CN114774451A - Recombinant escherichia coli and method for producing tyrosol, hydroxytyrosol or salidroside by using same - Google Patents

Recombinant escherichia coli and method for producing tyrosol, hydroxytyrosol or salidroside by using same Download PDF

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CN114774451A
CN114774451A CN202210466075.3A CN202210466075A CN114774451A CN 114774451 A CN114774451 A CN 114774451A CN 202210466075 A CN202210466075 A CN 202210466075A CN 114774451 A CN114774451 A CN 114774451A
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tyrosol
salidroside
hydroxytyrosol
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刘建忠
孙华为
叶迪凯
王佑琪
田嘉源
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Sun Yat Sen University
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Abstract

The invention discloses a recombinant escherichia coli and a method for producing tyrosol, hydroxytyrosol or salidroside by the recombinant escherichia coli, wherein the recombinant escherichia coli expresses tyrosol synthesis pathway enzyme fusion in host bacteria to produce tyrosol, then introduces 4-hydroxyphenylacetic acid 3-hydroxylase to produce hydroxytyrosol, and introduces glycosyltransferase to produce salidroside; spy (or Snoop) cyclization can also be treated by heat to increase cell permeability, avoiding mass transport limitations of the cells. The long metabolic pathway is divided into a plurality of pathways through the module co-culture, and different host cells can be used for expression, so that the metabolic burden on the host cells is reduced; moreover, different routes can be selected from the most suitable host cells for expression; the adaptability of the module can be controlled by adjusting the proportion of the module bacteria; plug and play, easy to assemble into different pathways to synthesize different products. And can also avoid the toxic influence of metabolic intermediates and target products.

Description

Recombinant escherichia coli and method for producing tyrosol, hydroxytyrosol or salidroside by using same
Technical Field
The invention belongs to the technical field of bioengineering, and particularly relates to recombinant escherichia coli and a method for producing tyrosol, hydroxytyrosol or salidroside by using the recombinant escherichia coli.
Background
Tyrosol, p-hydroxyphenylethanol, is a natural phenolic antioxidant. It has antioxidant, antifatigue, antihypoxic, anti-stress, anti-cold, anti-allergy, tranquilizing, cardiovascular disease preventing, hypertension preventing, osteopenia preventing, pigmentation preventing and other biological activities. It is also a precursor for the synthesis of the cardiovascular drugs betaxolol and metol. Tyrosol can also be used as a precursor for the synthesis of hydroxytyrosol box salidroside. The hydroxytyrosol has effects of preventing and treating salidroside. Therefore, tyrosol is of interest to researchers as a bioactive in the fine chemicals and pharmaceutical industries. Tyrosol is originally derived from olive oil, but has a low concentration (25mg/kg) in olive oil, and thus the extraction is difficult to be industrially carried out. Chemical synthesis is currently the main production method. But the chemical synthesis process is complicated, the environmental pollution is serious, and the biosynthesis is the most effective alternative method.
Hydroxytyrosol, 3, 4-dihydroxyphenylethanol, is a hydroxylation product of tyrosol. Has stronger antioxidant activity than tyrosol. Because of its high antioxidant activity, it is beneficial to human health, and has activities of cancer chemoprevention, anti-atherosclerosis, inhibiting DNA oxidative damage, protecting skin photodamage, anti-inflammatory and antimicrobial.
Salidroside is the glycosylation product of tyrosol, the main active substance of radix Rhodiolae. The salidroside not only has obvious functions of resisting anoxia, cold, fatigue, microwave radiation, virus and tumor, but also has the effects of enhancing attention, improving working efficiency, delaying organism aging, preventing senile diseases and the like, particularly has very important application value in the aspects of military medicine, aerospace medicine, sports medicine, health care medicine and the like, is an environment-adaptive medicine with development prospect, and is concerned in recent years.
Various invention patents construct recombinant escherichia coli (ZL201310133238.7, ZL20140115011.4, ZL201710091999.9, ZL201711054680.5, ZL201811234765.6, CN202110292484.1 and ZL201910754497.9) and recombinant saccharomyces cerevisiae (ZL201810601213.8, ZL201711479443.3 and cn201910911304.6. the recombinant microorganisms described above achieve de novo synthesis of tyrosol by expression of a keto acid decarboxylase (or an aromatic aldehyde synthase) and an alcohol dehydrogenase.
