CN114703116B - Pseudomonas aeruginosa for producing 2-hydroxy phenazine and preparation method and application thereof - Google Patents

Abstract

The invention belongs to the technical field of genetic engineering, and relates to pseudomonas aeruginosa for producing 2-hydroxy phenazine, and a preparation method and application thereof. Aiming at the problem of low yield of 2-hydroxy phenazine produced by a wild pseudomonas aeruginosa strain, the phoB gene, the hppA gene and the OmpR gene in the pseudomonas aeruginosa (Pseudomonas chlororaphis) Qlu-1 are knocked out by using a genetic method; wherein, the preservation number of pseudomonas aeruginosa (pseudomonas chlororaphis) Qlu-1 is CCTCC NO: m2020108. The pseudomonas aeruginosa provided by the invention can obviously improve the yield of 2-hydroxy phenazine. The research shows that the yield of 2-hydroxy phenazine can reach 189mg/L when the pseudomonas aeruginosa treated by genetic engineering is 48 hours, which is improved by about 28 times compared with the wild strain.

Description

Pseudomonas aeruginosa for producing 2-hydroxy phenazine and preparation method and application thereof

Technical Field

The invention belongs to the technical field of genetic engineering, and relates to pseudomonas aeruginosa for producing 2-hydroxy phenazine, and a preparation method and application thereof.

Background

The disclosure of this background section is only intended to increase the understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.

2-hydroxy phenazine (2-OH-PHZ) is an agricultural antibiotic, and 2-hydroxy phenazine can be produced by using a pseudomonas biological fermentation method. The inventor shows that the strain can convert phenazine-1-carboxylic acid into 2-hydroxy phenazine by a biological fermentation method through researching Pseudomonas aeruginosa strain-Pseudomonas Qlu-1 separated from vegetable rhizosphere of a Chinese Weifang, but the yield of the wild strain for 48h is only 6.5mg/L, and the yield is lower.

Disclosure of Invention

In order to solve the defects in the prior art, the invention provides pseudomonas aeruginosa for producing 2-hydroxy phenazine, a preparation method and application thereof.

In order to achieve the above purpose, the technical scheme of the invention is as follows:

in one aspect, a Pseudomonas aeruginosa producing 2-hydroxyphenoxazine is obtained by gene knockout of the phoB gene, hppA gene and OmpR gene in Pseudomonas aeruginosa (Pseudomonas chlororaphis) Qlu-1; wherein, the preservation number of the pseudomonas aeruginosa (Pseudomonas chlororaphis) Qlu-1 is CCTCC NO: m2020108; the preservation unit is China center for type culture collection, which is called CCTCC for short; the preservation time is 8 days of 05 months in 2020; the preservation address is: chinese, university of martial arts, martial arts.

On the other hand, the preparation method of the pseudomonas aeruginosa for producing 2-hydroxy phenazine comprises the process of continuously knocking out target genes in pseudomonas aeruginosa (Pseudomonas chlororaphis) Qlu-1 by adopting a knocking-out method; the target genes are the phoB gene, the hppA gene and the ompR gene;

the knocking-out method comprises the following steps:

obtaining a target-UD fragment according to the target gene and the upstream and downstream sequences connected with the target gene;

constructing a knockout plasmid by using the target-UD fragment;

introducing the knockout plasmid into pseudomonas aeruginosa Qlu-1 by parent hybridization;

screening variant strains knocking out target genes.

In a third aspect, the use of a Pseudomonas aeruginosa as described above for the preparation of 2-hydroxyphenoxazine by biological fermentation.

The beneficial effects of the invention are as follows:

according to the invention, the phoB gene, the hppA gene and the OmpR gene in the pseudomonas aeruginosa Qlu-1 are knocked out to obtain a novel genetic engineering strain, and the expression of the phoB gene, the hppA gene and the OmpR gene in the biological fermentation process is inhibited by the genetic engineering strain, so that the yield of 2-hydroxyphenoxazine prepared by biological fermentation is improved, and specific experiments show that the yield of 2-hydroxyphenoxazine can reach 189mg/L when the pseudomonas aeruginosa subjected to genetic engineering treatment is 48 hours, and is improved by about 28 times compared with a wild strain. In addition, the yield of the 2-hydroxyphenoxazine prepared by the biological fermentation of the pseudomonas aeruginosa can be improved only by a gene knockout method, and the adopted method is simpler and easy to realize, thereby being beneficial to industrialized mass production.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention.

