CN104630291A - Method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains - Google Patents

Method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains Download PDF

Info

Publication number
CN104630291A
CN104630291A CN201510110556.0A CN201510110556A CN104630291A CN 104630291 A CN104630291 A CN 104630291A CN 201510110556 A CN201510110556 A CN 201510110556A CN 104630291 A CN104630291 A CN 104630291A
Authority
CN
China
Prior art keywords
fermentation
glucose
fermentation system
cgmcc
succinic acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201510110556.0A
Other languages
Chinese (zh)
Inventor
吕涛
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to CN201510110556.0A priority Critical patent/CN104630291A/en
Publication of CN104630291A publication Critical patent/CN104630291A/en
Pending legal-status Critical Current

Links

Classifications

    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/44—Polycarboxylic acids
    • C12P7/46—Dicarboxylic acids having four or less carbon atoms, e.g. fumaric acid, maleic acid

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains, and belongs to the technical field of bioengineering. Two fermenting systems of a genetically engineered bacteria CCTCC NO: M2012041 for producing succinic acid and actinobacillus CGMCC 1593 screened from wild fungi are used for coupling metabolizing; the process comprises the following three steps: 1, preparing a glucose and pentose mixture according to a traditional method for preparing cellulose hydrolysate; 2, fermenting through the genetically engineered bacteria CCTCC NO: M2012041 fermenting system, and 3, fermenting through the actinobacillus CGMCC 1593 fermenting system. The glucose in the cellulose hydrolysate is utilized by the genetically engineered bacteria CCTCC NO: M2012041 in the fermenting system, and the pentose in the cellulose hyrolysate is utilized by the natural bacteria strain CGMCC 1593 in the fermenting system; two fermenting system are used for coupling metabolizing to improve the utilization rate of the glucose and the pentose in the cellulose hyrolysate, and the advantages of two strains are completely considered.

