CN102864113A - Strain capable of producing succinic acid, method for producing succinic acid and application thereof - Google Patents

Strain capable of producing succinic acid, method for producing succinic acid and application thereof Download PDF

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CN102864113A
CN102864113A CN2012103920355A CN201210392035A CN102864113A CN 102864113 A CN102864113 A CN 102864113A CN 2012103920355 A CN2012103920355 A CN 2012103920355A CN 201210392035 A CN201210392035 A CN 201210392035A CN 102864113 A CN102864113 A CN 102864113A
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succinic acid
ber208
colon bacillus
hpo
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CN102864113B (en
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姜岷
万青
张常青
梁丽亚
陈旭
苟冬梅
刘嵘明
马江锋
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Nanjing Tech University
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Abstract

The invention discloses a strain capable of producing succinic acid, a method for producing the succinic acid and an application thereof. The strain is named as Escherichichia coil BER208 with the collection number of CCTCCNO: M2012351. The strain grows quickly with glucose serving as the single carbon source in a fermentation medium under the anaerobic condition and is capable of highly producing the succinic acid. The optical density (OD) 6000 reaches 6.87 and the yield of the succinic acid reaches 10.5g/L after fermentation for 48 hours, and compared with an original strain, the yield of the succinic acid is improved by more than two times.