FIG. 1 is a diagram showing the construction of the mutant plasmid pK18-hppA-ud of example 1 of the present invention; (A) amplification of the upstream and downstream homology arms of hppA: 1. amplifying hppA upstream homology arms, 2 amplifying hppA downstream homology arms, 3 amplifying DNA Ladder DL5000; (B) amplification of the hppA upstream and downstream homology arm fusion fragment: 1. amplifying the hppA upstream homology arm fusion fragment, 2, hppA gene upstream and downstream arm fusion fragment, 3, DNA Ladder DL5000.

FIG. 2 is a screen of a plate for double resistance to amphipathic hybridization of the hppA gene of example 1 of the present invention.

FIG. 3 is a photograph of a double-exchanged positive monoclonal image of the screening hppA gene according to the photocopying method of example 1 of the present invention.

FIG. 4 is a PCR-based verification of the hppA knockout strain of example 1 of the present invention; and (3) detecting an external primer: 1. the genome of the hppA knockout strain is used as a template amplified fragment, 2, DNA Ladder,3, blank control, 4, and the genome of the wild strain is used as a template amplified fragment; and (3) detecting an inner primer: 1. the genome of the wild strain is used as a template amplified fragment, 2, DNA Ladder,3, hppA knockout strain is used as a template amplified fragment, 4, blank control.

FIG. 5 is a HPLC chart of 2-OH-PHZ of example 1 of the present invention.

FIG. 6 is a graph showing the yield of 2-OH-PHZ at various times in examples 1-3 of the present invention.

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the present invention. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.

In view of the low yield of 2-hydroxyphenoxazine prepared by the existing wild pseudomonas aeruginosa strain through a biological fermentation method, the invention provides pseudomonas aeruginosa for producing 2-hydroxyphenoxazine, and a preparation method and application thereof.

In an exemplary embodiment of the present invention, there is provided a Pseudomonas aeruginosa producing 2-hydroxyphenoxazine obtained by gene knockout of hpPA gene, phoB gene and OmpR gene in Pseudomonas aeruginosa (Pseudomonas chlororaphis) Qlu-1; wherein, the preservation number of the pseudomonas aeruginosa (Pseudomonas chlororaphis) Qlu-1 is CCTCC NO: m2020108; the preservation unit is China center for type culture collection, which is called CCTCC for short; the preservation time is 8 days of 05 months in 2020; the preservation address is: chinese, university of martial arts, martial arts.

In some examples of this embodiment, the sequences of the hppA gene, the phoB gene and the OmpR gene are shown in SEQ ID No.1, SEQ ID No.7, SEQ ID No.13, respectively.

In another embodiment of the invention, a method for preparing pseudomonas aeruginosa for producing 2-hydroxy phenazine is provided, which comprises the steps of continuously knocking out target genes in pseudomonas aeruginosa (Pseudomonas chlororaphis) Qlu-1 by adopting a knocking-out method; the target genes are hppA gene, phoB gene and OmpR gene;

the knocking-out method comprises the following steps:

obtaining a target-UD fragment according to the target gene and the upstream and downstream sequences connected with the target gene;

constructing a knockout plasmid by using the target-UD fragment;

introducing the knockout plasmid into pseudomonas aeruginosa Qlu-1 by parent hybridization;

screening variant strains knocking out target genes.

In some examples of this embodiment, the sequenced Pseudomonas aeruginosa Qlu-1 genomic sequence is searched for the target gene and its sequence and the fusion fragment target-UD is obtained by PCR fishing and ligating the upstream and downstream fragments of the target gene.

In one or more embodiments, the upstream homology arm primer hppAF1 and hppAR1 of the hppA gene are shown as SEQ ID NO.2 and SEQ ID NO.3 respectively, the downstream homology arm primer hppAF2 and hppAR2 of the hppA gene are shown as SEQ ID NO.4 and SEQ ID NO.5 respectively, and the obtained fusion fragment hppA-UD is shown as SEQ ID NO. 6.