Description

A kind of use two kinds of bacterial strain lotus roots close the method that fermentation realizes succinic acid high yield
Technical field
Use two kinds of bacterial strain lotus roots conjunction fermentations to realize a method for succinic acid high yield, the present invention relates to use two kinds of bacterial strains, the reducing sugar component in two fermentation system efficiency utilization cellulosic hydrolysates, realizes the method for succinic acid high yield, belongs to technical field of bioengineering.
Background technology
Maize straw and southern paddy stalk, the bagasse etc. of north of China are very excellent cellulose resource.Although in recent years, academia was for cellulosic pre-treatment, and the research of the techniques such as enzymatic saccharification is ripe day by day, at present, the real extensive preparative chemistry material of cellulose hydrolysis sugar that uses also is in the primary stage.
This wherein has a crucial problem, and namely no matter Mierocrystalline cellulose is the pretreatment process adopting which kind of mode, the methods such as the combination of such as acidolysis, alkaline hydrolysis and steam explosion or several method; Enzymolysis is carried out in the combination re-using multiple enzyme, such as cellulase, xylosidase, fructose enzyme, dextranase, polygalacturonase, the multiple enzyme such as hemicellulase and combination thereof, and in cellulosic hydrolyzed solution, always having the five carbon hydrolysis sugars (pentose) of 20 wt% ~ 30wt%, this is that most of microorganism is difficult to utilize, and is particularly the genetic engineering bacterium that educational circles skillfully applies at present.How making genetic engineering bacterium possess one again can utilize the pathways metabolism of pentose to be the problem of academic circles at present primary study.
But, many bacteriums based on Natural strains screening, such as metabolism can produce the amber bacillus CGMCC 1593 of succinic acid pentose just can be utilized easily to carry out the metabolism of succinic acid, and the original strain of CGMCC 1593 extracts exactly from the digestive tube of herbivore.
Genetic engineering bacterium also exists many advantages, the single selective of such as meta-bolites, and glucose utilization rate is far above Natural strains, and fermentation efficiency is far above Natural strains.CN102634474B(application number 201210094742.6) disclose Corynebacterium acctoacidophlum ( corynebacterium acetoacidophilum) YF/ Δ ldhcCTCC NO:M2012041, be that a strain typically can the genetic engineering bacterium of high-yield succinic, leavening property is compared as follows table:
Strain name Glucose acid invert ratio Succinic acid selectivity
CCTCC NO：M2012041 95% 99%
CGMCC 1593 80% 90%
How can improve the utilization ratio of fiber hydrolysis sugar during the fermentation, the high advantage of engineering bacteria fermentation efficiency can be played again.This is an extremely significant problem.
Solve such problem, nothing more than several aspect.One is, improve the selectivity of specific enzyme, such as starch is the main raw material preparing glucose at present, uses amylase can guarantee that the Starch Hydrolysis of more than 97% becomes glucose with saccharifying enzyme; Two are, as mentioned, what in genetically engineered bacterial classification, increase a non-glucose utilizes pathways metabolism.
The content that the present invention addresses, it is the terms of settlement increased outside above-mentioned solution route, close by the fermentation system of genetic engineering bacterium and the fermentation system lotus root of Natural strains, in fermentation system, utilize the glucose in cellulose hydrolysis sugar based on genetic engineering bacterium, and Natural strains utilizes the pentose in cellulose hydrolysis sugar in fermentation system.
Summary of the invention
The object of this invention is to provide a kind of use two kinds of bacterial strain lotus roots and close the method that fermentation realizes succinic acid high yield.
Technical scheme of the present invention: a kind of use two kinds of bacterial strain lotus roots close the method that fermentation realizes succinic acid high yield, the genetic engineering bacterium CCTCC NO:M2012041 of succinic acid-producing and screening is utilized to close metabolism from actinobacillus CGMCC 1,593 two fermentation system lotus roots of wild mushroom, improve the utilization ratio of glucose and pentose in cellulose hydrolysis sugar, take into account two kinds of bacterium advantage separately simultaneously.