The production level of the recombinant microorganism is difficult to reach the industrialization level, and needs to be improved.
Disclosure of Invention
The invention aims to provide a new strain for efficiently synthesizing tyrosol, hydroxytyrosol or salidroside, and provides a new method for producing tyrosol, hydroxytyrosol or salidroside.
The technical scheme adopted by the invention is as follows:
in a first aspect of the present invention, there is provided a gene expression module, wherein the gene is: at least one of a tyrosol synthesis pathway enzyme fusion gene, a 4-hydroxyphenylacetate 3-hydroxylase gene, and a glycosyltransferase gene, all of which are Spy/Snoop cyclized.
In some embodiments of the invention, the tyrosol synthesis pathway enzyme fusion gene comprises: aromatic aldehyde synthase AAS (Uniprot Q06086) or α -ketoacid decarboxylase gene, and alcohol dehydrogenase gene.
In some embodiments of the invention, the alpha-keto acid decarboxylase gene is ipdC (Uniprot O07043), ARO10(Uniprot Q06408), KDC4(Uniprot C4R7I0), or PDC (Uniprot C5MDS 4). The alcohol dehydrogenase gene is yahK (Uniprot P75691), ADH6(Uniprot Q04894), ahr (Uniprot A0A024L8S1) or ADH (Uniprot A0A7M3GDV 1).
In some embodiments of the invention, the 4-hydroxyphenylacetate 3-hydroxylase gene is EchpaBC (GenBank ACT46003.1, ACT46002.1), hpaB-hpaC hybrid, or sam5(GenBank ABC 88666.1); the hpaB-hpaC hybrid is not limited to species origin.
In some embodiments of the invention, the glycosyltransferase gene is RrUGT33(Uniprot A0A2I6B3R9), AtUGT85A1(Uniprot W8Q3R5), RsUGT73B6(Uniprot Q6QDB6), RsAS (UniprotKB: Q9AR73), GeGT1(Gene MW015078), or VvGT2(Uniprot G9FKG 7).
In some embodiments of the invention, the Spy is SpyTag/SpyCatcher; the Snoop is a Snoop tag/Snoop captcher.
The amino acid sequences of SpyTag, Snooptag, Spycatcher, SnoopCatcher are as follows:
SpyTag：AHIVMVDAYKPTK；
SnoopTag：KLGDIEFIKVNK；
SpyCatcher：GDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNG；
SnoopCatcher：KPLRGAVFSLQKQHPDYPDIYGAIDQNGTYQNVRTGEDGKLTFKNLSDGKYRLFENSEPAGYKPVQNKPIVAFQIVNGEVRDVTSIVPQDIPATYEFTNGKHYITNEPIPPK。
in a second aspect of the invention, there is provided a recombinant vector comprising a gene expression module according to the first aspect of the invention.
In some embodiments of the invention, the starting vector of the recombinant vector is pZBKBP, pZEABP, pZACBP, pZSABP or any e.
In a third aspect of the present invention, there is provided a recombinant bacterium comprising the gene expression module according to the first aspect of the present invention or the recombinant vector according to the second aspect of the present invention.
In some embodiments of the invention, the recombinant bacterium is any one of:
(a) introducing the tyrosol synthesis pathway enzyme fusion gene into a host bacterium to obtain a recombinant bacterium 1;
(b) introducing the 4-hydroxyphenylacetic acid 3-hydroxylase described in the first aspect of the invention into host bacteria to obtain recombinant bacteria 2;
(c) the glycosyltransferase gene of the first aspect of the present invention is introduced into a host bacterium to obtain a recombinant bacterium 3.
In some embodiments of the invention, the host bacterium is escherichia coli, such as K12-derived bacteria (BW25113, MG1665, W3110, DH10B, BW2952, MDS42 and derivatives thereof), BL21(DE3) and derivatives thereof, BREL606, W, DH1, and the like, and can also be tyrosine-producing escherichia coli, preferably tyrosine-producing escherichia coli.
In some embodiments of the invention, the gene expression module of the first aspect of the invention is introduced into the host bacterium via a recombinant vector.