In one or more embodiments, the upstream homology arm primers phoBF1 and phoBR1 of the phoB gene are shown in SEQ ID NO.8 and SEQ ID NO.9 respectively, the downstream homology arm primers phoBF2 and phoBR2 of the phoB gene are shown in SEQ ID NO.10 and SEQ ID NO.11 respectively, and the obtained fusion fragment phoB-UD is shown in SEQ ID NO. 12.

In one or more embodiments, the upstream homology arm primers OmpRF1 and OmpRR1 of the OmpR gene are shown in SEQ ID NO.14 and SEQ ID NO.15 respectively, the downstream homology arm primers OmpRF2 and OmpRR2 of the OmpR gene are shown in SEQ ID NO.16 and SEQ ID NO.17 respectively, and the obtained fusion fragment OmpR-UD is shown in SEQ ID NO. 18.

In some examples of this embodiment, the target-UD fragment is ligated to the plasmid by a cleavage ligation technique to construct a knockout plasmid. The plasmid is preferably a pk18 mobasacb plasmid.

In some examples of this embodiment, the knockout plasmid is introduced into E.coli, and then P.viridis Qlu-1 is co-cultured with E.coli into which the knockout plasmid was introduced, such that the knockout plasmid is introduced into P.viridis Qlu-1.

In some examples of this embodiment, single colonies to be screened are screened by PCR to obtain variants that knock out the gene of interest.

Specifically, gene manipulation is described by taking the seamless knockout of hppA gene as an example.

Extraction of Pseudomonas aeruginosa Qlu-1 genome by using kit

Searching the sequenced Pseudomonas aeruginosa Qlu-1 genome sequence for the hppA gene and its sequence and fishing and ligating the hppA gene upstream and downstream fragments, hppA-UD, by PCR

Construction of the hppA knockout plasmid pK18-hppA-UD by ligation of the hppA-UD fragment to plasmid pK18 mobasacb by the cleavage ligation technique

The pK18-hppA-ud was introduced into the E.coli S17-1 (lambda) strain by means of heat shock transformation,

pseudomonas aeruginosa Qlu-1 and Escherichia coli S17-1 (lambda) were co-cultured and then spread on KB (A) ⁺ K ⁺ ) Introducing pK18-hppA-ud plasmid into Pseudomonas by means of double-parent hybridization, picking single colony growing on double-antibody plate,

single colonies were plated on 15% sucrose KB plates containing ampicillin, and the Qlu-1 knockout hppA strain Qlu-1. DELTA.H was screened and validated by PCR.

In a third embodiment of the invention, there is provided an application of the Pseudomonas aeruginosa in preparing 2-hydroxy phenazine by biological fermentation.

Specifically, the above Pseudomonas aeruginosa was added to a dish containing phenazine-1-carboxylic acid for fermentation.

In order to enable those skilled in the art to more clearly understand the technical scheme of the present invention, the technical scheme of the present invention will be described in detail with reference to specific embodiments.

Example 1

1. Pseudomonas aeruginosa Qlu-1 is inoculated into LB (A+) culture medium, extracted and cultured overnight by shaking table 180rpm at 30 ℃, and the genome of Qlu-1 is extracted by using a genome extraction kit and preserved at-20 ℃ for standby.

2. Searching the sequenced Qlu-1 genome data for the hpPA gene and the sequence upstream and downstream of the gene, and using Qlu-1 strain genome as a template, amplifying an upstream fragment hpPA-U and a line downstream fragment hpPA-D of the hpPA gene (see SEQ ID NO. 1) by using hpPA-F1 (see SEQ ID NO. 2)/hpPA-R1 (see SEQ ID NO. 3) and hpPA-F2 (see SEQ ID NO. 4)/hpPA-R2 (see SEQ ID NO. 5) as primers respectively; the hppA-U and the hppA-D are used as templates, and the hppA-F1/hppA-R1 is used as a template to amplify the hppA upstream and downstream fusion fragment hppA-UD (see SEQ ID NO. 6).

3. The fusion fragment hppA-UD is connected with a knockout plasmid pk18 mobasacb by enzyme digestion to construct a recombinant plasmid pk18-hppA-UD, as shown in FIG. 1.

4. The recombinant plasmid pk18-hppA-UD was introduced into E.coli S17-1 (. Lamda.) by transformation with heat shock

5. Escherichia coli S17-1 (. Lamda.) and Pseudomonas Qlu-1 were subjected to an amphipathic hybridization culture, and the recombinant plasmid pk18-hppA-UD was introduced into Pseudomonas Qlu-1.