One strain Corynebacterium acctoacidophlum ( corynebacterium acetoacidophilum) YF/ Δ ldh, this bacterial strain be built by gene Knockout lactate dehydrogenase gene ( ldh) disappearance bacterium, be preserved in China typical culture collection center on February 29th, 2012, preserving number CCTCC NO:M2012041; At CN 102634474A(application number 201210094742.6, denomination of invention: the method for a strain Corynebacterium acctoacidophlum and succinic acid-producing thereof) in openly to disclose.
Actinobacillus succinogenes ( actinobacillus succinogenes) CGMCC 1593, open in Chinese patent ZL 200610038113.6, denomination of invention: a kind of bacterial classification of producing succinic acid by microbial fermentation and method, publication number CN 1814747A, publication date on August 9th, 2006.
Described use two kinds of bacterial strain lotus roots close the method that fermentation realizes succinic acid high yield, and step is:
(1) preparation of cellulose hydrolysis sugar
Glucose and pentose mixture is prepared according to published traditional method preparing cellulosic hydrolysate.Using bagasse as main cellulosic material in the present invention, its composition control is that Mierocrystalline cellulose and hemicellulose level are about 18%, and content of lignin is about 12%, and moisture content 70%, originates from Guangxi.
After cellulosic hydrolysate preparation, wherein glucose and the pentose ratio in total sugar content is controlled as 70% and 30%.
The preparation of fermentation system
(2) according to the fermentation of following content preparation genetic engineering bacterium CCTCC NO:M 2012041 fermentation system:
LB substratum: peptone 10 g/L, yeast powder 5 g/L, NaCl 10 g/L, prepares with deionized water.
Competence substratum: glycine 30 g/L, Tween-80 1 g/L, prepares with deionized water.
Activation medium (LBHIS): yeast powder 2.5 g/L, peptone 5 g/L, NaCl 5 g/L, brain heart infusion 18.5 g/L, sorbyl alcohol 91 g/L, prepares with deionized water.
Seed culture medium: add 5 g/L glucose in LB substratum.
Yeast culture base: glucose 25 g/L, K 2hPO 41.5 g/L, MgSO 47H 2o 0.6 g/L, FeSO 45 mg/L, VitB1 0.2 mg/L, urea 2.5 g/L, corn steep liquor 10 g/L, vitamin H 0.2 mg/L, prepares with deionized water.
Bioconversion medium: K 2hPO 40.5 g/L, KH 2pO 40.5 g/L, MgSO 47H 2o 0.5 g/L, FeSO 46 mg/L, MnSO 44.2 mg/L, vitamin H 0.2 mg/L, VitB1 0.2 mg/L, prepares with deionized water.Substratum all uses deionized water to prepare.
Seed in seed culture medium 30 DEG C, oxygen consumption cultivates 24 h under 200 r/min, then be connected in yeast culture base with the inoculum size of 5% (V/V), 30 DEG C, oxygen consumption cultivates 16 h under 200 r/min, cultured thalline is centrifugal enrichment under 4000r/min, to be suspended in bioconversion medium 30 DEG C, constant temperature oscillation 5h under 200 r/min with the cell concentration of 20 g/L (dry weight).
After bioconversion medium after constant temperature oscillation 5h is centrifugal under the condition of 4000r/min, enrichment lower floor thalline, and be added in cellulosic hydrolysate with the cell concentration of dry weight 20 g/L.Cellulosic hydrolysate total reducing sugar 100g/L, glucose is about 70g/L, and pentose is about 30g/L.After transforming 40h, succinic acid content 66.5g/L.
(3) according to the fermentation of following content preparation actinobacillus CGMCC 1593 fermentation system
Second seed culture medium: peptone 5g/L, corn steep liquor 20g/L, glucose 20g/L, potassium primary phosphate 1.5g/L, Sodium phosphate dibasic 1.5g/L, yeast extract paste 5g/L, prepares with deionized water.
Fermention medium: peptone 5g/L, corn steep liquor 10g/L, glucose 25g/L, potassium primary phosphate 2.5g/L, Sodium phosphate dibasic 2.5g/L, yeast extract paste 5g/L, sodium acetate 1 ~ 5g/L, prepares with deionized water.
Seed one-level is TSB nutrient solution, and be connected in the second seed culture medium after cultivating 14h, inoculum size 4%, after seed culture reaches 7h, accesses nutrient solution in the fermentor tank of anaerobic fermentation system, inoculum size 10%.The pH value using sodium bicarbonate (final sodium bicarbonate concentration is 4%) solution to maintain fermentation system in fermenting process is 5.5 ~ 6.5, and optimum control is 6.0 ~ 6.2, and temperature is 37 ~ 39 DEG C, until glucose content is reduced to 5g/L in system.
The fermentation liquid measure of CGMCC 1593 fermentation system is 15% ~ 30% of the fermentation system fermentation liquid measure of step (2) genetic engineering bacterium CCTCC NO:M 2012041 bacterial classification, and best proportion is 15% ~ 20%.
CCTCC NO:M 2012041 bacterial classification fermentation system by glucose consumption totally after, fermented liquid is warming up to 60 ~ 70 DEG C, the residence time is 30min, makes the thorough inactivation of CCTCC NO:M2012041 bacterial classification in system.By in the fermented liquid of CGMCC 1593 fermentation system access CCTCC NO:M 2012041 spawn fermentation tank, the pH value maintaining fermentation system is 5.5 ~ 6.5, and optimum control is 6.0 ~ 6.2, and temperature is 37 ~ 39 DEG C, after 24h, and in system, reducing sugar content is reduced to 5g/L.
