CN101565721A - Method for producing 2,3-butanediol by using starch raw materials - Google Patents
Method for producing 2,3-butanediol by using starch raw materials Download PDFInfo
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- CN101565721A CN101565721A CNA2009100154009A CN200910015400A CN101565721A CN 101565721 A CN101565721 A CN 101565721A CN A2009100154009 A CNA2009100154009 A CN A2009100154009A CN 200910015400 A CN200910015400 A CN 200910015400A CN 101565721 A CN101565721 A CN 101565721A
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- butyleneglycol
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Abstract
The invention discloses a method for producing 2,3-butanediol by using starch raw materials, which comprises the following steps: subjecting the starch raw materials to treatment by alpha-amylase and maltogenic amylase; and producing the 2,3-butanediol by carrying out separate hydrolysis and fermentation or simultaneous sacchrification and fermentation under an aseptic condition, using saccharification liquid or liquefaction liquid as a fermentable substrate and using Klesiella pneumoniae as a fermentable strain. Finally the concentration of the 2,3-butanediol reaches 43 to 112g/L, and the final concentration of acetoin and the final concentration of the 2,3-butanediol reach 45 to 121g/L. The method has the advantages of using the starch raw materials to produce a high-value platform compound by fermentation, reducing the raw material cost of the production of the 2,3-butanediol by the fermentation and establishing the optimal fermentation condition for the production of the 2,3-butanediol by the sacchrification and fermentation of the starch raw materials. The production of the 2,3-butanediol by the simultaneous sacchrification and fermentation of the invention can save energy and equipment investment, reduce fermentation period and improve production efficiency.
Description
Technical field
The invention belongs to technical field of biochemical industry, relate in particular to a kind of starch materials that utilizes and produce 2, the method for 3-butyleneglycol.
Background technology
2, and the 3-butyleneglycol (2, be a kind of hardware and software platform compound of high added value 3-butanediol), have wide application field.2, the 3-butyleneglycol is a kind of chipal compounds of colorless and odorless, and 3 kinds of configurations are arranged.Boiling point is 180-184 ℃, and zero pour is lower, D-2, and 3-butyleneglycol (60 ℃) can be used as antifreezing agent.2, the 3-butyleneglycol has higher octane value, can be used to produce senior aviation kerosene, and be valuable liquid fuel.Dehydration can be converted into the industrial solvent methylethylketone, and methylethylketone can be used as fuel dope, and can be used as solvent and be used for resin and system lacquer industry.The further dehydration of methylethylketone can obtain 1,3-butadiene, can be used as substrate be used for China Synthetic Rubber Industry (Syu MJ.Appl Microbiol Biotechnol, 2001,55:10-18).2, the 3-butanediol dehydrogenation can obtain acetoin and dimethyl diketone, can be used as spices as foodstuff additive.In China, 2, the 3-butyleneglycol also is added in the liquor to improve liquor flavor.2, the 3-butyleneglycol also have in fields such as printing ink, makeup, fragrance fumette, tenderizer, softening agent, explosive and pharmaceutical carriers potential use (Xiu ZL, Zeng AP.Appl Microbiol Biotechnol, 2008,78:917-926).In recent years because the instability of oil price and rising steadily of petroleum chemicals price, 2, the price of 3-butyleneglycol and produce the concern that also causes in the world more and more widely.Develop cheap renewable biomass resource fermentative production 2, the 3-butyleneglycol helps reducing the fermentative production cost, realize the high value development and use of biomass resource, and utilize renewable biomass development of resources petroleum chemicals substitute to help the sustainable development of society.
