CN110157756A - A method of yield of erythrocin is improved by the red mould SACE_0303 gene of the more spores of transformation sugar - Google Patents
A method of yield of erythrocin is improved by the red mould SACE_0303 gene of the more spores of transformation sugar Download PDFInfo
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- CN110157756A CN110157756A CN201910178832.5A CN201910178832A CN110157756A CN 110157756 A CN110157756 A CN 110157756A CN 201910178832 A CN201910178832 A CN 201910178832A CN 110157756 A CN110157756 A CN 110157756A
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
- C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
- C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
Abstract
The invention discloses a kind of methods for improving yield of erythrocin by the red mould SACE_0303 gene of the more spores of transformation sugar, utilize the positive regulator SACE_0303 of the erythromycin biosynthesis of screening, make TetR family open gene SACE_0303 gene overexpression in the more red moulds of spore of sugar by genetic engineering approach, obtain the more red mould erythromycin high-yielding engineering bacterial strains of spore of sugar, erythromycin is produced with the strain fermentation, yield of erythrocin is improved for industrial production and technical support is provided.
Description
Technical field
The present invention relates to genetic engineering fields, more particularly to a kind of pass through, and the more red mould SACE_0303 genes of spore of sugar are transformed
The method for improving yield of erythrocin.
Background technique
Actinomyces can generate secondary metabolite abundant, and these secondary metabolites and its derivative are widely used,
Antibiotic, anticarcinogen, immunosuppressor etc..Erythromycin is that the one kind generated by the red mould of the more spores of sugar is made with broad-spectrum antiseptic
Macrolide antibiotics.Since nineteen fifty-two discovery, erythromycin and its derivative have clinically obtained widely answering
With having developed to the third generation at present.It can be seen that the yield for improving erythromycin is a problem urgently to be solved instantly.
Past industrial producing strain mainly passes through physically or chemically method of mutagenesis acquisition.Traditional induced-mutation technique not only consumes
When, and randomness is larger, can not provide breeding theoretical property guidance.And by the method for genetic engineering, pass through mistake in bacterial strain
Certain controlling genes living or the copy number for increasing certain genes obtain the antibiotic superior strain of transformation, there is good prospect.This
Goal of the invention is exactly that erythromycin superior strain is obtained by genetic engineering approach gene targeted alteration.
In the more red moulds of spore of sugar, controlling gene is not present in erythromycin biological synthesis gene cluster, and passes through genetic engineering
Approach goes the gene of the more red moulds of spore of transformation sugar to obtain superior strain can only be transformed other controlling genes from whole gene group.
2005, prokaryotes transcription regulator was divided by Ramos etc. using sequence similarity, structure and function as standard
LysR、AraC/XylS、TetR、LuxR、LacI、ArsR、IcIR、MerR、AsnC、MarR、NtrC(EBP)、OmpR、DeoR、
Cold shock, GntR and Crp totally 16 families.Wherein TetR family regulatory factor (TetR family
Transcriptional regulators, TFRs) there are the conservative of height, wide participation regulation on DNA binding structural domain
The biological activities such as multi-drug resistance, antibiotic synthesis, osmotic stress reaction.Report in recent years a variety of participation antibiotic yields and
TFRs of streptomycete Morphological Differentiation, such as SAV_576, SAV_3619, SCO1712, AtrA, SLCG_2919 etc., imply TetR
Importance of family's controlling gene in secondary metabolism of Streptomyces and antibiotic biosynthesis.In the more red moulds of spore of sugar, exist
101 TFR, the TFR reported at present for participating in the more red mycotic spore Morphological Differentiations of spore of sugar have SACE_0012 and SACE_7040,
The TFR for participating in regulation erythromycin biosynthesis has SACE_5599, SACE_3986, SACE_7301, SACE_3446, BldD
(SACE_2077), PccD (SACE_3396), SACE_Lrp (SACE_5388), SACE_5754 etc..Show TFRs in erythromycin
Importance in biosynthesis.
Summary of the invention
The object of the invention is to remedy the disadvantages of known techniques, provides a kind of by the more red moulds of spore of transformation sugar
The method of SACE_0303 gene raising yield of erythrocin.