6. The Qlu-1 hppA knockout strain was screened together by sucrose plate screening, photocopying screening, PCR screening, etc., and the screened Qlu-1 hppA knockout strain was designated Qlu-1 ΔH as shown in FIGS. 2 to 4.

After fermentation, the extraction with ethyl acetate was examined by HPLC, as shown in FIG. 5, and the accumulated 2-hydroxyphenoxazine in the fermentation broth was increased, wherein the yield of 2-OH-PHZ was increased 5.1 times as high as that of the wild type, to 39.9mg/L, as shown in FIG. 6.

Example 2

Further knockdown of the phoB gene was performed on the basis of Qlu-1.DELTA.H obtained in example 1 to obtain Qlu-1.DELTA.H.DELTA.P.

The procedure for the knockdown of the phoB gene was the same as in example 1 and was as follows:

2. amplifying the upstream fragment phoB-U and the downstream fragment phoB-D of the phoB gene (see SEQ ID NO. 7) by using the primers phoB-F1 (see SEQ ID NO. 8)/phoB-R1 (see SEQ ID NO. 9) and phoB-F2 (see SEQ ID NO. 10)/phoB-R2 (see SEQ ID NO. 11), respectively; phoB-U and PhoB-D were used as templates and PhoB-F1/PhoB-R1 as templates to amplify the PhoB upstream and downstream fusion fragment PhoB-UD (see SEQ ID NO. 12).

3. The fusion fragment phoB-UD is connected with knockout plasmid pk18 mobasacb by enzyme digestion to construct recombinant plasmid pk18-phoB-UD.

4. The recombinant plasmid pk18-phoB-UD was introduced into E.coli S17-1 (. Lamda.) by transformation with heat shock

5. The E.coli S17-1 (. Lamda.) was subjected to an amphipathic hybridization culture with Pseudomonas Qlu-1. DELTA.H, and the recombinant plasmid pk18-phoB-UD was introduced into Pseudomonas Qlu-1. DELTA.H.

6. The Qlu-1 hpppAphoB knockout strain is jointly screened by methods such as sucrose plate screening, photocopying screening, PCR screening and the like, and the screened Qlu-1 hpppAphoB knockout strain is named Qlu-1 delta H delta P.

The 2-OH-PHZ yield of the detected strain was increased to 93mg/L as shown in FIG. 6.

Example 3

The ompR gene was further knocked out based on Qlu-1. DELTA. H.DELTA.P obtained in example 2 to obtain Qlu-1. DELTA.H.DELTA.P.DELTA.O. The procedure for the knockdown of ompR gene was the same as in example 1 and was specifically as follows:

2. amplifying an upstream fragment ompR-U and a line downstream fragment ompR-D of an ompR gene (see SEQ ID NO. 13) by using ompR-F1 (see SEQ ID NO. 14)/ompR-R1 (see SEQ ID NO. 15) and ompR-F2 (see SEQ ID NO. 16)/ompR-R2 (see SEQ ID NO. 17) as primers respectively; the ompR-U and ompR-D are used as templates, and ompR-F1/ompR-R1 is used as a template to amplify an ompR upstream and downstream fusion fragment ompR-UD (see SEQ ID NO. 18).

3. The fusion fragment ompR-UD is connected with knockout plasmid pk18 mobasacb by enzyme digestion to construct recombinant plasmid pk18-ompR-UD.

4. The recombinant plasmid pk18-ompR-UD was introduced into E.coli S17-1 (. Lamda.) by heat shock transformation

5. Coli S17-1 (. Lamda.) was subjected to an amphipathic hybridization culture with Pseudomonas Qlu-1. DELTA.H2P, and the recombinant plasmid pk18-ompR-UD was introduced into Pseudomonas Qlu-1. DELTA.H.DELTA.P.

6. The Qlu-1 hpppAphoBompR knockout strain is jointly screened by methods such as sucrose plate screening, photocopying screening, PCR screening and the like, and the Qlu-1 hpppAphoBompR knockout strain is named as Qlu-1 delta H delta P delta O.