At present bibliographical information be used for 2, the substrate of 3-butyleneglycol fermentation production mainly contains (Syu MJ.Appl Microbiol Biotechnol, 2001,55:10-18 such as glucose, sucrose, molasses, glycerine; Qin et al.Chin J Chem Eng, 2006,14:132-136; Afschar et al.Appl Microbiol Biotechnol, 1991,34:582-585; Petrov K and PetrovaP, Appl Microbiol Biotechnol, DOI 10.1007/s00253-009-2004-x), 2 of patent report, the production of 3-butyleneglycol has: Chinese patent application numbers 200610113390.9 discloses a kind of plant straw fermentative production 2, method of 3-butyleneglycol utilized, liquid fermenting obtains 2 of 0.14～0.24g/L, the 3-butyleneglycol, solid state fermentation 2, the output of 3-butyleneglycol is at 0.026～0.067g/g siccative.Chinese patent application numbers 200810057041.9 discloses a kind of by preparing ethanol from wood fiber raw material and 2, and the method for 3-butyleneglycol can obtain 42.5g/L 2 in 72 hours, the 3-butyleneglycol.It is that fermenting raw materials produces 2 with the jerusalem artichoke that Chinese patent application numbers 200810011692.4 discloses a kind of, the method for 3-butyleneglycol, and having obtained final concentration is 2 of 49～105g/L, the 3-butyleneglycol.Chinese patent application numbers 200510011959.6 discloses a kind ofly produces 1 by crude starch material, ammediol and 2, the method for 3-butyleneglycol, this method is that fermentation obtains glycerol fermented broth through yeast cell by crude starch material earlier, after the glycerol fermented broth filtration treatment, produce 1, ammediol and 2 by the glycerol fermented broth of handling aerobic flow feeding after bacterium elder generation anaerobism again, the 3-butyleneglycol, but because the processing condition restriction, 1, ammediol is a primary product, 2,3-butyleneglycol concentration can reach 30.4g/L.
The starch based biomass material has source widely in China, and China is annual in recent years produces various starch above 1,000 ten thousand tons, has developed into world starch largest production state.Starch comprises W-Gum, potato starch, modified starch etc.Starch materials successfully has been used for the fermentative production of products such as ethanol, lactic acid, citric acid at present.Utilize starch materials direct fermentation to produce 2, the rarely seen report of 3-butyleneglycol.
Summary of the invention
The purpose of this invention is to provide a kind of starch materials fermentative production 2, the method for 3-butyleneglycol to realize the abundant development and use of starch materials, is produced high value hardware and software platform compound 2, the 3-butyleneglycol.
The starch materials that utilizes of the present invention produces 2, and the method for 3-butyleneglycol is characterized in that comprising the steps:
(1) starch materials is handled and the preparation of diastatic fermentation substratum step by step:
Be that 1～6: 1 ratio adds starch materials and is mixed into starch slurry in the tap water with water and starch materials mass ratio; Regulate starch slurry pH value 5.5～8.0, add α-Dian Fenmei in starch slurry, the α-Dian Fenmei enzyme concentration is 1～10U/g starch dry weight, under 90～110 ℃ of conditions, liquefies 0.5～4 hour, obtains liquefied starch; Liquefied starch pH value is adjusted in 3.5～6.0 backs adds saccharifying enzyme, the addition of saccharifying enzyme is 100～1000U/g starch dry weight, and saccharification is 0.5～6 hour under 50～70 ℃ of conditions, obtains starch saccharificating liquid, and its glucose concn scope is 60～510g/L; With 115 ℃ of sterilizations of saccharification liquid 20min, the cooling back is used to prepare the fermention medium that carries out the substep diastatic fermentation as carbon source;
Be suitable for carrying out the fermention medium of substep saccharification batch fermentation, prescription is: carbon source is a starch saccharificating liquid, and its consumption contains glucose amount with it and counts 100～120g/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Be suitable for carrying out the fermention medium of substep saccharification fed-batch fermentation, prescription is: carbon source is a starch saccharificating liquid, and its consumption contains glucose amount with it and counts 40～60g/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Above-mentioned starch materials is one or more any part by weight mixtures in Semen Maydis powder, W-Gum, Tapioca Starch, tapioca (flour), mealy potato, potato starch, the Ipomoea batatas dry powder;
(2) 5 liters of fermentor tank substep diastatic fermentations produce 2, the 3-butyleneglycol:
Inoculum size with volume ratio 5～10%, Klebsiella pneumonia (Klebsiella pneumoniae) SDMCCTCC M 208097 or Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 or Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 seed liquor of preparation are inoculated in 5 liters of fermentor tanks that 2～4 liters of substep diastatic fermentation substratum are housed, with leavening temperature is 30～40 ℃, mixing speed is 200～500rpm, and air flow 0.5～1.5vvm implements fermentation; In the fermenting process, add the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream
3PO
4Regulate the pH value, make it to be controlled at pH 5.5～6.5; And the glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; If carry out substep saccharification batch fermentation, when 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, get fermented liquid and separate, extract 2, the 3-butyleneglycol; If carry out substep saccharification fed-batch fermentation, when the detection glucose concn drops to 15～20g/L, add the starch saccharificating liquid of above-mentioned sterilization, keep in the fermented liquid glucose concn, when recording glucose consumption speed and be lower than 2g/ (Lh) at 15～60g/L, stop to add starch saccharificating liquid, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, getting fermented liquid separates, extracts 2, the 3-butyleneglycol;
Perhaps:
(1) preparation of starch materials processing and simultaneous saccharification and fermentation substratum:
Be that 1～6: 1 ratio adds starch materials and is mixed into starch slurry in the tap water with water and starch materials mass ratio; Regulate starch slurry pH value 5.5～8.0, add α-Dian Fenmei in starch slurry, the α-Dian Fenmei enzyme concentration is 1～10U/g starch dry weight, under 90～110 ℃ of conditions, liquefies 0.5～4 hour, obtains liquefied starch; With 115 ℃ of sterilizations of liquefied starch 20min, the cooling back is used to prepare the fermention medium that carries out simultaneous saccharification and fermentation as carbon source;
Be suitable for carrying out the fermention medium of synchronous saccharification batch fermentation, prescription is: carbon source is a liquefied starch, and its consumption is 250～600mL/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Be suitable for carrying out the fermention medium of synchronous saccharification fed-batch fermentation, prescription is: carbon source is a liquefied starch, and its consumption is 100～300mL/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Above-mentioned starch materials is one or more any part by weight mixtures in Semen Maydis powder, W-Gum, Tapioca Starch, tapioca (flour), mealy potato, potato starch, the Ipomoea batatas dry powder;
(2) 5 liters of fermentor tank simultaneous saccharification and fermentations produce 2, the 3-butyleneglycol:
Inoculum size with volume ratio 5～10%, Klebsiella pneumonia (Klebsiella pneumoniae) SDMCCTCC M 208097 or Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 or Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 seed liquor of preparation are inoculated in 5 liters of fermentor tanks that 2～4 liters of simultaneous saccharification and fermentation substratum are housed, add the saccharifying enzyme of filtration sterilization simultaneously, the addition of saccharifying enzyme is 100～1000U/g starch dry weight; With leavening temperature is 30～40 ℃, and mixing speed is 200～500rpm, and air flow 0.5～1.5vvm implements fermentation; In the fermenting process, add the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream
3PO
4Regulate the pH value, make it to be controlled at pH 5.5～6.5; And the glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; If carry out the synchronous saccharification batch fermentation, when the detection glucose concn drops to 15～20g/L, the saccharifying enzyme that adds filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, make in the fermented liquid glucose concn at 15～40g/L, when glucose concn no longer increases after adding saccharifying enzyme, stop to add saccharifying enzyme; Treat that glucose has consumed, 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, get fermented liquid and separate, extract 2, the 3-butyleneglycol; If carry out the synchronous saccharification fed-batch fermentation, when the detection glucose concn drops to 15～20g/L, add the liquefied starch of 200～800mL sterilization and add the saccharifying enzyme of filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, to keep in the fermented liquid glucose concn at 15～60g/L, when recording glucose consumption speed and be lower than 2g/ (Lh), stop to add liquefied starch and saccharifying enzyme, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, get fermented liquid and separate, extract 2, the 3-butyleneglycol.
The preferred Semen Maydis powder of above-mentioned starch materials, W-Gum, Tapioca Starch or tapioca (flour).
Above-mentioned amount of water and starch materials mass ratio preferred 1～4: 1.
Preferred 2～6U/g the starch of the enzyme concentration of above-mentioned α-Dian Fenmei dry weight.