The present invention is achieved by the following technical solutions:
One kind improving yield of erythrocin method by the red mould SACE_0303 gene of the more spores of transformation sugar, passes through genetic engineering
Approach is overexpressed SACE_0303 gene in the more red moulds of spore of sugar, obtains the more red mould erythromycin high-yielding engineering bacterial strains of spore of sugar, uses
The strain fermentation produces erythromycin.
The SACE_0303 gene product can be with positive regulation erythromycin biosynthesis.
The present invention also provides a kind of application of SACE_0303 gene in the more red mould industrial strain WB of spore of sugar, in work
It is overexpressed TetR family open gene SACE_0303 gene in industry superior strain WB, obtains superior strain, it is raw to can be used for erythromycin
It produces.
The invention has the advantages that
The positive regulator SACE_0303 of erythromycin biosynthesis has been screened in present invention research, passes through genetic engineering approach
The more red mould SACE_0303 genes of spore of sugar are overexpressed, the more red mould superior strains of spore of sugar can be obtained, are improved for industrial production red
Mycin yield provides technical support.
Detailed description of the invention
Fig. 1 show the location information of SACE_0303 gene and periphery contiguous gene on chromosome.
Fig. 2 show Δ SACE_0303 mutation construction, wherein (A) is Δ SACE_0303 mutation construction schematic diagram；
(B) it is identified for the PCR of Δ SACE_0303 mutant, M:5000bp DNA Marker；1: Δ SACE_0303 bacterial strain.
Fig. 3 show the yield of erythrocin analysis of wild strain A226 and deletion mutation strain Δ SACE_0303.
Fig. 4 show the covering of SACE_0303 gene, the building for being overexpressed bacterial strain and yield of erythrocin analysis, wherein (A) is
Δ SACE_0303/pIB139-0303 is covered, Δ SACE_0303/pIB139 is unloaded, A226/pIB139-0303 is overexpressed,
The PCR of A226/pIB139 zero load bacterial strain identifies that PCR product is apr resistant gene (776bp)；M:5000bp DNA Marker；
+: pIB139 plasmid；Swimming lane 1: Δ SACE_0303/pIB139 covers unloaded control strain；2: Δ SACE_0303/pIB139-
0303 covering bacterial strain；3:A226/pIB139 is overexpressed unloaded control strain；4:A226/pIB139-0303 is overexpressed bacterium；(B) it is
A226、ΔSACE_0303、ΔSACE_0303/pIB139-0303、ΔSACE_0303/pIB139、A226/pIB139-0303
The yield of erythrocin of bacterial strain is analyzed.
Fig. 5 show the influence of SACE_0303 gene pairs biomass and Morphological Differentiation, wherein (A) is prominent for Δ SACE_0303
Mutant and the mycelial biomass estimation of wild strain A226；It (B) is the spore of Δ SACE_0303 mutant strain and wild strain A226
Sub- growing state.
Fig. 6 show gene transcription level analysis, wherein (A) is erythromycin biosynthetic enzyme genes in Δ SACE_0303
EryAI (SACE_0721) and erythromycin resistance gene ermE (SACE_0733) transcriptional level；(B) in Δ SACE_0303 from
Body SACE_0303 transcriptional level.
Fig. 7 show the expression of SACE_0303 protein purification and analyzes with related gene promoter area EMSA, wherein (A) is
The building of SACE_0303 protein expression vector pET28a-0303；It (B) is SACE_0303 protein purification；It (C) is SACE_0303
The EMSA analysis of albumen and eryAI promoter region, ermE promoter region, SACE_0303 promoter region.
Fig. 8 is shown using SACE_0303 genetic modification erythromycin high industrial production bacterial strain WB is overexpressed, wherein (A) is WB/
The PCR of pIB139-0303 bacterial strain is identified；SACE_0303 gene (420bp) is replaced by tsr resistant gene (1360bp)；M:
5000bp DNA Marker；1:WB/pIB139-0303 bacterial strain；It (B) is erythromycin high industrial production bacterial strain WB and WB/pIB139-
Yield of erythrocin is analyzed in 0303.