In fermented liquid, reducing sugar content, succinic acid content all use liquid chromatogram measuring.In total fermentation system, glucose content is <2g/L, pentose content <2g/L, comprehensive glucose acid invert ratio >90%.
Beneficial effect of the present invention: the fermentation system of genetic engineering bacterium CCTCC NO:M 2012041 and the fermentation system lotus root of Natural strains CGMCC 1593 close by the present invention, in fermentation system, utilize the glucose in cellulose hydrolysis sugar based on genetic engineering bacterium CCTCC NO:M 2012041, and Natural strains CGMCC 1593 utilizes the pentose in cellulose hydrolysis sugar in fermentation system.Two fermentation system lotus roots close metabolism, improve the utilization ratio of glucose and pentose in cellulose hydrolysis sugar, have taken into account two kinds of bacterium advantage separately simultaneously.
Embodiment
Embodiment 1
Fermentation system according to following content preparation CCTCC NO:M 2012041 bacterial classification:
LB substratum 1mL: peptone 10 g/L, yeast powder 5 g/L, NaCl 10 g/L, prepares with deionized water.
Competence substratum 1mL: glycine 30 g/L, Tween-80 1 g/L, prepare with deionized water.
Activation medium (LBHIS) 2mL: yeast powder 2.5 g/L, peptone 5 g/L, NaCl 5 g/L, brain heart infusion 18.5 g/L, sorbyl alcohol 91 g/L, prepares with deionized water.
Seed culture medium 5mL: add 5 g/L glucose in LB substratum.
Yeast culture base 100mL: glucose 25 g/L, K 2hPO 41.5 g/L, MgSO 47H 2o 0.6 g/L, FeSO 45 mg/L, VitB1 0.2 mg/L, urea 2.5 g/L, corn steep liquor 10 g/L, vitamin H 0.2 mg/L, prepares with deionized water.
Bioconversion medium 500mL:K 2hPO 40.5 g/L, KH 2pO 40.5 g/L, MgSO 47H 2o 0.5 g/L, FeSO 46 mg/L, MnSO 44.2 mg/L, vitamin H 0.2 mg/L, VitB1 0.2 mg/L, prepares with deionized water.Substratum all uses deionized water to prepare.
Cellulose hydrolysis sugar concentrated solution 500mL: glucose 140g/L, pentose 60g/L.
Seed in seed culture medium 30 DEG C, oxygen consumption cultivates 24 h under 200 r/min, then be connected in yeast culture base with the inoculum size of 5% (V/V), 30 DEG C, oxygen consumption cultivates 16 h under 200 r/min, cultured thalline is centrifugal enrichment under 4000r/min, to be suspended in bioconversion medium 30 DEG C, constant temperature oscillation 5h under 200 r/min with the cell concentration of 20 g/L (dry weight).After bioconversion medium after constant temperature oscillation 5h is centrifugal under the condition of 4000r/min, enrichment lower floor thalline, and be added in cellulosic hydrolysate with the cell concentration of dry weight 20 g/L.
According to following content preparation CGMCC 1593 fermentation system:
Second seed culture medium 15mL: peptone 5g/L, corn steep liquor 20g/L, glucose 20g/L, potassium primary phosphate 1.5g/L, Sodium phosphate dibasic 1.5g/L, yeast extract paste 5g/L, prepares with deionized water.
Fermention medium 150mL: peptone 5g/L, corn steep liquor 10g/L, glucose 25g/L, potassium primary phosphate 2.5g/L, Sodium phosphate dibasic 2.5g/L, yeast extract paste 5g/L, sodium acetate 1 g/L, prepares with deionized water.
Seed one-level is TSB nutrient solution, and be connected in the second seed culture medium after cultivating 14h, inoculum size 4%, after seed culture reaches 7h, accesses nutrient solution in the fermentor tank of anaerobic fermentation system, inoculum size 10%.The pH value using sodium bicarbonate (final sodium bicarbonate concentration is 4%) to maintain fermentation system in fermenting process is 5.5 ~ 6.5, and temperature is 37 ~ 39 DEG C.
The fermentation liquid measure of CGMCC 1593 fermentation system is 15% of the fermentation system of CCTCC NO:M 2012041 bacterial classification.CCTCC NO:M 2012041 bacterial classification fermentation system by glucose consumption totally after, fermented liquid is warming up to 60 ~ 70 DEG C, the residence time is 30min, makes the thorough inactivation of CCTCC NO:M 2012041 bacterial classification in system.By in the fermented liquid of CGMCC 1593 fermentation system access CCTCC NO:M 2012041 spawn fermentation tank, the pH value maintaining fermentation system is 5.5 ~ 6.5, and temperature is 37 ~ 39 DEG C, after 24k, and in system, reducing sugar content is reduced to 5g/L.
In fermented liquid, reducing sugar content, succinic acid content all use liquid chromatogram measuring.Succinic acid content 82.5g/L in total fermentation system, glucose content is <2g/L, pentose content <2g/L, and comprehensive glucose acid invert ratio is about 92%.