In a fourth aspect of the invention, there is provided a use of the gene expression module of the first aspect of the invention or the recombinant vector of the second aspect of the invention or the recombinant bacterium of the third aspect of the invention in the preparation of tyrosol, hydroxytyrosol or salidroside.
In a fifth aspect of the present invention, there is provided a method for preparing tyrosol, hydroxytyrosol or salidroside, wherein tyrosol, hydroxytyrosol or salidroside is obtained by fermentation using the recombinant bacterium of any one of the third aspect of the present invention.
In some embodiments of the present invention, the recombinant bacterium according to any one of the third aspect of the present invention is fermented in a medium containing glucose and tyrosine to obtain tyrosol, hydroxytyrosol or salidroside.
In some embodiments of the present invention, the tyrosine may be added exogenously or by adding tyrosine producing bacteria.
In some embodiments of the invention, the tyrosine-producing bacterium can be any tyrosine-producing E.coli, such as the tyrosine-producing bacterium E.coli TYR-14B1(Frontiers in Microbiology,12(2021)710405) disclosed previously in this experiment.
In some embodiments of the invention, the fermentation may employ a single-strain fermentation process, a modular co-culture process, and a modular co-catalysis process.
In some preferred embodiments of the present invention, a modular co-catalysis process is preferred.
In some embodiments of the invention, tyrosol synthesis is modularly co-catalyzed with a tyrosine producing bacterium and recombinant bacterium 1; for the synthesis of hydroxytyrosol, tyrosine producing bacteria, recombinant bacteria 1 and recombinant bacteria 2 are adopted for modular co-catalysis; for salidroside synthesis, tyrosine producing bacteria, recombinant bacteria 1 and recombinant bacteria 3 are adopted for module co-catalysis.
In some embodiments of the invention, the weight ratio of tyrosine producing bacteria to recombinant bacteria 1 for tyrosol synthesis is (1-5): (1-5).
In some embodiments of the invention, the weight ratio of tyrosine-producing bacteria, recombinant bacteria 1 and recombinant bacteria 2 for hydroxytyrosol synthesis is (1-5): (1-5): (1-5).
In some embodiments of the present invention, the weight ratio of the tyrosine-producing bacterium, recombinant bacterium 1, and recombinant bacterium 3 for salidroside synthesis is (1-5): (1-5): (1-5).
In some embodiments of the invention, the inoculum size of the bacteria is: the starting OD600 is 20 to 50.
In some embodiments of the invention, the catalysis condition is biocatalysis at the temperature of 30-42 ℃ and the rpm of 150-250 for 20-40 hours.
The invention has the beneficial effects that:
the invention expresses tyrosol synthesis pathway enzyme fusion in host bacteria to produce tyrosol, introduces 4-hydroxyphenylacetic acid 3-hydroxylase HpaB to produce hydroxytyrosol, and introduces glycosyltransferase to produce salidroside; spy (or Snoop) cyclization can also be treated by heat to increase cell permeability, avoiding mass transport limitations of the cells. The long metabolic pathway is divided into a plurality of pathways through the module co-culture, and different host cells can be used for expression, so that the metabolic burden on the host cells is reduced; moreover, different routes can be selected from the most suitable host cells for expression; the adaptability of the module can be controlled by adjusting the proportion of the module bacteria; plug and play, and is easy to assemble into different ways to synthesize different products. And can also avoid the toxic influence of metabolic intermediates and target products.
Detailed Description
The idea of the invention and the resulting technical effects will be clearly and completely described below in connection with the embodiments, so that the objects, features and effects of the invention can be fully understood. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention.
The amino acid sequences of SpyTag, Snooptag, Spycatcher, Snoopcatcher are as follows:
SpyTag：AHIVMVDAYKPTK(SEQ ID NO.1)；
SnoopTag：KLGDIEFIKVNK(SEQ ID NO.2)；
SpyCatcher：GDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNG(SEQ ID NO.3)；
SnoopCatcher：KPLRGAVFSLQKQHPDYPDIYGAIDQNGTYQNVRTGEDGKLTFKNLSDGKYRLFENSEPAGYKPVQNKPIVAFQIVNGEVRDVTSIVPQDIPATYEFTNGKHYITNEPIPPK(SEQ ID NO.4)。
EXAMPLE 1 construction of Modular bacteria
The tyrosine providing modularity bacterium was a tyrosine producing Escherichia coli, and the tyrosine producing bacterium Escherichia coli TYR-14B1 was a tyrosine modularity bacterium (front ties in Microbiology,12(2021)710405) as disclosed previously in this experiment.