Specific embodiment
It elaborates below to the embodiment of the present invention, the present embodiment carries out under the premise of the technical scheme of the present invention
Implement, the detailed implementation method and specific operation process are given, but protection scope of the present invention is not limited to following implementation
Example.
In embodiment using to bacterial strain and plasmid be shown in Table 1.Escherichia coli 37 DEG C LB liquid medium or adding
It is cultivated on the solid LB plate of 1.25% agar.The red mould wild strain A226 of the more spores of erythromycin producing strain sugar and its engineered strain
WB is cultivated in 30 DEG C of pancreas peptone soybean broth (TSBY) culture mediums or on the MGM plate containing 2.2% agar.
Bacterial strain and plasmid used in 1 the present embodiment of table
PEG3350 in embodiment, lysozyme, TES, casamino acid, thiostrepton, apramycin are from Sigma company
Purchase.TSB, yeast extract, peptone purchase are in Oxoid company.Glycine, agar powder, sodium chloride and the examination of other biology
Agent is all purchased from Reagent Company.The general operation technology of Escherichia coli and Str. lincolnensis is according to standard operation.The synthesis of primer and
DNA sequencing is completed by Sangon Biotech (Shanghai) Co., Ltd..
The primer sequence synthesized in experiment is shown in Table 2:
2 list of primers of table
(1) building of SACE_0303 deletion mutant body:
PUCTSR plasmid is that the thiostrepton of the insertion 1360bp between BamHI the and SmaI restriction enzyme site of pUC18 is anti-
Property gene (tsr).By the genome of wild strain A226 as template, with primer SACE_0303up-F/R and SACE_
0303down-F/R amplifies homologous recombination upstream and downstream the homologous fragment 0303-up and 0303-down of about 1500bp.
The two segments 0303-up and 0303-down are successively connected to the tsr resistance gene sequences two of pUCTSR respectively
Building plasmid pUCTS-SACE_0303LR is completed in side；Tsr-0303LR large fragment is converted using homology of chromosome recombinant technique
Into the more red mould A226 of spore of sugar, Positive mutants strain is screened by thiostrepton, obtains SACE_0303 gene by tsr resistance
The engineering strain of gene replacement.It is the positive with plasmid pUCTS-SACE_0303LR using c0303-F/R as identification primer
Template, A226 genome are that negative template carries out PCR identification, and positive deletion mutation strain is named as Δ SACE_0303.
(2) SACE_0303 gene replys the building with overexpression bacterial strain:
By wild strain A226 genome as template, complete SACE_0303 is amplified with covering primer c0303-F/R
Genetic fragment.Electrophoresis is carried out to amplified production and is recycled using kit.By the SACE_0303 segment of recycling and plasmid pIB139
NdeI and XbaI double digestion is carried out, the SACE_0303 segment of double digestion is cloned on digested plasmid pIB139.To the Dan Ke of picking
Integrative plasmid pIB139-0303 is screened in grand progress bacterium solution PCR verifying.
Recombinant plasmid pIB139-0303 is transferred to wild strain A226 protoplast, mutant strain Δ SACE_0303 plasm
In body, pIB139 empty plasmid is transferred to A226 protoplast, in mutant strain Δ SACE_0303 protoplast with method.Conversion
Afterwards about for 24 hours, it cultivates after covering culture medium with apramycin aqueous solution to growing transformant.The transformant of picking carries out expansion training
It supports, a pair of of the Apr primer Apr-p1/p2 designed on pIB139 empty plasmid is recycled to carry out bacterium solution PCR evaluation and screening Δ SACE_
0303/pIB139, Δ SACE_0303/pIB139-0303, A226/pIB139, A226/pIB139-0303 bacterial strain.
(3) in high industrial production bacterial strain WB WB/pIB-0303 erythromycin superior strain building:
The pIB139-0303 plasmid of building is transferred in the more red mould industrial strain WB protoplasts of spore of sugar, WB/ is constructed
PIB-0303 bacterial strain, building screening technique refer to above-mentioned steps.
