CN107058144A - A kind of restructuring yeast strains for producing itaconic acid and its construction method and application - Google Patents
A kind of restructuring yeast strains for producing itaconic acid and its construction method and application Download PDFInfo
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- CN107058144A CN107058144A CN201710080169.6A CN201710080169A CN107058144A CN 107058144 A CN107058144 A CN 107058144A CN 201710080169 A CN201710080169 A CN 201710080169A CN 107058144 A CN107058144 A CN 107058144A
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Abstract
The invention discloses a kind of restructuring yeast strains for producing itaconic acid and its construction method and application.The recombinant yeast inserts the restructuring yeast strains that aconitate decarboxylase gene is obtained using C.glycerolgenesis as starting strain in C.glycerolgenesis genomes.The recombinant plasmid for building the expression cassette Pgap CadA Tgap of decarboxylase gene containing aconitate zeocin is transformed by genetic engineering, expression aconitate decarboxylation enzymatic aconitate in C.glycerolgenesis genomes is inserted after being linearized and obtains itaconic acid.The restructuring yeast strains that the present invention is provided can synthesize itaconic acid, and new approaches are provided for itaconic acid production.
Description
Technical field
The present invention relates to a kind of restructuring yeast strains for producing itaconic acid and its construction method and application, belong to genetic engineering neck
Domain.
Background technology
Itaconic acid is a kind of important chemical products, is 12 important high added value chemistry that USDOE's certification is
One of product, mainly for the production of resin, plastics, rubber, synthetic fibers, surfactant etc..Itaconic acid polymer can also be used
Substitute original additive acrylic acid.In addition, studies have found that itaconic acid can be as a kind of anti-in mammalian immune cell
Fungus matter works.
At present, the production bacterial strain of itaconic acid is mainly Aspergillus terreus.The metabolic pathway of itaconic acid is followed by TCA in Aspergillus terreus
What ring was diverted, the aconitate in TCA circulations is in aconitate decarboxylase (cis-aconitic acid
Decarboxylase, CAD) in the presence of change into itaconic acid (Dwiarti, L., Yamane, K., Yamatani, H.,
Kahar,P.,Okabe,M.,2002.Purification and characterization of cis-aconitic acid
decarboxylase from Aspergillus terreus TN484-M1.J.Biosci.Bioeng.94,29–33；
Kanamasa,S.,Dwiarti,L.,Okabe,M.,Park,E.Y.,2008.Cloning and functional
characterization of the cis-aconitic acid decarboxylase(CAD)gene from
Aspergillus terreus.Appl.Microbiol.Biotechnol.80,223-229), aconitate decarboxylase CAD is
Key enzyme in itaconic acid biosynthesis pathway.However, in itaconic acid building-up process, Aspergillus terreus is slow-growing, fermentation period
It is long；Continuous aeration and high-speed stirred is needed in fermentation process to meet demand of the thalli growth to dissolved oxygen, but high-speed stirred is produced
Mechanical pressure can cause that mycelia is impaired and influences thalli growth again.These shortcomings of Aspergillus terreus urgently to seek a kind of new
Host produces itaconic acid.
Candida glycerolgenesis (C.glycerolgenesis CCTCC AB 204047) is that China possesses independent intellectual production
One plant of power has the industrial strain of excellent fermenting property, and fast growth can be in the hypertonic of 55% glucose or 15%Nacl
Normal growth breeding on pressure culture medium, the characteristics of with resistance to hypertonic and high resistance to cold and diseases, has compared with strong tolerance to itaconic acid thoroughly.
The content of the invention
Present invention aims at provide a kind of restructuring yeast strains for producing itaconic acid and its construction method and application.
To achieve these goals, the technical solution adopted by the present invention is：
A kind of restructuring yeast strains for producing itaconic acid, the bacterial strain is with C.glycerolgenesis CCTCC AB204047
For starting strain, aconitate decarboxylation expression of enzymes is inserted in C.glycerolgenesis genomes by genetic engineering transformation
The restructuring yeast strains that box Pgap-CadA-Tgap-zeocin is obtained.