After HPLC detection, the yield of 2-OH-PHZ of the strain was increased to 189mg/L, 29 times that of the original strain Qlu-1, as shown in FIG. 6.

The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

SEQUENCE LISTING

<110> Qilu university of industry

<120> Pseudomonas aeruginosa for producing 2-hydroxy phenazine and preparation method and application thereof

<130>

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aacgctcgcc agccgctgac ccgcgacaag ctgatgaacc tggcccgtgg ccgcgagtgg 600

gatgccctgg agcgttccat cgacgtgcag atttcccgcc tgcgccggat gatcgagccg 660

gatccgtcca agccgcgtta tatccagacg gtctggggtg tgggttatgt gttcgtgccg 720

gatggcacca ccaccaagtg a 741

<210> 14

<211> 27

<212> DNA

<213> artificial sequence

<400> 14

tggaattccg tccgccagtt cgccaag 27

<210> 15

<211> 35

<212> DNA

<213> artificial sequence

<400> 15

gctgctcatc agatgctcct tttatctcgg tgcgc 35

<210> 16

<211> 37

<212> DNA

<213> artificial sequence

<400> 16

aggagcatct gatgagcagc ctgatgattt gtaggag 37

<210> 17

<211> 32

<212> DNA

<213> artificial sequence

<400> 17

caactcgaga tctggttgaa ggcgcggtac ac 32

<210> 18

<211> 1488

<212> DNA

<213> artificial sequence

<400> 18

cgtccgccag ttcgccaagg ccggcttcca gggcgatctg gcccttggtg cggcgctttt 60

gtttgtacgg caggtagagg tcttcgaggc gggtcttggt atcggcgagc ttgatgtcgc 120

gggccagctc cggggtcagc ttgccctgct cctcgatgct ggcgaggatg ctgatgcgcc 180

gttcgtcgag ttctcgcagg tagcgcaggc gctcttccag atgacgcagt tgggtgtcat 240

cgaggctgcc ggtcacttcc ttacggtagc gggcgatgaa gggcacggtg gagccttcat 300

ccaatagagc gacggccgct tcgacctgtt gtgggcgtac accgagttcc tcggcaatgc 360

ggctgttgat gctgtccata aaaccacctg acaaattgtg aaagcaggct cgccggcacg 420

gaaaattggg ctcggcgagt ctggttgagc ggcctggcgt gcgccgctgc ctggatcaaa 480

aggctgcctg ttgacccgtg aaatcgaaca attactgcct ggccgggaaa aataaaaagt 540

ggccacgggc gataacgatc agacgttgcc tgacacaaaa ggcggcgcat tataaccagc 600

gttccagtct tcgggggtat cggcccggtg gcggccgacg gcgggttttc tggcggtcga 660

ggaaaaatct gctaacaatg cacacggtgc gtataacggc agctacgcca taatgcgcac 720

cgagataaaa ggagcatctg atgagcagcc tgatgatttg taggagcgag cgcccgggcc 780

ggcaaacgac ccgggcgctc gcattccgca actctgcgag cgcgactcgc tcctgcaaag 840

cgtgtaatgg ctgtttatga aaaccccgct gtggttcccg caaagcttct tctcccgcac 900

cctctggctg gtgctgatcg tggtgctgtt ctccaaggcg ctgaccctgg tttatctgct 960

gatgaacgag gacgtgctgg tggaccgcca gtacagccac ggcgtggccc tgaccctgcg 1020

cgcctattgg gccgccgacg aatccaaccg cgaccagatc gccgaggccg ccgggctgat 1080

caaggtggtc ggcagcggtg tgccggcggg cgagcagcat tggccctata gcgagattta 1140

tcagcggcag atgcaggccg aattgggcgc cgacactgaa gtgcgcctgc gcatgcacgc 1200

cccgccggcg ctctgggtgc gcgcgccgag cctgggtgat ggctggctga aagtgccgct 1260

gtatccgcat ccgttgcggg ggcagaagat ctggagcgtg ctcggctggt tcctcgcgat 1320

cggcctgttg tccacggctt cggcgtggat cttcgtcagc cagctcaacc agcccctcaa 1380

gcgcctggtc tatgccgccc ggcaactggg gcaggggcgc agcgtgcgcc tgccgatcag 1440

cgatacgccg agcgagatga ccgaggtgta ccgcgccttc aaccagat 1488

Claims (10)