(4) the HPLC detection of the more red mold fermentation products of spore of sugar:
The more red mould series bacterial strains of spore of inoculation sugar after shaken cultivation 48 hours, are forwarded to R5 into TSB culture medium, at 30 DEG C
Fluid nutrient medium, 30 DEG C shaken cultivation 6 days, then fermentation liquid is extracted using organic solvent, after being evaporated using water-bath,
1mL methanol is added and dissolves and use 0.22 μm of organic filter membrane processing, the Erythromycin A content in rear upper machine examination sample.
(5) the red fungal hyphae body biomass detection of the more spores of sugar:
Δ SACE_0303 mutant strain and A226 are inoculated in the liquid TSR of 50mL, 30 DEG C after shaking table culture 48 hours,
30 DEG C of revolving speed 240rpm shaking table culture 6d in R5 culture medium are transferred to, during which different time sections are set sample, it is clear with dehydrated alcohol
Drying weighs dry cell weight after washing, and repeats sampling twice every time, and obtain average value, is drawn after measurement according to experimental data
Fungal biodiversity curve.
(6) the more red mycotic spore morphologic observations of spore of sugar:
It is marked on R3 culture medium, with identical inoculum concentration respectively by Δ SACE_0303, Δ SACE_0303/
The spore glycerol bacterium solution of pIB139-0303, Δ SACE_0303/pIB139 and wild strain A226, are coated on R3M culture medium
On, it dries to be placed in 30 DEG C of constant incubators in super-clean bench and is inverted growth.Every observing and recording spore growth situation for 24 hours.
(7) in Δ SACE_0303 related gene transcription analysis:
Δ SACE_0303 and the wild strain A226 bacterium solution cultivated for 24 hours are collected, is obtained using full formula gold RNA extracts kit
RNA needed for obtaining, after being inverted to cDNA, uses machine testing on real-time fluorescence quantitative PCR instrument.
(8) protein expression of SACE_0303 and with related gene promoter area EMSA analyze:
According to annotation information on NCBI, design has the protein expression primer p28a- of III restriction enzyme site of NdeI and Hind
0303-F/R is amplified on SACE_0303 complete genome segment connection pET28a.Bacterium solution PCR is carried out to the monoclonal of picking to test
Card.
It is thin that the successful protein expression vector pET28a-0303 of above-mentioned building is transferred to e. coli bl21 (DE3) competence
In born of the same parents, the picking single colonie on Kana resistance LB solid medium expands and carries out bacterium with pET28a-0303-F/R primer after cultivating
Liquid PCR identification, the Strain Designation screened are BL21/pET28a-0303.
BL21/pET28a-0303 is inoculated in Kana resistance LB liquid medium, 37 DEG C are incubated overnight；Next day presses 2%
Inoculum concentration switching, 37 DEG C of cultures to thallus OD600Add IPTG (final concentration 0.5mM) between 0.5-1, is induced under 30 DEG C of 180rpm
Express 5h；Thallus and ultrasonication are collected, centrifuging and taking supernatant carries out Ni- affinity chromatography, passes through after being eluted with the imidazoles of various concentration
SDS-PAGE detection.The albumen size of prediction is 20.57kDa, in addition the sequence about 22kDa on carrier.
Distinguished with eryAI-eryBIV-F/R primer, ermE-eryCI-F/R, SACE_0303-SACE_0304-F/R primer
The promoter region of eryAI, ermE, SACE_0303 gene in PCR amplification erythromycin cluster carries out electrophoresis to amplified production and returns
It receives.Amplified fragments are named as probe PeryAI-eryBIV、PermE-eryCI、PSACE_0303-SACE_0304.By probe respectively with the SACE_ of purifying
0303 albumen is incubated in 30 DEG C, and incubation system carries out activity PAGE detection after being 20 μ L, 10min.
Interpretation of result:
SACE_0303 with close on position of the gene on the more red mould chromosomes of spore of sugar referring to Fig. 1.SACE_0303 gene
Deletion mutation strain A226/ Δ SACE_0303 building process is shown in Fig. 2A.SACE_0303 deletion mutant body is containing 30 μ g/ml
Screening on the R3M plate of thiostrepton and passing through PCR confirms (Fig. 2 B).