AAS (Uniprot Q06086) and yahK gene (Uniprot P75691) (the sequences of which can be found in the Uniprot database), fusion-expressed with (GSG)2, and SpyTag sequence added to the N-terminus and Spycatcher sequence added to the C-terminus, and the entire gene synthesis SpyTag- (GSG) was carried out by the commercial company2-aas-(GSG)2-yahK-(GSG)2SpyCatcher fragment, NheI/KpnI cleavage ligation of the expression vector pZBKBP (Journal of Industrial Microbiology Biotech) constructed before the laboratoryImmunology 42(2015)627-636), obtaining pZBK-ST-AAS-yahK-SC; transformed into chemically competent cells of Escherichia coli BW25113 to obtain tyrosol modular bacterium Escherichia coli BW25113(pZBK-ST-AAS-yahK-SC), i.e., the recombinant bacterium 1.
EchpaBC (GenBank ACT46003.1, ACT46002.1), SpyTag sequence was added to the N-terminal and Spycatcher sequence was added to the C-terminal, and the commercial company was entrusted to complete SpyTag- (GSG)2-hpaBC-(GSG)2A SpyCatcher fragment, NheI/KpnI enzyme-digested and connected to the pZBKBP vector to obtain pZBK-ST-hpaBC-SC; transformed into chemically competent cells of Escherichia coli BW25113 to obtain hydroxytyrosol modular bacterium Escherichia coli BW25113 (pZBK-ST-hpaBC-SC).
AttUGT 85A1(Uniprot W8Q3R5), SpyTag sequence was added to the N-terminus and SpyCatcher sequence was added to the C-terminus, and the entire Gene Synthesis SpyTag- (GSG) was requested from commercial Co2-AtUGT85A1-(GSG)2A SpyCatcher fragment, wherein NheI/KpnI is enzymatically cut and connected to the pZBKBP vector to obtain pZBK-ST-AtUGT85A 1-SC; transformed into chemically competent cells of Escherichia coli BW25113 to obtain salidroside-blocked bacterium Escherichia coli BW25113(pZBK-ST-AtUGT85A 1-SC).
EXAMPLE 2 Modular Co-catalytic Synthesis of tyrosol
Single colonies of modular tyrosinates and modular tyrosol were picked from LB plates (1% tryptone, 0.5% yeast extract, 1% NaCl), inoculated into LB liquid medium, and cultured at 37 ℃ and 200rpm for about 14 hours. The cells were centrifuged at 2800g for 15min at 4 ℃ for freezing and centrifugation, and washed with cold physiological saline to collect the cells. The tyrosol module was resuspended in 0.1M phosphate buffer pH7.0 to an OD600 of 100 to prepare a bacterial suspension. The bacterial suspension is subjected to heat treatment at 55 ℃ for 20 min. The bacterial suspension is frozen and centrifuged again, and the bacterial suspension is washed with cold physiological saline to collect the bacterial cells. Finally, the tyrosine and tyrosol modular bacteria were resuspended at a ratio of 4:1(g/g) into 10mL of 0.1M phosphate buffer pH7.0 containing 20g/L glucose, 10mM magnesium sulfate and 1.5g/L vitamin C to an OD600 of 25, biocatalysis was carried out at 37 ℃ and 200rpm for 24 h. Tyrosol production was analyzed by HPLC (LC-20A, Shimadzu).
Chromatographic conditions are as follows: chromatographic column Inertsil ODS-SP column (5 μm, 4.6X 150mm, GL Sciences Inc.); mobile phase: a is 0.2% trifluoroacetic acid, and B phase is 100% methanol; gradient elution: the methanol concentration is increased from 14 percent to 45 percent within 0 to 20 min; reducing the methanol concentration from 45% to 14% in 20-30 min; the flow rate is 0.5 mL/min; the column temperature is 30 ℃; the detection wavelength is 280 nm.
As a result, 1.3g/L tyrosol was produced.