1. A Pseudomonas aeruginosa for producing 2-hydroxy phenazine is characterized in that the Pseudomonas aeruginosa is prepared by a genetic methodPseudomonas chlororaphis) Qlu-1hppAGenes (gene),phoBGene and geneOmpRGene knockout; wherein, pseudomonas aeruginosa is preparedPseudomonas chlororaphis) Qlu-1 has a preservation number of CCTCC NO: m2020108;

hppAgenes (gene),phoBGene and geneOmpRThe sequences of the genes are respectively shown as SEQ ID NO.1, SEQ ID NO.7 and SEQ ID NO. 13.

2. A process for preparing 2-hydroxyphenoxazine from Pseudomonas aeruginosa according to claim 1, comprising knocking out Pseudomonas aeruginosaPseudomonas chlororaphis) Qlu-1, a target gene knockout process; the target gene ishppAGenes (gene),phoBGene and geneOmpRA gene;

the knocking-out method comprises the following steps:

obtaining a target-UD fragment according to the target gene and the upstream and downstream sequences of the connecting target gene;

constructing a knockout plasmid by using the target-UD fragment;

introducing the knockout plasmid into pseudomonas aeruginosa Qlu-1 by parent hybridization;

screening variant strains knocking out target genes.

3. The method for producing 2-hydroxyphenoxazine, according to claim 2, wherein the target gene and the sequence upstream and downstream thereof are searched in the sequenced genomic sequence of Pseudomonas aeruginosa Qlu-1, and the target-UD fragment is obtained by PCR-fishing and ligating the upstream and downstream fragments of the target gene.

4. The process for producing 2-hydroxyphenoxazine according to claim 2,it is characterized in that the method comprises the steps of,hppAgene upstream homology arm primerhppA F1 andhppA r1 is shown as SEQ ID NO.2 and SEQ ID NO.3 respectively,hppAgene downstream homology arm primerhppA F2 andhppA r2 is respectively shown as SEQ ID NO.4 and SEQ ID NO.5, and the obtained fusion fragmenthppA-UD is shown as SEQ ID No. 6;

or alternatively, the first and second heat exchangers may be,phoBgene upstream homology arm primerphoB F1 andphoB r1 is shown as SEQ ID NO.8 and SEQ ID NO.9 respectively,phoBgene downstream homology arm primerphoB F2 andphoB r2 is shown as SEQ ID NO.10 and SEQ ID NO.11 respectively, and the obtained fusion fragmentphoB-UD is shown as SEQ ID No. 12;

or alternatively, the first and second heat exchangers may be,OmpRgene upstream homology arm primerOmpR F1 andOmpR r1 is shown as SEQ ID NO.14 and SEQ ID NO.15 respectively,OmpRgene downstream homology arm primerOmpR F2 andOmpR r2 is shown as SEQ ID NO.16 and SEQ ID NO.17 respectively, and the obtained fusion fragmentOmpR-UD is shown as SEQ ID NO. 18.

5. The method for producing 2-hydroxyphenoxazine, according to claim 2, wherein the target-UD fragment is ligated to a plasmid by a cleavage ligation technique to construct a knockout plasmid.

6. The method for producing 2-hydroxyphenoxazine-producing Pseudomonas aeruginosa according to claim 5, characterized in that the plasmid is a pk18 mobasacb plasmid.

7. The method for producing 2-hydroxyphenoxazine-producing Pseudomonas aeruginosa according to claim 2, characterized in that the knockout plasmid is introduced into Escherichia coli, and then Pseudomonas aeruginosa Qlu-1 is co-cultured with the knockout plasmid-introduced Escherichia coli so that the knockout plasmid is introduced into Pseudomonas aeruginosa Qlu-1.

8. The method for producing 2-hydroxyphenoxazine-producing Pseudomonas aeruginosa according to claim 2, characterized in that single colonies to be screened are screened by PCR to obtain variant strains knocking out target genes.

9. Use of pseudomonas aeruginosa according to claim 1 for the preparation of 2-hydroxyphenoxazine by biological fermentation.

10. The use according to claim 9, wherein the fermentation is carried out by adding the above pseudomonas aeruginosa to a petri dish containing phenazine-1-carboxylic acid.

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