Heretofore described aconitate decarboxylase expression cassette Pgap-CadA-Tgap-zeocin includes can be thin in host
The DNA of the aconitate decarboxylase shown in SEQ ID NO.1 is expressed in born of the same parents, starts the aconitate decarboxylase gene transcription
C.glycerolgenesis glyceraldehyde-3-phosphate dehydrogenase gene promoters, terminate the transcription of aconitate decarboxylase gene
C.glycerolgenesis glyceraldehyde-3-phosphate dehydrogenase gene terminators, include the DNA, the DNA of expression bleomycin
Promoter and terminator including starting and terminating the bleomycin genetic transcription.
The construction method of the restructuring yeast strains of itaconic acid is produced, the expression of enzymes of decarboxylation containing aconitate is built by genetic engineering
Box Pgap-CadA-Tgap-zeocin recombinant plasmid, C.glycerolgenesis bases are inserted into after recombinant plasmid linearisation
Because obtaining restructuring yeast strains in group.
Application of the restructuring yeast strains of itaconic acid in itaconic acid is prepared is produced, described restructuring yeast strains are incubated at
In fermentation medium, the fermentation medium is：50g/L glucose, 10g/L dusty yeasts, 20g/L peptones, the μ of zeocin 150
G/mL, surplus is water.
Advantage for present invention：What the present invention was provided makes Candida glycerolgenesis synthesize clothing by genetic engineering transformation
The method of health acid, it is motivated, it is workable.The restructuring yeast strains itaconic acid fermentation yield of offer is generally higher than other ferment
Female recombinant bacterial strain (Blazeck J, Miller J, Pan A, et al.Metabolic engineering of
Saccharomyces cerevisiae for itaconic acid production.[J].Applied
Microbiology and Biotechnology,2014,98(19):8155-64；Blazeck J,Hill A,Jamoussi
M,et al.Metabolic engineering of Yarrowia lipolytica for itaconic acid
production.[J].Metabolic Engineering,2015,32:66-73.), and bacterial strain in itself have excellent fermentation
Performance, and have stronger tolerance to itaconic acid, with very strong application value, bacterium can be produced as potential itaconic acid
Strain.
Brief description of the drawings
Fig. 1 is the relevant metabolic pathway of itaconic acid in the Aspergillus terreus mentioned in background technology.
Fig. 2 is carrier pCadA-T plasmid maps provided in an embodiment of the present invention, and wherein CadA is aconitate decarboxylase base
Cause.
Fig. 3 is carrier pURGAPZ plasmid maps provided in an embodiment of the present invention, and Pgap is Candida glycerolgenesis glycerine
Aldehyde -3- phosphate dehydrogenase gene promoters, Tgap terminates for C.glycerolgenesis glyceraldehyde-3-phosphate dehydrogenase genes
Son, ZEOCIN is made up of Pzeocin, zeocin and Tzeocin.
Fig. 4 is carrier pURGAPZC plasmid maps provided in an embodiment of the present invention.
Fig. 5 synthesizes the conditional curve of itaconic acid for restructuring yeast strain ferments.
Embodiment
The present invention is described in further detail below by embodiment.
The structure of the aconitate decarboxylase gene cadA expression cassettes of embodiment 1
The clone of 1.1 Aspergillus terreus aconitate decarboxylase genes
The bacterial strains of Aspergillus terreus CICC 40205 used in the present invention come from Chinese industrial microbial strains preservation tube hub.Root
According to Aspergillus terreus FGSC A1156 genome databases announce information design primer cadA-F (5 '-
AAGGGATCCATGACCAAACAATCTGCGGAC-3 ') and cadA-R (5 '-CGAGGTACCTTATACCAGTGGCGATTTCA-
3 '), using Aspergillus terreus CICC 40205 cDNA as template amplification aconitate decarboxylase gene, through 1.0% Ago-Gel electricity
Swimming testing goal fragment length is about 1.5kb, be connected after purpose product rubber tapping recovery purifying with pMD19-T, acquisition base containing cadA
Because of the recombinant plasmid pCadA-T of sequence.PCadA-T is sequenced, the DNA fragmentation of sequencing result display clone is suitable for Aspergillus terreus
Aconitate decarboxylase gene.It is 100% with the sequence homology that Aspergillus terreus FGSC A1156 have been announced.