Claims (5)

1. one kind uses two kinds of bacterial strain lotus roots conjunction fermentations to realize the method for succinic acid high yield, it is characterized in that utilizing the genetic engineering bacterium CCTCC NO:M2012041 of succinic acid-producing and screening to close metabolism from actinobacillus CGMCC 1,593 two fermentation system lotus roots of wild mushroom, improve the utilization ratio of glucose and pentose in cellulose hydrolysis sugar, take into account two kinds of bacterium advantage separately simultaneously.
2. use according to claim 1 two kinds of bacterial strain lotus roots close the method that fermentation realizes succinic acid high yield, it is characterized in that step is:
(1) preparation of cellulose hydrolysis sugar
Glucose and pentose mixture is prepared according to traditional method preparing cellulosic hydrolysate, using bagasse as cellulosic material in the present invention, its composition control is Mierocrystalline cellulose and hemicellulose level is 18%, and content of lignin is 12%, moisture content 70%, originates from Guangxi;
After cellulosic hydrolysate preparation, wherein glucose and the pentose ratio in total sugar content is controlled as 70% and 30%;
(2) fermentation of genetic engineering bacterium CCTCC NO:M 2012041 fermentation system
LB substratum: peptone 10 g/L, yeast powder 5 g/L, NaCl 10 g/L, prepares with deionized water;
Competence substratum: glycine 30 g/L, Tween-80 1 g/L, prepares with deionized water;
Activation medium LBHIS: yeast powder 2.5 g/L, peptone 5 g/L, NaCl 5 g/L, brain heart infusion 18.5 g/L, sorbyl alcohol 91 g/L, prepares with deionized water;
Seed culture medium: add 5 g/L glucose in LB substratum;
Yeast culture base: glucose 25 g/L, K 2hPO 41.5 g/L, MgSO 47H 2o 0.6 g/L, FeSO 45 mg/L, VitB1 0.2 mg/L, urea 2.5 g/L, corn steep liquor 10 g/L, vitamin H 0.2 mg/L, prepares with deionized water;
Bioconversion medium: K 2hPO 40.5 g/L, KH 2pO 40.5 g/L, MgSO 47H 2o 0.5 g/L, FeSO 46 mg/L, MnSO 44.2 mg/L, vitamin H 0.2 mg/L, VitB1 0.2 mg/L, prepares with deionized water;
Seed in seed culture medium 30 DEG C, oxygen consumption cultivates 24 h under 200 r/min; Then be connected in yeast culture base with the inoculum size of 5% V/V, 30 DEG C, oxygen consumption cultivates 16 h under 200 r/min; Cultured thalline is centrifugal enrichment under 4000r/min, is suspended in bioconversion medium with the cell concentration of dry weight 20 g/L, 30 DEG C, constant temperature oscillation 5h under 200 r/min;
After bioconversion medium after constant temperature oscillation 5h is centrifugal under the condition of 4000r/min, enrichment lower floor thalline, and be added in cellulosic hydrolysate with the cell concentration of dry weight 20 g/L, cellulosic hydrolysate total reducing sugar 100g/L, glucose controls as 70g/L, pentose controls as 30g/L, after transforming 40h, and succinic acid content 66.5g/L;
(3) fermentation of actinobacillus CGMCC 1593 fermentation system
Second seed culture medium: peptone 5g/L, corn steep liquor 20g/L, glucose 20g/L, potassium primary phosphate 1.5g/L, Sodium phosphate dibasic 1.5g/L, yeast extract paste 5g/L, prepares with deionized water;
Fermention medium: peptone 5g/L, corn steep liquor 10g/L, glucose 25g/L, potassium primary phosphate 2.5g/L, Sodium phosphate dibasic 2.5g/L, yeast extract paste 5g/L, sodium acetate 1 ~ 5g/L, prepares with deionized water;