Example 3 Modular Co-catalyzed Synthesis of Hydroxytyrosol
Selecting single colonies of the tyrosine modularity bacteria, the tyrosol modularity bacteria and the hydroxytyrosol modularity bacteria on the LB plate, inoculating the single colonies into an LB liquid culture medium, and culturing for about 14 hours at 37 ℃ and 200 rpm. The cells were subjected to refrigerated centrifugation, washed with cold physiological saline, and collected. The bacterial suspension was prepared by resuspending tyrosol-modular bacteria and hydroxytyrosol in 0.1M phosphate buffer, pH7.0, to an OD600 of 100. The bacterial suspension is subjected to heat treatment at 55 ℃ for 20 min. The bacterial suspension is frozen and centrifuged again, and the bacterial suspension is washed with cold physiological saline to collect the bacterial cells. Finally, the tyrosine modularity bacteria, the tyrosol modularity bacteria and the hydroxytyrosol are resuspended in 10mL of 0.1M phosphate buffer solution with pH7.0 and containing 20g/L glucose, 10mM magnesium sulfate and 1.5g/L vitamin C according to the proportion of 4:1:1(g/g) until the OD600 is 25, and the biocatalysis is carried out for 24 hours at 37 ℃ and 200 rpm. Hydroxytyrosol yield was analyzed by HPLC (LC-20A, Shimadzu).
Chromatographic conditions are as follows: chromatographic column Inertsil ODS-SP column (5 μm, 4.6X 150mm, GL Sciences Inc.); mobile phase: a is 0.2% trifluoroacetic acid, and B phase is 100% methanol; gradient elution: the methanol concentration is increased from 14 percent to 45 percent within 0 to 20 min; reducing the methanol concentration from 45% to 14% within 20-30 min; the flow rate is 0.5 mL/min; the column temperature is 30 ℃; the detection wavelength is 280 nm.
As a result, 193.2mg/L hydroxytyrosol was produced.
Example 4 Modular Co-catalytic Synthesis of Salidroside
Selecting single colonies of the tyrosine modularity bacteria, the tyrosol modularity bacteria and the salidroside modularity bacteria on an LB plate, inoculating the single colonies into an LB liquid culture medium, and culturing at 37 ℃ and 200rpm for about 14 h. The cells were subjected to refrigerated centrifugation, washed with cold physiological saline, and collected. The tyrosol and rhodioloside modular bacteria were resuspended in 0.1M phosphate buffer pH7.0 to OD600 of 100 to prepare a bacterial suspension. The bacterial suspension is subjected to heat treatment at 55 ℃ for 20 min. The bacterial suspension was again frozen and centrifuged and the cells were collected by washing with cold physiological saline. Finally, the tyrosine modularity bacteria, the tyrosol modularity bacteria and the salidroside modularity bacteria are resuspended in 10mL of 0.1M phosphate buffer solution with pH value of 7.0 containing 20g/L glucose, 10mM magnesium sulfate and 1.5g/L vitamin C according to the proportion of 4:1:1(g/g) until the OD600 is 25, and the biocatalysis is carried out for 24 hours at 37 ℃ and 200 rpm.
The yield of salidroside was analyzed by HPLC (LC-20A, Shimadzu) as in example 2 to yield 146.1mg/L salidroside.
In addition to the manner mentioned in this example, tyrosol, hydroxytyrosol or salidroside can be produced by means of a modular co-cultivation. Namely, different module bacteria are respectively subjected to seed culture and are inoculated into a fermentation medium according to a certain proportion for fermentation, and a single-bacterium fermentation method can also be adopted. The cyclase can also be directly transformed into a tyrosine-producing bacterium for single-bacterium fermentation. The cyclase gene can also be integrated into the chromosome of Escherichia coli to construct a corresponding recombinant Escherichia coli.
In addition to the manner mentioned in this example, the corresponding enzymes can also be expressed using other expression vectors, and can also be expressed integrated into the E.coli chromosome.
The present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, the embodiments of the present invention and features of the embodiments may be combined with each other without conflict.