In the above-mentioned substep diastatic fermentation process, the preferred 400～800U/g starch of the addition of saccharifying enzyme dry weight.
In the above-mentioned simultaneous saccharification and fermentation process, the preferred 200～400U/g starch of the addition of saccharifying enzyme dry weight.
Preferred 35～37 ℃ of above-mentioned leavening temperature, the preferred 300～400rpm of mixing speed, the preferred 0.8～1.2vvm of air flow.
The preparation method of above-mentioned Friedlanders bacillus (Klebsiella pneumoniae) SDM CCTCC M 208097 or Friedlanders bacillus (Klebsiella pneumoniae) CICC 10011 or Friedlanders bacillus (Klebsiella pneumoniae) ATCC 8724 seed liquor is: Friedlanders bacillus (Klebsiella pneumoniae) SDMCCTCC M 208097 or Friedlanders bacillus (Klebsiella pneumoniae) CICC 10011 or Friedlanders bacillus (Klebsiella pneumoniae) ATCC 8724 bacterial classifications of activation are received in the seed culture medium of sterilization; 30～37 ℃ of of At, 100～150rpm cultivates and obtained seed liquor in, 10～14 hours.
Wherein said seed culture based formulas is: glucose 40g/L, NH
4Cl 5g/L, K
2HPO
43H
2O 3g/L, KH
2PO
43g/L, Na
2SO
42g/L, MgSO
47H
2O 0.3g/L, ZnSO
47H
2O 0.2g/L.
Distinguishing feature of the present invention is:
1. the starch based biomass material in making full use of the natural resources comes direct fermentation production high value hardware and software platform compound 2, and the 3-butyleneglycol can improve the utilising efficiency of biomass resource, produces higher social benefit.And utilize renewable resources to replace petrochemical industry resource production high value chemical, help the sustainable development of economic society.Used starch based biomass material wide material sources, cheap, reprocessing cycle is rapid, and environmental friendliness can significantly reduce the fermentative production cost.
2. the present invention has obtained the zymotechnique of optimizing by the research to aspects such as water addition ratio, α-Dian Fenmei dosage, saccharifying enzyme dosage, enzyme treatment temp and treatment time, fermentation time, fermentation condition optimization.
3. by comparing, adopt simultaneous saccharification and fermentation more to have superiority: simultaneous saccharification and fermentation has omitted saccharification stage, energy efficient; Saccharification and fermentation are carried out in same reactor, save facility investment; Saccharification and fermentation are carried out simultaneously in addition, and the glucose that saccharification produces is very fast to be utilized by thalline, keeps lower level, can avoid the substrate inhibition, have eliminated the lag period of thalline to high concentration substrate simultaneously, have shortened fermentation period, have improved production efficiency.
4. the present invention has realized effective utilization of starch based biomass material, number is that 200510011959.6 disclosed methods are compared with Chinese patent application, and the inventive method has been avoided and used 2 kinds of bacterium to ferment, and need not through complicated fermenting process.Anaerobism prepared glycerol fermented broth after Chinese patent application number 200510011959.6 methods of announcing were to use yeast elder generation aerobic, after the glycerol fermented broth filtration treatment, the aerobic flow feeding produces 1 after utilizing bacterium by glycerol fermented broth elder generation anaerobism again, ammediol and 2, the 3-butyleneglycol, 1, ammediol is a primary product, 2,3-butyleneglycol concentration can reach 30.4g/L.The present invention has simplified the fermentation operation flow process, utilizes starch materials to obtain 2 of high yield through fermentation using bacteria, the 3-butyleneglycol.By substep diastatic fermentation or simultaneous saccharification and fermentation, 2, the ultimate density of 3-butyleneglycol reaches 43～112g/L, acetoin and 2, the ultimate density of 3-butyleneglycol reaches 45～121g/L, has great promotion and application prospect.
Embodiment
The present invention is further elaborated below in conjunction with embodiment, but be not limited only to this.