A226/ Δ SACE_0303 is fermented in R5 fermentation medium after 6d, detects yield of erythrocin through HPLC, discovery
A226/ Δ SACE_0303 reduces 40% (Fig. 3) compared with the yield of wild strain A226, and HPLC is the result shows that SACE_0303 is ginseng
With positive regulation of erythromycin biosynthesis.
It is due to SACE_0303 gene to further verify the raising of yield of erythrocin in mutant Δ SACE_0303
Caused by missing, by SACE_0303 expression vector pIB139-0303 and pIB139 carrier (control), it is directed respectively into Δ
In the protoplast of SACE_0303 mutant strain and wild strain A226, obtains and reply bacterial strain Δ SACE_0303/pIB139-0303
And unloaded Δ SACE_0303/pIB139, it is overexpressed strains A 226/pIB139-0303 and zero load A226/pIB139.By A226 and
Δ SACE_0303 series bacterial strain carries out shake flask fermentation, and HPLC testing result shows: in Δ SACE_0303 Erythromycin A yield compared with
A226 decline 37%；The yield for replying Erythromycin A in bacterial strain Δ SACE_0303/pIB139-0303 is restored to erythromycin in A226
The level of A；Erythromycin A yield improves 30% (Fig. 4) compared with A226 in A226/pIB139-0303.These are the result shows that SACE_
The biosynthesis of 0303 positive regulation erythromycin.
The Δ SACE_0303 mutant strain of measurement fermentation 6 days and the dry cell weight of A226 bacterial strain, draw corresponding change curve,
Biomass difference of the Δ SACE_0303 compared with A226 less (Fig. 5 A), implies the missing of SACE_0303 gene not as the result is shown
Influence the growth of thallus.
In order to determine whether SACE_0303 gene regulates and controls the sporogenesis of thallus, by mutant strain Δ SACE_0303, covering
Bacterial strain Δ SACE_0303/pIB139-0303, zero load control strain Δ SACE_0303/pIB139 and wild-type strain A226
It is applied on R3M plate simultaneously, 30 DEG C are cultivated 72 hours, and bacterial strain spore growth situation is observed.As the result is shown compared to A226, Δ
The spore shape no significant difference (Fig. 5 B) of SACE_0303 mutant, illustrating SACE_0303 gene delection not influences the shape of spore
At.
QRT-PCR data are shown, compared with wild strain A226, in Δ SACE_0303 mutant strain in erythromycin gene cluster
EryAI, ermE gene transcription level have lowered 3.2 times respectively, 3.3 times (Fig. 6 A), illustrate that SACE_0303 regulation cluster is intragentic
Transcriptional level is to control the biosynthesis of erythromycin；SACE_0303 transcriptional level lowers (Fig. 6 B), shows SACE_0303 egg
The white transcriptional expression that can promote autogene.
The building of SACE_0303 protein expression vector is shown in that Fig. 7 A, protein purification are shown in Fig. 7 B, with eryAI, ermE, SACE_0303
The Binding experiment of promoter region is shown in Fig. 7 C；EMSA analysis shows that SACE_0303 albumen is not combined with eryAI, ermE promoter region,
It is specifically bound with SACE_0303 promoter region, illustrates genetic transcription in SACE_0303 indirect adjustments and controls erythromycin biosynthesis cluster
Expression；It directly facilitates autogene transcription and then regulates and controls erythromycin biosynthesis.
Fig. 8 A is shown in the WB/pIB139-0303 bacterial strain identification of building.By the WB/pIB139-0303 bacterial strain of building and WB bacterial strain
It carries out shaking flask to ferment in parallel, HPLC detects Erythromycin A yield.As the result is shown: red in WB/pIB139-0303 bacterial strain compared to WB
27% (Fig. 8 B) of the output increased of mycin A.