Description

The methods and applications of the bacterial strain of succinic acid-producing and production Succinic Acid thereof
Technical field
The present invention relates to microorganism field, in particular to the bacterial strain of a strain succinic acid-producing and the methods and applications of production Succinic Acid thereof.
Background technology
Succinic Acid (having another name called succsinic acid) is a kind of common natural organic acids, extensively is present in human body, animal, plant and the microorganism.Succinic Acid has widely purposes as a kind of important industrial chemicals in industries such as medicine, food and tensio-active agents.In recent years, day by day serious along with the day by day exhausted and environmental problem of fossil resource, adopt biological synthesis process to replace traditional chemical synthesis to prepare Succinic Acid and get most of the attention, USDOE is classified Succinic Acid as one of biorefinery product of following 12 kinds of most worthies.
The biosynthesizing Succinic Acid is to utilize bacterium, the various microorganisms such as fungi, take glucose or other various hydrolyzed solutions as carbon source, through producing succinic acid by microbial fermentation.Utilize microbe fermentation method to transform renewable resources (glucose, wood sugar etc.) because raw material sources are extensive and cheap, pollute little, environmental friendliness, and can absorb fixation of C O during the fermentation 2, can effectively alleviate Greenhouse effect, therefore become the focus of Recent study.
Fermentation strain is one of biosynthetic key point of Succinic Acid, but most of bacterial strain can only produce the Succinic Acid of low concentration.The fermentation and acid ability that improves Succinic Acid has a lot of methods, yet up to now, under pure anaerobic condition, efficiently utilizes glucose but to rarely have report for the bacterial strain of unicity carbon source and high succinic acid-producing.
Summary of the invention
The object of the present invention is to provide the strain can be under pure anaerobic condition, efficiently utilize glucose be the bacterial strain of unicity carbon source and high succinic acid-producing, with and produce the methods and applications of Succinic Acid.
In order to realize technical purpose of the present invention, the invention provides the bacterial strain of a strain succinic acid-producing, its Classification And Nomenclature be colon bacillus ( Escherichia coli) BER208, its deposit number is: CCTCC NO:M 2012351.
The present invention also provides a kind of method of producing Succinic Acid, the above-mentioned colon bacillus of mentioning that comprises the steps: to ferment ( Escherichia coli) BER208 CCTCC NO:M 2012351, obtain Succinic Acid.
Further, described method comprises: the colon bacillus BER208 of solid plate culture medium culturing is inoculated in to cultivate in the seed culture medium obtains seed liquor; Then seed liquor is inoculated in the fermention medium to obtain Succinic Acid.
Preferably, described method is will be through 37 degrees centigrade of solid plate substratum, and the colon bacillus BER208 that anaerobic condition was cultivated 24 hours is inoculated in seed culture medium, carbonating, at 37 degrees centigrade, anaerobism is cultivated and was obtained seed liquor in 12 hours under 200 rev/mins the condition;
Described seed liquor is inoculated in the described fermention medium according to 2% inoculum size, carbonating, and cultivate 48 hours to obtain Succinic Acid 37 degrees centigrade of anaerobism.
Preferably, described carbonating was carbonating 2 minutes, the anaerobic environment when cultivating to preserve.
Preferably, the prescription of described solid plate substratum is: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, agar 15-20g/L, glucose 10-15g/L.
Preferably, the prescription of described seed culture medium is: peptone 10g/L, yeast powder 5g/L, NaCl 5g/L, glucose 10-15g/L.
Preferably, the prescription of described fermention medium is: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, magnesium basic carbonate 24-42g/L, glucose 30-40g/L.
The present invention also provide colon bacillus ( Escherichia coli) application of BER208 in fermentation production of succinic acid.
The bacterial strain of succinic acid-producing of the present invention is compared with the bacterial strain of existing succinic acid-producing, and its beneficial effect is:
Colon bacillus of the present invention ( Escherichia coli) BER208 CCTCC NO:M 2012351 bacterial strains can be in fermention medium, take glucose as the unicity carbon source under pure anaerobic condition Fast Growth, and high succinic acid-producing: its 48 hours cell densities can reach OD in the anaerobism shaking flask 600=6.87, Succinic Acid output reaches 10.5g/L, and existing starting strain colon bacillus ( Escherichia coli) BER108 CCTCC NO:M 2012068 is under identical condition, the speed of growth is slow, and 48 hours cell densities only are OD 600=3.7, Succinic Acid output is 4.8g/L only, with respect to starting strain, fast growth of the present invention and Succinic Acid output increased more than 2 times, so bacterial strain of the present invention has great social effect and economic worth.
Embodiment