1.2 target gene cadA expression vector establishments
Plasmid pCadA-T and plasmid pURGAPZ (laboratory has been built) is extracted, enzyme is carried out with BamHI and KpnI respectively
Cut, and carry out rubber tapping recovery purifying, wherein pCadA-T digestion products reclaim the cadA gene bands that size is about 1.5kb,
PURGAPZ digestion products reclaim the purpose band that size is about 6.2kb.Fragment will be reclaimed to be attached, obtain and contain Pgap-
The recombinant plasmid pURGAPZC of CadA-Tgap-zeocin expression cassettes.
Example 2 prepares the restructuring yeast strains containing Pgap-CadA-Tgap-zeocin expression cassettes
2.1 restructuring yeast strains containing Pgap-CadA-Tgap-zeocin expression cassettes are obtained
1) recombinant plasmid pURGAPZC, HindIII linearization for enzyme restriction is extracted.
2) the ring Candida glycerolgenesis bacterial strain of picking 1 access seed culture medium (dusty yeast 10g/L, peptone 20g/L, Portugal
The μ g/mL of grape sugar 20g/L, zeocin 150, surplus is water) in, at 30 DEG C, under the conditions of 200r/min, shaken cultivation 18h is obtained
Liquid seed.Obtained liquid seed is accessed in seed culture medium by 1% (v/v) inoculum concentration, liquid amount is 10mL/
100mL, it is 30 DEG C to control fermentation temperature, and rotating speed is 200r/min, and culture is until OD600For 1.1mL bacterium nights are taken, bacterium is collected by centrifugation
Body, abandons supernatant, and cell is resuspended in 1mL sterilized waters, centrifuge washing 2 times；" conversion Buffer " is added in the following order：
240 μ L PEG, 36 μ L 1.0mol/L LiAc, 50 μ L linearized fragments add water and complement to 360 μ L；After fully mixing, 42 DEG C of water
Bathe 1h；Thalline is collected by centrifugation, is resuspended in 1mL liquid YEPD, 30 DEG C of 200r/min concussion and cultivates 2h；Thalline, 1M is collected by centrifugation
Shan Li Chun Cheongju are washed 2 times；Take 100 μ L cell suspending liquids to be coated on zeocin screening flat boards, cultivate 2~3d in 30 DEG C, converted
Son.
The 2.2 restructuring yeast strains genotype checkings containing Pgap-CadA-Tgap-zeocin expression cassettes
Picking transformant is in the flat lining out purifying of YEPD, 30 DEG C of culture 18h from screening flat board.Picking single bacterium colony is inoculated with
In seed culture medium, at 30 DEG C, under the conditions of 200r/min, shaken cultivation 18h collects thalline and extracts genome, with GAP-F
(5 '-ATAGGGCCCCACCACAGCAGCACCAAC-3 ') and ura5-R (5 '-CCGCGGATCTCGAGTGAACACCATTGT
ACCAATG-3 ') it is primer, the Pgap-CadA-Tgap-zeocin expression cassettes entered in performing PCR amplification recombinant yeast.Through
0.8% agarose gel electrophoresis result is shown, about 4.2kb purpose fragment can be detected in the transformant selected, to set out
Strain gene group DNA template is not expanded to DNA fragmentation as negative control.Accordingly, it is capable to prove successfully to incorporate in transformant
Pgap-CadA-Tgap-zeocin expression cassettes, i.e. required restructuring yeast strains.
The restructuring yeast strains of example 3 produce itaconic acid
1) seed culture medium is prepared：Glucose 20g/L, dusty yeast 10g/L, the μ g/mL of peptone 5g/L, zeocin 150,
Surplus is water；Itaconic acid fermentation culture medium：Glucose 50g/L, dusty yeast 10g/L, the μ g/ of peptone 20g/L, zeocin 150
ML, surplus is water.
2) above-mentioned gained restructuring yeast strains are inoculated in seed culture medium, at 30 DEG C, under the conditions of 200r/min, vibration
18h is cultivated, liquid seed is obtained.Obtained liquid seed is accessed in fermentation medium by 4% (v/v) inoculum concentration, liquid is filled
30mL/250mL, it is 30 DEG C to control fermentation temperature, and rotating speed is 200r/min, and the time is 48h, fermentation ends.
The content of itaconic acid is detected using high performance liquid chromatography (HPLC) in zymotic fluid.Instrument：The efficient liquid of Agilent
Chromatography (matches somebody with somebody UV-vis detector, Composition distribution and work station)；Chromatographic column：Bio-RAD Aminex HPX-87H
column 300mm×7.8mm；Mobile phase：5mmol/L sulfuric acid；Flow velocity：0.6mL/min, column temperature：60 DEG C, UV-detector
(210nm), the μ L of sample introduction 10.