General explanation:
The selected bacterial strain of the present invention is Klebsiella pneumonia (Klebsiella pneumoniae) SDM CCTCC M208097 or Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 or Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724.Wherein, Klebsiella pneumonia (Klebsiella pneumoniae) SDMCCTCC M 208097 screens and is deposited in Chinese typical culture collection center by Shandong University (laboratory, the present application people place); Klebsiella pneumonia CICC 10011 can be buied by Chinese industrial microbial strains management preservation center; Klebsiella pneumonia ATCC 8724 can be buied by the biological product of USS collecting center.
The determination of glucose method that relates in the inventive method is: fermented liquid dilution back is centrifugal, adopts bio-sensing analyser SBA-40C (Shandong Province academy sciences Biology Research Institute) to measure.Measuring principle is measured glucose content for utilizing immobilized glucose oxidase film specificity.
Relate in the inventive method 2, the measuring method of 3-butyleneglycol is: VARIAN CP-3380 gas chromatograph detects.Butylacetate is an extraction agent, and volume ratio extraction in 1: 1 is got upper organic phase and detected.The concrete condition of measuring is: flame ion monitor (FID) temperature is 280 ℃, and the sampler temperature is 220 ℃, and the kapillary column temperature is to be warmed up to 180 ℃ from 50 ℃, 20 ℃/min of heat-up rate, and carrier gas is a nitrogen.Utilize 2,3-butyleneglycol standard substance (German Sigma-Aldrich company, article No.: 361461) make typical curve, calculate in the fermented liquid 2 according to typical curve again, the content of 3-butyleneglycol.
Embodiment 1: utilizing Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 is that raw material substep saccharification fed-batch fermentation in 5 liters of fermentor tanks produces 2, the 3-butyleneglycol with the W-Gum
(1) W-Gum is handled and the preparation of saccharification fed-batch fermentation substratum step by step:
The 1200g W-Gum being added water be modulated into starch slurry and be settled to 2.5L, regulate pH to 5.5, is that 10U/g starch dry weight adds α-Dian Fenmei by enzyme concentration, and boiling is 2 hours under 95 ℃ of conditions, obtains the W-Gum liquefier; The W-Gum liquefier is cooled to 55 ℃, regulate the pH to 4.0 of liquefier, by enzyme concentration is that 1000U/g starch dry weight adds saccharifying enzyme, under 55 ℃ of conditions, handled 2 hours, obtain W-Gum saccharification liquid, recording glucose concn is 452g/L, and with 115 ℃ of sterilizations of saccharification liquid 20 minutes, the cooling back was used to prepare the fermention medium that carries out substep saccharification fed-batch fermentation as carbon source;
Be suitable for carrying out the fermention medium of substep saccharification fed-batch fermentation, prescription is: carbon source is a W-Gum saccharification liquid, and its consumption contains glucose amount with it and counts 45g/L; All the other compositions are: (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.5,115 ℃ of sterilizations 20 minutes;
(2) 5 liters of fermentor tank substep saccharification fed-batch fermentations produce 2, the 3-butyleneglycol:
Under the aseptic condition be the fermention medium that 5% inoculum size inserts sterilization by volume with cultured Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 seed liquor, fermentor tank initial loading liquid measure is 2L, with leavening temperature is 35 ℃, mixing speed is 350rpm, and air flow is that 1.2vvm ferments; The fermentation initial pH value is 6.5, in the fermenting process, when the pH value is reduced to 6.0, adds the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream
3PO
4Regulate the pH value, make it to be controlled at pH 6.0; Glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; In the fermenting process, when the detection glucose concn drops to 15～20g/L, add the W-Gum saccharification liquid of 200～400mL sterilization, keep in the fermented liquid glucose concn at 15～60g/L, when recording glucose consumption speed and be lower than 2g/ (Lh), stop to add W-Gum saccharification liquid, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, final fermentation time is 66 hours; After fermentation finishes, 2, the concentration of 3-butyleneglycol is 107.87g/L, acetoin and 2, the total content of 3-butyleneglycol is 114.69g/L, production efficiency is 1.74g/ (Lh).