Biomaterial colon bacillus of the present invention ( Escherichia coli) preservation date of BER208 is on 09 14th, 2012, depositary institution's full name is Chinese Typical Representative culture collection center, is called for short CCTCC, the address is: China. and Wuhan. Wuhan University, its deposit number is CCTCC NO:M 2012351.
Starting strain colon bacillus of the present invention ( Escherichia coli) depositary institution's full name of BER108 is Chinese Typical Representative culture collection center, be called for short CCTCC, the address is: China. Wuhan. and Wuhan University, its deposit number is CCTCC NO:M 2012068, its open source is that application number is 201210138292.6, open day is on August 22nd, 2012, and publication number is the Chinese patent application of CN102643770A.
Colon bacillus of the present invention ( Escherichia coli) BER208 is the bacterial strain of high succinic acid-producing.Colon bacillus of the present invention ( Escherichia coli) the BER208 bacterium colony is that circular edge is neat, smooth surface, translucent, small embossment.
Colon bacillus of the present invention ( Escherichia coli) BER208 be with the starting strain colon bacillus ( Escherichia coli) BER108(CCTCC NO:M 2012068) after continuous domestication is cultivated, the solid medium of utilization take glucose as the unicity carbon source is dull and stereotyped, under pure anaerobic condition screening obtain can Fast Growth bacterial strain, again through the anaerobism shake flask fermentation screening obtain can high succinic acid-producing bacterial strain be aimed strain, described screening method can comprise the steps:
1) seed culture medium is cultivated: with the starting strain colon bacillus ( Escherichia coli) BER108(CCTCC NO:M 2012068) utilize seed culture medium to cultivate, 37 ℃, 200r/min cultivates the bacterium liquid that 12h obtains logarithmic phase in liquid amount is the serum bottle of 10mL;
2) cultured continuously domestication mutagenesis: with the bacterium liquid of the logarithmic phase described in the step 1) with 10%(v/v) inoculum size be inoculated in the 500mL bactogen that the 300mL fermention medium is housed, 37 ℃ of heating in water bath pass into the CO of filtration sterilization 2Keep anaerobic environment, and add fresh fermention medium with the speed of 1.5mL/h stream in the culture apparatus.Timing sampling detects the density of thalline in the culture apparatus, and cell density reaches OD in reactor 600=2～3, and keep 48h without larger variation, illustrate that the expression thalline grow to stablize under this flow acceleration, at this moment domestication process is finished a circulation.The flow acceleration of fresh fermention medium is doubled, carry out the cultured continuously of next circulation.Until the flow acceleration of fermention medium reaches 12mL/h, the thalli growth performance is kept stable under this condition;
3) solid plate primary dcreening operation: under aseptic technique, from step 2) take out 2～4mL bacterium liquid the bactogen, with sterilized water dilution 1 * 10 4Doubly, get 100 μ L and be coated on the solid plate, 37 ℃ of anaerobism are cultivated 24h, select growth larger, comparatively full bacterium colony;
4) solid plate sieves again: with the bacterial strain repeatedly turning point cultivation on solid plate that screens in the step 3), 37 ℃ of anaerobism are cultivated 12h, select growth larger, comparatively full bacterium colony;
5) anaerobism shake flask fermentation screening: with the inoculation that filters out in step 4) enlarged culturing in the seed culture medium, 37 ℃, 200r/min cultivates 12h, then ferments in fermention medium, and inoculum size is 2%(v/v), 37 ℃, 200r/min cultivates 48h; Investigate in the bacterium colony that filters out in the step 4) and filter out fast growth, produce the high bacterial strain of acid amount.
Wherein, the prescription of described seed culture medium is: peptone 10g/L, yeast powder 5g/L, NaCl 5g/L, glucose 10-20g/L.
Wherein, the prescription of described fermention medium is: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, magnesium basic carbonate 24-42g/L, glucose 30-40g/L.
Wherein, the prescription of solid plate substratum is: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, agar 15-20g/L, glucose 10-15g/L.
According to following examples, can better understand the present invention.Concrete material proportion described in the case study on implementation, processing condition and result thereof only are used for explanation the present invention, and should also can not limit the present invention described in detail in claims.
Embodiment 1
The present embodiment explanation with colon bacillus ( Escherichia coli) BER108(CCTCC M 2012068) method of carrying out the first step cultured continuously domestication.
The method that the intestinal bacteria original strain carries out the domestication of the first step cultured continuously is as follows:
With the colon bacillus in the cryopreservation tube ( Escherichia coli) BER108(CCTCC M 2012068) as starting strain, in liquid amount is 25mL serum bottle in the 10mL seed culture medium, cultivate, 37 ℃, 200r/min cultivates the bacterium liquid that 12h obtains logarithmic phase; With the bacterium liquid of logarithmic phase with 10%(v/v) inoculum size be inoculated in the 500mL bactogen that the 300mL fermention medium is housed, 37 ℃ of heating in water bath pass into the CO of filtration sterilization 2Keep anaerobic environment, and add fresh fermention medium with the speed of 1.5mL/h stream in the culture apparatus.Timing sampling detects the density of thalline in the culture apparatus, and cell density reaches OD in reactor 600=2～3, and keep 48h without larger variation, illustrate that the expression thalline grow to stablize under this flow acceleration, at this moment domestication process is finished a circulation.The flow acceleration of fresh fermention medium is doubled, carry out the cultured continuously of next circulation.Until the flow acceleration of fermention medium reaches 12mL/h, the thalli growth performance is kept stable under this condition, prepares the screening step of embodiment 2.Wherein, the prescription of described seed culture medium is: peptone 10g/L, yeast powder 5g/L, NaCl 5g/L, glucose 10-20g/L.
The prescription of described fermention medium is: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, magnesium basic carbonate 24-42g/L, glucose 30-40g/L.
Embodiment 2
The present embodiment explanation screening obtain good colon bacillus ( Escherichia coli) method of BER208.
The screening step:
1, solid plate primary dcreening operation
Under aseptic technique, from bactogen, take out 2-4mL bacterium liquid, with sterilized water dilution 1 * 10 4Doubly, get 100 μ L and be coated on the solid plate, 37 ℃ of anaerobism are cultivated 24h, pick out and select fast growth, comparatively full bacterium colony.
2, solid plate sieves again
Bacterial strain repeatedly turning point cultivation on flat board with screening has finally obtained bacterial strain BER10806, BER208, and BER10811 and BER10814 have shown stronger growth velocity and growth stability.
3, shake flask fermentation screening
With enlarged culturing in bacterial strain BER108, BER10806, BER208, BER10811 and the BER10814 access seed culture medium, carbonating 2min, 37 ℃, 200r/min cultivates 12h.Then be inoculated in the fermention medium 100mL anaerobism serum bottle liquid amount 30mL, inoculum size 2%(v/v), carbonating 2min, 37 ℃, 200r/min cultivates 48h.
Wherein, employed culture medium prescription is as follows:
Solid plate substratum: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, agar 15-20g/L, glucose 10-15g/L.
Seed culture medium: peptone 10g/L, yeast powder 5g/L, NaCl 5g/L, glucose 10-20g/L.
Fermention medium: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, magnesium basic carbonate 24-42g/L, glucose 30-40g/L.
Detect each bacterial strain cell density and Succinic Acid output as shown in table 1:
The strain excellent that table 1 screening obtains and the growth of starting strain and product acid are relatively
Bacterial strain OD 600 Succinic Acid output (g/L) Succinic Acid transformation efficiency (%)
BER108 3.7 4.8 73
BER10806 9.48 5.1 81
BER208 6.87 10.5 76
BER10811 3.68 8.5 70
BER10814 5.4 7.4 71
Under pure anaerobic condition, the starting strain colon bacillus ( Escherichia coli) the BER108 poor growth, and Succinic Acid output is also very low, but the bacterial strain colon bacillus that after the bactogen domestication, obtains ( Escherichia coli) BER208, the speed of growth in fermention medium is very fast, and the output of Succinic Acid is also very high, has reached 10.5g/L.
Embodiment 3
The present embodiment explanation screening obtain good colon bacillus ( Escherichia coli) mitotic stability of BER208.
On solid plate, with colon bacillus ( Escherichia coli) many turning points of BER208 cultivate, and will obtain the bacterial strain checking of fermenting respectively, experimental result is as shown in table 2:
Table 2 colon bacillus ( Escherichia coli) analysis of BER208 mitotic stability
Figure 2012103920355100002DEST_PATH_IMAGE002
From experimental result as can be known, through 8 continuous passages, colon bacillus ( Escherichia coli) Succinic Acid output and the Succinic Acid transformation efficiency of BER208 be all comparatively stable, has good mitotic stability, can be used as the production bacterial classification of further research and development.
Embodiment 4
The present embodiment explanation colon bacillus ( Escherichia coli)The method of BER208 fermentation production of succinic acid.
Wherein, the described culture medium prescription of the present embodiment:
Solid plate substratum: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, agar 15-20g/L, glucose 10-15g/L.
Seed culture medium: peptone 10g/L, yeast powder 5g/L, NaCl 5g/L, glucose 10-15g/L.
Fermention medium: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, magnesium basic carbonate 24-42g/L, glucose 30-40g/L.
With colon bacillus ( Escherichia coli)BER208 is seeded on the solid plate, cultivates 37 ℃ of culture temperature, incubation time 24h in anaerobic box.The BER208 that flat board is cultivated is inoculated in the seed culture medium, 25mL serum bottle liquid amount 10mL, and carbonating 2min, 37 ℃ of culture temperature, 200r/min, incubation time 12h obtains seed liquor; Seed liquor is inoculated in the fermention medium inoculum size 2%(v/v), 100mL serum bottle liquid amount 30mL, carbonating 2min, behind the fermentation culture 48h, the output that detects Succinic Acid is 10.5g/L, the Succinic Acid transformation efficiency is 76%.