The zymotic fluid of gained carries out the detection of itaconic acid content after fermentation ends, is analyzed by HPLC, zymotic fluid underpants health
The concentration of acid is about 200mg/L.
Above said content is only the basic explanation under present inventive concept, and according to appointing that technical scheme is made
What equivalent transformation all should belong to protection scope of the present invention.
SEQ ID NO.1
MTKQSADSNAKSGVTSEICHWASNLATDDIPSDVLERAKYLILDGIACAWVGARVPWSEKYVQATMSFEPPGA
CRVIGYGQKLGPVAAAMTNSAFIQATELDDYHSEAPLHSASIVLPAVFAASEVLAEQGKTISGIDVILAAIVGFESG
PRIGKAIYGSDLLNNGWHCGAVYGAPAGALATGKLLGLTPDSMEDALGIACTQACGLMSAQYGGMVKRVQHGFAARN
GLLGGLLAHGGYEAMKGVLERSYGGFLKMFTKGNGREPPYKEEEVVAGLGSFWHTFTIRIKLYACCGLVHGPVEAIE
NLQGRYPELLNRANLSNIRHVHVQLSTASNSHCGWIPEERPISSIAGQMSVAYILAVQLVDQQCLLSQFSEFDDNLE
RPEVWDLARKVTSSQSEEFDQDGNCLSAGRVRIEFNDGSSITESVEKPLGVKEPMPNERILHKYRTLAGSVTDESRV
KEIEDLVLGLDRLTDISPLLELLNCPVKSPLV
Claims (8)
1. a kind of restructuring yeast strains for producing itaconic acid, it is characterised in that：The recombinant yeast is with C.glycerolgenesis
CCTCC AB 204047 are starting strain, are inserted by genetic engineering transformation in C.glycerolgenesis genomes along crow
The restructuring yeast strains that head pyruvate decarboxylase gene expression cassette Pgap-CadA-Tgap-zeocin is obtained.
2. the restructuring yeast strains according to right 1, it is characterised in that described aconitate decarboxylase expression cassette Pgap-
CadA-Tgap-zeocin is by Aspergillus terreus aconitate decarboxylase gene, C.glycerolgenesis glyceraldehyde-3-phosphate dehydrogenations
Enzyme gene promoter and terminator, bleomycin zeocin genes, promoter and terminator composition.
3. the restructuring yeast strains according to right 1 to 2, it is characterised in that：The Aspergillus terreus aconitate decarboxylase gene
Encode a protein：
A, the protein being made up of the amino acid sequence shown in SEQ ID NO.1.
4. the restructuring yeast strains according to right 1 to 2, it is characterised in that：The C.glycerolgenesis glyceraldehyde-
3- phosphate dehydrogenase gene promoters and terminator are respectively connected to encode the nucleotide sequence two ends of aconitate decarboxylase,
It is respectively started and terminates the transcription of aconitate decarboxylase gene.
5. the restructuring yeast strains according to right 1 to 2, it is characterised in that：The bleomycin zeocin is used as expression cassette
In resistance screening mark, the promoter and terminator of the gene be connected to the two ends of gene order, is respectively started and eventually
The only transcription of gene.
6. the construction method of the restructuring yeast strains of itaconic acid is produced according to claim 1 to 5, it is characterised in that：Pass through base
Because of the recombinant plasmid of the engineered structure expression cassette of decarboxylase containing aconitate, it is inserted into after recombinant plasmid linearisation
The restructuring yeast strains of the expression cassette of decarboxylase containing aconitate are obtained in C.glycerolgenesis genomes.
7. application of the restructuring yeast strains according to claim 1 to 6 in itaconic acid is prepared, it is characterised in that：By institute
The recombinant yeast stated is incubated at fermentation medium；And collect the culture medium to separate itaconic acid.