Embodiment 2: utilizing Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 is that raw material synchronous saccharification fed-batch fermentation in 5 liters of fermentor tanks produces 2, the 3-butyleneglycol with the W-Gum
(1) preparation of W-Gum processing and synchronous saccharification fed-batch fermentation substratum:
The 1200g W-Gum is added water to be modulated into starch slurry and to be settled to 2.5L, regulate pH to 6.5, by enzyme concentration is that 2U/g starch dry weight adds α-Dian Fenmei, boiling is 4 hours under 90 ℃ of conditions, obtain the W-Gum liquefier, sterilized 20 minutes for 115 ℃, the cooling back is used to prepare the fermention medium that carries out the synchronous saccharification fed-batch fermentation as carbon source;
Be suitable for carrying out the fermention medium of synchronous saccharification fed-batch fermentation, prescription is: carbon source is the W-Gum liquefier, and its consumption is 150mL/L; All the other compositions are: (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0,115 ℃ of sterilizations 20 minutes;
(2) 5 liters of fermentor tank synchronous saccharification fed-batch fermentations produce 2, the 3-butyleneglycol:
Under the aseptic condition be the fermention medium that 10% inoculum size inserts sterilization by volume with cultured Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 seed liquor, add the saccharifying enzyme of filtration sterilization simultaneously, the saccharifying enzyme addition is a 300U/g starch dry weight; Fermentor tank initial loading liquid measure is 2L, is 37 ℃ with leavening temperature, and mixing speed is 450rpm, and air flow is that 1.2vvm ferments; The fermentation initial pH value is 6.0, adds the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream in the fermenting process
3PO
4Regulate the pH value and make it to maintain pH 6.0; Glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; In the fermenting process, when the detection glucose concn drops to 15～20g/L, add the W-Gum liquefier of 200～400mL sterilization and add the saccharifying enzyme of filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, to keep in the fermented liquid glucose concn at 15～60g/L, when recording glucose consumption speed and be lower than 2g/ (Lh), stop to add W-Gum liquefier and saccharifying enzyme, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, fermentation time is 58 hours; After fermentation finishes, 2, the concentration of 3-butyleneglycol is 110.63g/L, acetoin and 2, the total content of 3-butyleneglycol is 120.87g/L, production efficiency is 2.08g/ (Lh).Compare with embodiment 1, the W-Gum simultaneous saccharification and fermentation has shortened fermentation period, acetoin and 2, and the production efficiency of 3-butyleneglycol improves a lot.
Embodiment 3: utilizing Klebsiella pneumonia (Klebsiella pneumoniae) SDM CCTCC M 208097 is that raw material synchronous saccharification fed-batch fermentation in 5 liters of fermentor tanks produces 2, the 3-butyleneglycol with the Tapioca Starch
(1) preparation of Tapioca Starch processing and synchronous saccharification fed-batch fermentation substratum:
The 540g Tapioca Starch is added water to be modulated into starch slurry and to be settled to 1.2L, regulate pH to 6.5, by enzyme concentration is that 5U/g starch dry weight adds α-Dian Fenmei, boiling is 1 hour under 90 ℃ of conditions, obtain the Tapioca Starch liquefier, sterilized 20 minutes for 115 ℃, the cooling back is used to prepare the fermention medium that carries out the synchronous saccharification fed-batch fermentation as carbon source;
Be suitable for carrying out the fermention medium of synchronous saccharification fed-batch fermentation, prescription is: carbon source is the Tapioca Starch liquefier, and its consumption is 170mL/L; All the other compositions are: (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0,115 ℃ of sterilizations 20 minutes;
(2) 5 liters of fermentor tank synchronous saccharification fed-batch fermentations produce 2, the 3-butyleneglycol:
Under the aseptic condition be the fermention medium that 10% inoculum size inserts sterilization by volume with cultured Klebsiella pneumonia (Klebsiella pneumoniae) SDM CCTCC M208097 seed liquor, add the saccharifying enzyme of filtration sterilization simultaneously, the saccharifying enzyme addition is a 500U/g starch dry weight; Fermentor tank initial loading liquid measure is 1.2L, is 35 ℃ with leavening temperature, and mixing speed is 400rpm, and air flow is that 0.6vvm ferments; The fermentation initial pH value is 6.0, adds the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream in the fermenting process
3PO
4Regulate the pH value and make it to maintain pH6.0; Glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; In the fermenting process, when the detection glucose concn drops to 15～20g/L, add the Tapioca Starch liquefier of 100～200mL sterilization and add the saccharifying enzyme of filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, to keep in the fermented liquid glucose concn at 15～60g/L, when recording glucose consumption speed and be lower than 2g/ (Lh), stop to add Tapioca Starch liquefier and saccharifying enzyme, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, fermentation time is 60 hours; After fermentation finishes, 2, the concentration of 3-butyleneglycol is 62.73g/L, acetoin and 2, the total content of 3-butyleneglycol is 65.10g/L, production efficiency is 1.09g/ (Lh).