Claims (9)

1. the bacterial strain of a strain succinic acid-producing, its Classification And Nomenclature be colon bacillus ( Escherichia coli) BER208, its deposit number is: CCTCC NO:M 2012351.
2. a method of producing Succinic Acid is characterized in that, described method comprise the steps: to ferment colon bacillus claimed in claim 1 ( Escherichia coli) BER208, obtain Succinic Acid.
3. method according to claim 2 is characterized in that, described method comprises: the colon bacillus BER208 of solid plate culture medium culturing is inoculated in to cultivate in the seed culture medium obtains seed liquor; Then seed liquor is inoculated in the fermention medium to obtain Succinic Acid.
4. according to claim 2 or 3 arbitrary described methods, it is characterized in that, described method is will be through 37 degrees centigrade of solid plate substratum, the colon bacillus BER208 that anaerobic condition was cultivated 24 hours is inoculated in seed culture medium, carbonating, at 37 degrees centigrade, anaerobism is cultivated and was obtained seed liquor in 12 hours under 200 rev/mins the condition;
Described seed liquor is inoculated in the described fermention medium according to 2% inoculum size, carbonating, and cultivate 48 hours to obtain Succinic Acid 37 degrees centigrade of anaerobism.
5. method according to claim 4 is characterized in that, described carbonating was carbonating 2 minutes.
6. method according to claim 3 is characterized in that, the prescription of described solid plate substratum is: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, agar 15-20g/L, glucose 10-15g/L.
7. method according to claim 3 is characterized in that, the prescription of described seed culture medium is: peptone 10g/L, yeast powder 5g/L, NaCl 5g/L, glucose 10-15g/L.
8. method according to claim 3 is characterized in that, the prescription of described fermention medium is: citric acid 3 gL -1, Na 2HPO 412H 2O 4 gL -1, KH 2PO 48 gL -1, (NH 4) 2HPO 48 gL -1, NH 4Cl 0.2 gL -1, (NH 4) 2SO 40.75 gL -1, MgSO 47H 2O 1 gL -1, CaCl 22H 2O 10.0 mgL -1, ZnSO 47H 2O 0.5 mgL -1, CuCl 22H 2O 0.25 mgL -1, MnSO 4H 2O 2.5 mgL -1, CoCl 26H 2O 1.75 mgL -1, H 3BO 30.12 mgL -1, Al 2(SO4) 31.77 mgL -1, Na 2MoO 42H 2O 0.5 mgL -1, ironic citrate 16.1 mgL -1, magnesium basic carbonate 24-42g/L, glucose 30-40g/L.
Colon bacillus claimed in claim 1 ( Escherichia coli) application of BER208 in fermentation production of succinic acid.
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CN104877941A (en) * 2015-05-27 2015-09-02 南京工业大学 Ammonium ion resistant colibacillus for producing succinic acid and application thereof
CN105543294A (en) * 2016-03-07 2016-05-04 南京工业大学 Method for conversion synthesis of malic acid by using low-activity succinic acid production strains
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CN114621897A (en) * 2022-03-29 2022-06-14 山东理工大学 Strain for producing succinic acid, method for producing succinic acid by using strain and application of strain

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CN104877941A (en) * 2015-05-27 2015-09-02 南京工业大学 Ammonium ion resistant colibacillus for producing succinic acid and application thereof
CN104877941B (en) * 2015-05-27 2018-06-08 南京工业大学 The Escherichia coli of one plant of ammonium ion tolerance type succinic acid-producing and its application
CN105543294A (en) * 2016-03-07 2016-05-04 南京工业大学 Method for conversion synthesis of malic acid by using low-activity succinic acid production strains
CN107227286A (en) * 2017-06-13 2017-10-03 南京工业大学 The genetic engineering bacterium of one plant height production butanedioic acid and its construction method and application
CN107227286B (en) * 2017-06-13 2020-11-03 南京工业大学 Genetically engineered bacterium capable of producing succinic acid at high yield, and construction method and application thereof
CN114395518A (en) * 2021-12-07 2022-04-26 南京工业大学 Recombinant escherichia coli and construction method and application thereof
CN114621897A (en) * 2022-03-29 2022-06-14 山东理工大学 Strain for producing succinic acid, method for producing succinic acid by using strain and application of strain
CN114621897B (en) * 2022-03-29 2024-02-27 山东理工大学 Strain for producing succinic acid and method for producing succinic acid by strain and application of strain

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