8. application of the restructuring yeast strains according to claim 7 in itaconic acid is prepared, it is characterised in that：The fermentation
Culture medium is：50g/L glucose, 10g/L dusty yeasts, 20g/L peptones, the μ g/mL of zeocin 150, surplus is water.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110527637A (en) * | 2019-07-18 | 2019-12-03 | 中国科学院青岛生物能源与过程研究所 | A kind of Aspergillus terreus bacterial strain producing aconitic acid and its construction method and application |
| CN111944706A (en) * | 2020-08-06 | 2020-11-17 | 中国科学院青岛生物能源与过程研究所 | Recombinant aspergillus terreus strain for producing itaconic acid and construction method and application thereof |
| CN113462588A (en) * | 2021-05-20 | 2021-10-01 | 北京化工大学 | Construction method of yarrowia lipolytica genetic engineering bacteria for producing citric acid or itaconic acid by using acetic acid |
| CN113999807A (en) * | 2021-11-02 | 2022-02-01 | 南京工业大学 | Construction method of recombinant strain and application of recombinant strain in production of itaconic acid |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009027999A (en) * | 2007-07-30 | 2009-02-12 | National Univ Corp Shizuoka Univ | Dna encoding cis-aconitic acid decarboxylase, method for producing the cis-aconitic acid decarboxylase, and method for producing itaconic acid |
| CN101886045A (en) * | 2009-05-11 | 2010-11-17 | 财团法人工业技术研究院 | Genetically modified microorganisms for producing itaconic acid with high yields |
| CN101993899A (en) * | 2009-08-25 | 2011-03-30 | 财团法人工业技术研究院 | Producing itaconic acid in yeast using glycerol as the substrate |
| CN103834582A (en) * | 2012-11-22 | 2014-06-04 | 中国科学院青岛生物能源与过程研究所 | Itaconic acid fermentation yield improvement bacterial strain, construction method thereof and itaconic acid production method using bacterial strain |
| CN104822832A (en) * | 2012-11-23 | 2015-08-05 | 帝斯曼知识产权资产管理有限公司 | Itaconic acid and itaconate methylester production |
| WO2016069849A1 (en) * | 2014-10-30 | 2016-05-06 | Board Of Regents, The University Of Texas System | Engineered fungi for itaconic acid production |
-
2017
- 2017-02-15 CN CN201710080169.6A patent/CN107058144A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009027999A (en) * | 2007-07-30 | 2009-02-12 | National Univ Corp Shizuoka Univ | Dna encoding cis-aconitic acid decarboxylase, method for producing the cis-aconitic acid decarboxylase, and method for producing itaconic acid |
| CN101886045A (en) * | 2009-05-11 | 2010-11-17 | 财团法人工业技术研究院 | Genetically modified microorganisms for producing itaconic acid with high yields |
| CN101993899A (en) * | 2009-08-25 | 2011-03-30 | 财团法人工业技术研究院 | Producing itaconic acid in yeast using glycerol as the substrate |
| CN103834582A (en) * | 2012-11-22 | 2014-06-04 | 中国科学院青岛生物能源与过程研究所 | Itaconic acid fermentation yield improvement bacterial strain, construction method thereof and itaconic acid production method using bacterial strain |
| CN104822832A (en) * | 2012-11-23 | 2015-08-05 | 帝斯曼知识产权资产管理有限公司 | Itaconic acid and itaconate methylester production |
| WO2016069849A1 (en) * | 2014-10-30 | 2016-05-06 | Board Of Regents, The University Of Texas System | Engineered fungi for itaconic acid production |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110527637A (en) * | 2019-07-18 | 2019-12-03 | 中国科学院青岛生物能源与过程研究所 | A kind of Aspergillus terreus bacterial strain producing aconitic acid and its construction method and application |
| CN110527637B (en) * | 2019-07-18 | 2021-10-15 | 中国科学院青岛生物能源与过程研究所 | Monascus strain for producing aconitic acid and construction method and application thereof |
| CN111944706A (en) * | 2020-08-06 | 2020-11-17 | 中国科学院青岛生物能源与过程研究所 | Recombinant aspergillus terreus strain for producing itaconic acid and construction method and application thereof |
| CN113462588A (en) * | 2021-05-20 | 2021-10-01 | 北京化工大学 | Construction method of yarrowia lipolytica genetic engineering bacteria for producing citric acid or itaconic acid by using acetic acid |
| CN113999807A (en) * | 2021-11-02 | 2022-02-01 | 南京工业大学 | Construction method of recombinant strain and application of recombinant strain in production of itaconic acid |
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Application publication date: 20170818 |