precursor substances for fatty acid synthesis, the metabolic flux of acetyl-CoA and malonyl-CoA to 3-hydroxypropionic acid is increased by inhibiting the fatty acid synthesis pathway, thereby increasing the yield of 3-hydroxypropionic acid. FadR is a global regulator of lipid metabolism (globalregulator) and also of Fatty Acid (FA) metabolism, and can be used as catabolism inhibitor and anabolism activator. Meanwhile, FadR indirectly regulates aceBAK by activating the expression of IclR so as to inhibit the glyoxylate cycle.
Knocking out the fadR gene by adopting a Red recombination method. A strain in which fadR is knocked out on the basis of e.coli BL27 is named LNY02, and a strain in which fadR is knocked out on the basis of LNY01 (table 2) is named LNY 04. The specific operation of gene knockout is as follows:
For the fadR knock-out, primers (primer sequences shown in Table 1) were first designed and about 1700bp DNA fragment with kanamycin resistance was cloned by PCR. The fragment is transferred into the electrotransformation competence of the host bacterium through electrotransfer, and the preparation of the electrotransfer competence of the host bacterium, the verification of knockout and the elimination of kanamycin resistance are specifically operated as follows:
The host cells were transformed with calcium and introduced into plasmid pKD46, and the recombinants were screened for ampicillin. The recombinant strain introduced with pKD46 was cultured at 30 ℃ to OD600About 0.2, L-arabinose was added to the final concentration of 10mM for 1 hour of induction, and then electroporation competence was prepared using 10% glycerol. The kanamycin fragment obtained above was transferred into a prepared electrotransformation competence. Electrotransformation was carried out using bacterial mode 1(1.8KV, 5ms) and transformants which underwent homologous recombination were selected using kanamycin. Designing a replacement verification primer, verifying whether the promoter is successfully replaced by adopting a colony PCR method, and further sequencing and verifying the PCR verified positive clone to ensure that the replaced promoter region has no mutation, wherein the used primers are shown in the table 1.
And (3) replacing the successfully-replaced recombinant bacteria with the promoter, culturing at 37 ℃ for 5-6 hours, transferring to 42 ℃ for overnight culture, separating a single colony, and then verifying the resistance. Only kanamycin-resistant and ampicillin-resistant colonies were the colonies which had been eliminated with pKD 46. Then, the strain was transformed into the plasmid pCP20, and the chloramphenicol resistant plate was cultured at 30 ℃ for a certain period of time, then, it was transformed into 42 ℃ for overnight culture, and a single colony was isolated, followed by verifying the resistance. Colonies that grew only on the non-resistant plates, but not on both the kanamycin-resistant plates and the ampicillin-resistant plates were picked for identification. By using the verification primers to carry out colony PCR, the size of a transformant with successfully eliminated resistance is similar to that of a target fragment obtained by wild bacteria PCR, and the size of the transformant with successfully eliminated resistance is obviously different from that of a PCR fragment of a strain with not eliminated resistance, so that whether the elimination and recombination of the resistance gene is successful or not can be judged.
TABLE 2 strains and plasmids
Example 3 construction of fabR overexpression plasmids
The binding site of FabR overlaps with the binding site of FadR, which inhibits the expression of fabA and fabB, and the binding site of FabR is the transcriptional activator of fabA and fabB, and is located in the promoter regions of fabA and fabB. Both regulators can bind to the fabB promoter simultaneously in the presence of unsaturated acyl-ACP (Zhu et al, 2009).
The seamless cloning method is adopted to construct a plasmid pBAD33-Trc-fabR to over-express fabR. The specific operation is as follows:
The plasmid pBAD33 is used as a vector, primers are designed, an E.coli is used as a template, a fabR gene is cloned by PCR, a trc promoter is added in front of the fabR gene, and finally the linearized pBAD33 vector is connected. The subsequent operation was the same as in example 1.
Example 4 knock-out fadR Strain Shake flask fermentation
Coli BL27 or its deletion bacterium is used as the starting bacterium, and BL27(pET-28a-MDA), LNY01(pET-28a-MDA), LNY02(pET-28a-MDA) and LNY04(pET-28a-MDA) are respectively obtained by calcium transfer into the plasmid for constructing the production pathway of 3-hydroxypropionic acid. Hereinafter pET-28a-MDA is abbreviated as MDA.
And (3) shake flask fermentation operation: single colonies on the plate were picked and inoculated in a tube containing 3mL of LB medium overnight, and then transferred to a flask containing 50mL of LB medium at 1% inoculum size under conditions of 37 ℃ and 220 rpm. After 8h of secondary seed culture, the secondary seeds are transferred into a shake flask fermentation medium (M9 medium). M9 medium was supplemented with 10g/L sodium acetate, 5g/L yeast extract and 40mg/L biotin. The inoculum size was 2%, the culture conditions were 37 ℃, 220rpm, to OD600To about 1.0, the inducer IPTG was added to a final concentration of 0.1 mM. The post-induction culture conditions were 25 ℃ and 220 rpm. 3M H was used in the cultivation2SO4The pH was maintained at 7. Antibiotics (kanamycin to 50mg/L, chloramphenicol to 34mg/L) were added to the above medium as necessary.
m9 medium composition (per liter): na (Na)2HPO4·12H2O 15.12g，KH2PO4 3g，NaCl 0.5g，MgSO4·7H2O 0.5g，CaCl2 0.011g，NH4Cl 1g and 1% vitamin B1 0.2mL。
the method for measuring the 3-hydroxypropionic acid and the acetic acid comprises the following steps: during the shake flask fermentation culture, samples were taken at intervals of 8 hours, and centrifuged at 12000rpm for 10 minutes to separate the cells from the supernatant. Filtering the supernatant of the fermentation liquor by a 0.22-micron microporous membrane, and monitoring 3-hydroxypropionic acid and acetic acid organic acid in the supernatant of the fermentation liquor by using an Shimadzu high performance liquid chromatograph. The column was a BioRadaminex HPX-87 ion column (300 mm. times.7.8 mm) equipped with an ultraviolet detector and a differential refraction detector. Mobile phase 2.5mM H2SO4The flow rate was 0.5mL/min, and the column temperature was 50 ℃.