SEQUENCE LISTING
<110> Zhongshan university
<120> a recombinant Escherichia coli and method for producing tyrosol, hydroxytyrosol or salidroside by using the same
<130>
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 13
<212> PRT
<213> SpyTag
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Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys
1 5 10
<210> 2
<211> 12
<212> PRT
<213> SnoopTag
<400> 2
Lys Leu Gly Asp Ile Glu Phe Ile Lys Val Asn Lys
1 5 10
<210> 3
<211> 84
<212> PRT
<213> SpyCatcher
<400> 3
Gly Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg Asp Glu Asp Gly
1 5 10 15
Lys Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp Ser Ser Gly Lys
20 25 30
Thr Ile Ser Thr Trp Ile Ser Asp Gly Gln Val Lys Asp Phe Tyr Leu
35 40 45
Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala Pro Asp Gly Tyr
50 55 60
Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn Glu Gln Gly Gln Val
65 70 75 80
Thr Val Asn Gly
<210> 4
<211> 112
<212> PRT
<213> SnoopCatcher
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Lys Pro Leu Arg Gly Ala Val Phe Ser Leu Gln Lys Gln His Pro Asp
1 5 10 15
Tyr Pro Asp Ile Tyr Gly Ala Ile Asp Gln Asn Gly Thr Tyr Gln Asn
20 25 30
Val Arg Thr Gly Glu Asp Gly Lys Leu Thr Phe Lys Asn Leu Ser Asp
35 40 45
Gly Lys Tyr Arg Leu Phe Glu Asn Ser Glu Pro Ala Gly Tyr Lys Pro
50 55 60
Val Gln Asn Lys Pro Ile Val Ala Phe Gln Ile Val Asn Gly Glu Val
65 70 75 80
Arg Asp Val Thr Ser Ile Val Pro Gln Asp Ile Pro Ala Thr Tyr Glu
85 90 95
Phe Thr Asn Gly Lys His Tyr Ile Thr Asn Glu Pro Ile Pro Pro Lys
100 105 110

Claims (10)

1. A gene expression module, comprising: at least one of tyrosol synthesis pathway enzyme fusion gene, 4-hydroxyphenylacetic acid 3-hydroxylase gene and glycosyltransferase gene; the tyrosol synthesis pathway enzyme fusion, 4-hydroxyphenylacetate 3-hydroxylase and glycosyltransferase are all Spy/Snoop cyclised.
2. The gene expression module of claim 1, wherein the tyrosol synthesis pathway enzyme fusion gene comprises: an Aromatic Aldehyde Synthase (AAS)/alpha-ketoacid decarboxylase gene and an alcohol dehydrogenase gene; the alpha-keto acid decarboxylase gene is ipdC, ARO10, KDC4 or PDC; the alcohol dehydrogenase gene is yahK, ADH6, ahr or ADH.
3. The gene expression module of claim 1, wherein the 4-hydroxyphenylacetate 3-hydroxylase gene is EchpaBC, hpaB-hpaC hybrid or sam 5.
4. The gene expression module of claim 1, wherein the glycosyltransferase gene is RrUGT33, AtUGT85a1, RsUGT73B6, RsAS, GeGT1, or VvGT 2.
5. A recombinant vector comprising the gene expression module of any one of claims 1 to 4.
6. A recombinant bacterium comprising the gene expression module according to any one of claims 1 to 4 or the recombinant vector according to claim 5.
7. The recombinant bacterium according to claim 6, wherein the recombinant bacterium is any one of:
(a) introducing the tyrosol synthesis pathway enzyme fusion gene of any one of claims 1 to 4 into a recipient bacterium;
(b) introducing the 4-hydroxyphenylacetic acid 3-hydroxylase gene according to any one of claims 1 to 4 into a recipient bacterium;
(c) a glycosyltransferase gene according to any of claims 1 to 4 introduced into a recipient bacterium.
8. Use of the gene expression module of any one of claims 1 to 4, the recombinant vector of claim 5 or the recombinant bacterium of any one of claims 6 to 7 for the preparation of tyrosol, hydroxytyrosol or salidroside.
9. A method for producing tyrosol, hydroxytyrosol or salidroside, which comprises fermenting the recombinant bacterium of any one of claims 7 to 8.
10. The method of claim 9, wherein the fermentation process is a single-strain fermentation process, a modular co-culture process, or a modular co-catalysis process; preferably a modular co-catalytic process.
CN202210466075.3A 2022-04-29 2022-04-29 Recombinant escherichia coli and method for producing tyrosol, hydroxytyrosol or salidroside by using same Active CN114774451B (en)

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