Sequence table
<110>University of Anhui
<120>a kind of method that yield of erythrocin is improved by the red mould SACE_0303 gene of the more spores of transformation sugar
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 564
<212> DNA
<213>the more red moulds of spore (Streptomyces erythraea) of sugar
<400> 1
atgacccgca ccgacgaaag ccgccgtcgc cgagaactgc tgtccaagct gctggagtac 60
tcggccgacc acggcctctc cgaggtgtcg ctgcgcccgc tggccgcagc cgtcgggtcg 120
agcccgcgca tgctgctgta cttcttcggc tccaaggagg gcctcgtccg cgaggtgcac 180
cggcacgccc gcgaggccca gctcgccctc ctggccgacg cgctcggcga caggctcgcg 240
accggctccc gcgccgacgc gctccgcgcg ctgtgggagt ggctgagcga tcccgcgcac 300
cacaacgtcg tgcggttctt cttcgagagc tacgcccgct ccctccacgc acgcgaggac 360
gcctgggagg gcttcggcga ggccagcgtc cgggaatggc tgccgcacat ccggcgggcc 420
ctcgggtcgt cggacgcgga ggcgacgctc gtgctggccg cgctacgcgg cctgatgctc 480
gacctgctcg ccaccggcga caccgaacgg gtgcgcgcgg gcttcgagtc actggtcgcc 540
ggactgccgg tcgagccggc gtag 564
Claims (3)
1. a kind of method for improving yield of erythrocin by the red mould SACE_0303 gene of the more spores of transformation sugar, which is characterized in that logical
Crossing genetic engineering approach makes TetR family open gene SACE_0303 gene overexpression in the more red moulds of spore of sugar, obtains the more spores of sugar
Red mould erythromycin high-yielding engineering bacterial strain produces erythromycin with the strain fermentation.
2. the method according to claim 1, wherein the SACE_0303 gene product can be with positive regulation
Erythromycin biosynthesis.
3. the method according to claim 1, wherein the SACE_0303 gene is in the red mould industry of the more spores of sugar
Application in bacterial strain WB is overexpressed TetR family open gene SACE_0303 gene in high industrial production bacterial strain WB, obtains high yield
Bacterial strain can be used for erythromycin production.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111139192A (en) * | 2019-12-16 | 2020-05-12 | 安徽大学 | Method for improving erythromycin yield by modifying saccharopolyspora erythraea SACE _4682 gene |
| CN111197019A (en) * | 2020-01-10 | 2020-05-26 | 安徽大学 | Method for improving yield of erythromycin through saccharopolyspora erythraea SACE _1906 gene approach |
| CN112458136A (en) * | 2020-12-11 | 2021-03-09 | 安徽大学 | Saccharopolyspora erythraea with modified SACE-1409 gene and application thereof |
| CN114457101A (en) * | 2022-01-12 | 2022-05-10 | 安徽大学 | Method for improving yield of erythromycin by modifying saccharopolyspora erythraea SACE _1558 gene and application of method |
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| CN111139192A (en) * | 2019-12-16 | 2020-05-12 | 安徽大学 | Method for improving erythromycin yield by modifying saccharopolyspora erythraea SACE _4682 gene |
| CN111139192B (en) * | 2019-12-16 | 2022-04-15 | 安徽大学 | Method for improving erythromycin yield by modifying saccharopolyspora erythraea SACE _4682 gene |
| CN111197019A (en) * | 2020-01-10 | 2020-05-26 | 安徽大学 | Method for improving yield of erythromycin through saccharopolyspora erythraea SACE _1906 gene approach |
| CN111197019B (en) * | 2020-01-10 | 2022-04-15 | 安徽大学 | Method for improving yield of erythromycin through saccharopolyspora erythraea SACE _1906 gene approach |
| CN112458136A (en) * | 2020-12-11 | 2021-03-09 | 安徽大学 | Saccharopolyspora erythraea with modified SACE-1409 gene and application thereof |
| CN114457101A (en) * | 2022-01-12 | 2022-05-10 | 安徽大学 | Method for improving yield of erythromycin by modifying saccharopolyspora erythraea SACE _1558 gene and application of method |
| CN114457101B (en) * | 2022-01-12 | 2024-01-12 | 安徽大学 | Method for improving erythromycin yield by modifying rhodosporidium saccharatum SACE_1558 gene and application |
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