Measurement of cell concentration the absorbance at 600nm was measured by spectrophotometry.
The results of the shake flask fermentation are shown in Table 3. After acetic acid uptake is enhanced and fadR is knocked out, the yield of the strain LNY04(MDA) reaches 1.77g/L, and is improved by 14.2% compared with the yield of the original strain BL27 (MDA).
TABLE 3 yield and yield of 3-hydroxypropionic acid from genetically engineered bacteria
。
Example 5 Shake flask fermentation of over-expressing fabR Strain
Coli BL27 or its deletion bacterium is used as the starting bacterium, plasmid for constructing 3-hydracrylic acid production pathway is transferred by calcium transfer and overexpression gene plasmid is transferred to obtain BL27(pET-28a-MDA, pBAD33-Trc-fabR), LNY01(pET-28a-MDA, pBAD33-Trc-fabR), LNY02(pET-28a-MDA, pBAD33-Trc-fabR) and LNY04(pET-28a-MDA, pBAD33-Trc-fabR) respectively. Hereinafter pET-28a-MDA, pBAD33-Trc-fabR is abbreviated as MDAF.
The shake flask fermentation procedure, M9 medium composition, 3-hydroxypropionic acid and acetic acid assay and cell concentration assay were as described in example 3.
The shake flask fermentation results are shown in table 4. After the acetic acid uptake is enhanced and fabR is over-expressed, the yield of the strain LNY01(MDAF) is 1.67g/L, the yield reaches 0.38g/g, and is 50.7 percent of the maximum theoretical yield.
TABLE 4 yield and yield of 3-hydroxypropionic acid from genetically engineered bacteria
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
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<210> 10
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 10
ttcagaacca gtactaactt a 21
<210> 11
<211> 70
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 11
tctggtatga tgagtccaac tttgttttgc tgtgttatgg aaatctcact cgtcttgagc 60
gattgtgtag 70
<210> 12
<211> 70
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 12
aacaacaaaa aacccctcgt ttgaggggtt tgctctttaa acggaaggga gatgtaacgc 60
actgagaagc 70
<210> 13
<211> 30
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 13
cacaggaaac agaccatgtt cattctctgg 30
<210> 14
<211> 35
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 14
ccgccaaaac agccaagctt ttactcgtcc ttcac 35
<210> 15
<211> 34
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 15
gggctagcga attcgagctc gcgcaacgca atta 34
<210> 16
<211> 30
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 16
ccagagaatg aacatggtct gtttcctgtg 30
Claims (8)
1. A method of constructing a metabolically engineered escherichia coli strain that utilizes acetic acid or a salt thereof to produce 3-hydroxypropionic acid, wherein the metabolic pathway for producing 3-hydroxypropionic acid utilizing acetic acid or a salt thereof via acetyl-CoA carboxylase and malonyl-CoA reductase is further engineered to include one or both of: (1) inhibit fatty acid synthesis and promote the glyoxylate cycle to increase the metabolic flux of acetyl-CoA and malonyl-CoA to the target metabolites and enhance energy metabolism; (2) fatty acid synthesis pathways are inhibited to increase the metabolic flux of acetyl-CoA and malonyl-CoA to the metabolites of interest.
2. The method of claim 1, wherein the metabolic pathway for producing 3-hydroxypropionic acid using acetic acid or a salt thereof via acetyl-CoA carboxylase and malonyl-CoA reductase means that genes encoding acetyl-CoA carboxylase (dtsR1 and accBC) derived from Corynebacterium glutamicum and genes encoding malonyl-CoA reductase (mcr) derived from filamentous fungus Fusarium chlororaphiae (C.aurantiacaa) are overexpressed.
3. The method for constructing the metabolically engineered escherichia coli strain for the production of 3-hydroxypropionic acid using acetic acid or a salt thereof as set forth in claim 1, wherein the acetic acid or a salt thereof is acetic acid or an acetate salt comprising one or both of sodium acetate and ammonium acetate.
4. The method for constructing a metabolically engineered escherichia coli strain for the production of 3-hydroxypropionic acid using acetic acid or a salt thereof as claimed in claim 1, wherein said further engineering comprises one or both of the following two pathways:
(1) knocking out fadR by host bacteria;
(2) fabR is overexpressed.
5. The method for constructing a metabolically engineered escherichia coli strain for producing 3-hydroxypropionic acid using acetic acid or a salt thereof as claimed in claim 1, wherein the further modification is to knock out fadR and overexpress fabR in the host strain.
6. The method for constructing a metabolically engineered escherichia coli strain for producing 3-hydroxypropionic acid using acetic acid or a salt thereof according to claim 1, wherein the further modification is to knock out fadR or overexpress fabR in the host bacterium.
7. A metabolically engineered Escherichia coli strain obtained by the construction method according to any one of claims 1 to 6.
8. Use of a metabolically engineered E.coli strain obtained by the method of any one of claims 1 to 6 for the fermentative production of 3-hydroxypropionic acid using acetic acid or a salt thereof as a carbon source.
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