Seed one-level is TSB nutrient solution, be connected in the second seed culture medium after cultivating 14h, inoculum size 4%, after seed culture reaches 7h, accessed by nutrient solution in the fermentor tank of anaerobic fermentation system, inoculum size 10%, the pH value using sodium hydrogen carbonate solution to maintain fermentation system in fermenting process is 5.5 ~ 6.5, temperature is 37 ~ 39 DEG C, until glucose content is reduced to 5g/L in system;
The fermentation liquid measure of CGMCC 1593 fermentation system is 15% ~ 30% of the fermentation system fermentation liquid measure of step (2) genetic engineering bacterium CCTCC NO:M 2012041 bacterial classification;
CCTCC NO:M 2012041 bacterial classification fermentation system by glucose consumption totally after, fermented liquid is warming up to 60 ~ 70 DEG C, the residence time is 30min, makes the thorough inactivation of CCTCC NO:M 2012041 bacterial classification in system; By in the fermented liquid of CGMCC 1593 fermentation system access CCTCC NO:M 2012041 spawn fermentation tank, the pH value maintaining fermentation system is 5.5 ~ 6.5, and temperature is 37 ~ 39 DEG C, after 24h, and in system, reducing sugar content is reduced to 5g/L;
In fermented liquid, reducing sugar content, succinic acid content all use liquid chromatogram measuring, and in total fermentation system, glucose content is <2g/L, pentose content <2g/L, comprehensive glucose acid invert ratio >90%.
3. use according to claim 2 two kinds of bacterial strain lotus roots close the method that fermentation realizes succinic acid high yield, it is characterized in that the fermentation of step (3) actinobacillus CGMCC 1593 fermentation system, the pH value using sodium hydrogen carbonate solution to maintain fermentation system in fermenting process is 6.0 ~ 6.2, temperature is 37 ~ 39 DEG C, until glucose content is reduced to 5g/L in system.
4. use according to claim 2 two kinds of bacterial strain lotus roots close the method that fermentation realizes succinic acid high yield, it is characterized in that the fermentation of step (3) actinobacillus CGMCC 1593 fermentation system, the fermentation liquid measure of CGMCC 1593 fermentation system is 15% ~ 20% of the fermentation system fermentation liquid measure of genetic engineering bacterium CCTCC NO:M 2012041 bacterial classification.
5. use according to claim 2 two kinds of bacterial strain lotus roots close the method that fermentation realizes succinic acid high yield, it is characterized in that the fermentation of step (3) actinobacillus CGMCC 1593 fermentation system, by in the fermented liquid of CGMCC 1593 fermentation system access CCTCC NO:M 2012041 spawn fermentation tank, the pH value maintaining fermentation system controls 6.0 ~ 6.2, temperature is 37 ~ 39 DEG C, after 24h, in system, reducing sugar content is reduced to 5g/L.
CN201510110556.0A 2015-03-13 2015-03-13 Method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains Pending CN104630291A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510110556.0A CN104630291A (en) 2015-03-13 2015-03-13 Method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510110556.0A CN104630291A (en) 2015-03-13 2015-03-13 Method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains

Publications (1)

Publication Number Publication Date
CN104630291A true CN104630291A (en) 2015-05-20

Family

ID=53209535

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510110556.0A Pending CN104630291A (en) 2015-03-13 2015-03-13 Method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains

Country Status (1)

Country Link
CN (1) CN104630291A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107228819A (en) * 2017-06-04 2017-10-03 胥振国 A kind of flow cytometry assays of staphylococcus aureus

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101215582A (en) * 2007-12-28 2008-07-09 江南大学 Method for producing succinic acid by fermenting straw raw material
CN102634474A (en) * 2012-03-31 2012-08-15 江南大学 Corynebacterium acetoacidophilum strain and method for producing succinic acid therefrom

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101215582A (en) * 2007-12-28 2008-07-09 江南大学 Method for producing succinic acid by fermenting straw raw material
CN102634474A (en) * 2012-03-31 2012-08-15 江南大学 Corynebacterium acetoacidophilum strain and method for producing succinic acid therefrom

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
陈洪章，王岚: "《生物质生化转化技术》", 31 October 2012 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107228819A (en) * 2017-06-04 2017-10-03 胥振国 A kind of flow cytometry assays of staphylococcus aureus
CN107228819B (en) * 2017-06-04 2019-12-13 胥振国 Flow cytometry detection method for staphylococcus aureus

Similar Documents

Publication Publication Date Title
Hernández et al. Saccharification of carbohydrates in microalgal biomass by physical, chemical and enzymatic pre-treatments as a previous step for bioethanol production
CN106636226B (en) Method for preparing butanol by fermenting lignocellulose
JP5711873B2 (en) Simultaneous saccharification and fermentation of cellulosic materials
CN101851656B (en) Method for producing cellulosic ethanol
Liu et al. Consolidated bioprocess for bioethanol production with alkali-pretreated sugarcane bagasse
CN101608192B (en) Method for producing succinic acid employing corn cob
CN106086085A (en) Utilize the method that bagasse cellulose and cane molasses produce ethanol
CN106957876B (en) Method for preparing butanol by fermenting lignocellulose raw material
Ra et al. Optimization of pretreatment conditions and use of a two-stage fermentation process for the production of ethanol from seaweed, Saccharina japonica
Shaibani et al. Ethanol production from sugarcane bagasse by means of enzymes produced by solid state fermentation method
CN102864082B (en) Method for culturing citric acid fermenting seeds and method for preparing citric acid by fermenting
CN102703543B (en) Method for preparing bacterial cellulose by tuberous raw materials
CN112646846B (en) Method for producing beta-1,3-glucan by using non-growth coupling characteristic of bacteria
CN103614448B (en) Method for preparing bioethanol by taking sodium alginate or algae as active ingredients
CN107760753A (en) Method for producing butanol by co-culture fermentation of high-temperature anaerobe for pyrolyzing sugar and clostridium acetobutylicum
CN105779328A (en) Method for culturing clostridium thermocellum
CN101703152A (en) Method for preparing astaxanthin feed additive by using bear spent grains
CN103789360A (en) Method for preparing fumaric acid fermentation liquor by fermenting corncob cellulose
CN114807098B (en) Culture method for producing extracellular cellulose degrading enzyme system
CN111118071A (en) Fermentation method for producing xylitol and ethanol by using non-detoxified cellulose raw material
CN104630291A (en) Method for realizing high-yield succinic acid through coupling fermenting of two bacterial strains
JP2014176351A (en) Method for producing ethanol
BR112014031261B1 (en) COMPOSITION OF FERMENTATION BROTH, METHOD FOR CONTROLLING BACTERIAL CONTAMINATION, METHOD FOR PRODUCTION OF ETHANOL AND METHOD FOR IMPROVING ZYMOMONA CELL GROWTH
CN105624213B (en) A method of 2,3- butanediol is produced using microalgae for raw material
CN102851325A (en) Fermentation method for producing ethanol by using enzymatic saccharification of corn cob

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20150520