Embodiment 4: utilizing Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 is that raw material synchronous saccharification batch fermentation in 5 liters of fermentor tanks produces 2, the 3-butyleneglycol with the tapioca (flour)
(1) preparation of tapioca (flour) processing and synchronous saccharification batch fermentation substratum:
The 350g tapioca (flour) is added water to be modulated into starch slurry and to be settled to 1L, regulate pH to 6.0, by enzyme concentration is that 6U/g starch dry weight adds α-Dian Fenmei, boiling is 1 hour under 95 ℃ of conditions, obtain the tapioca (flour) liquefier, sterilized 20 minutes for 115 ℃, the cooling back is used to prepare the fermention medium that carries out the synchronous saccharification batch fermentation as carbon source;
Be suitable for carrying out the fermention medium of synchronous saccharification batch fermentation, prescription is: carbon source is a liquefied starch, and its consumption is 500mL/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.5,115 ℃ of sterilizations 20 minutes;
(2) 5 liters of fermentor tank synchronous saccharification batch fermentations produce 2, the 3-butyleneglycol:
Under the aseptic condition be the fermention medium that 5% inoculum size inserts sterilization by volume with cultured Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 seed liquor, add the saccharifying enzyme of filtration sterilization simultaneously, the saccharifying enzyme addition is a 200U/g starch dry weight; The total liquid amount of fermentor tank is 2L, is 37 ℃ with leavening temperature, and mixing speed is 350rpm, and air flow is that 1.0vvm ferments; The fermentation initial pH value is 6.5, in the fermenting process, when the pH value is reduced to 6.0, adds the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream in the fermenting process
3PO
4Regulate the pH value, make it to be controlled at pH 6.0; Glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; When the detection glucose concn drops to 15～20g/L, the saccharifying enzyme that adds filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, to keep in the fermented liquid glucose concn at 15～40g/L, when glucose concn no longer increases after adding saccharifying enzyme, stop to add saccharifying enzyme, treat that glucose has consumed, 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, fermentation time is 40 hours; After fermentation finishes, 2, the concentration of 3-butyleneglycol is 69.48g/L, acetoin and 2, the total content of 3-butyleneglycol is 72.32g/L, production efficiency is 1.81g/ (Lh).
Embodiment 5: utilizing Klebsiella pneumonia (Klebsiella pneumoniae) SDM CCTCC M 208097 is that raw material substep saccharification fed-batch fermentation in 5 liters of fermentor tanks produces 2, the 3-butyleneglycol with Ipomoea batatas dry powder
(1) Ipomoea batatas dry powder is handled and the preparation of saccharification fed-batch fermentation substratum step by step:
1000g Ipomoea batatas dry powder being added water be modulated into starch slurry and be settled to 2L, regulate pH to 5.5, is that 5U/g starch dry weight adds α-Dian Fenmei by enzyme concentration, and boiling is 1 hour under 94 ℃ of conditions, obtains Ipomoea batatas dry powder liquefier; Ipomoea batatas dry powder liquefier is cooled to 60 ℃, regulate the pH to 4.3 of liquefier, by enzyme concentration is that 300U/g starch dry weight adds saccharifying enzyme, under 60 ℃ of conditions, handled 6 hours, obtain dried sweet potato pulverized sugar liquid, recording glucose concn is 312g/L, and with 115 ℃ of sterilizations of saccharification liquid 20 minutes, the cooling back was used to prepare the fermention medium that carries out substep saccharification fed-batch fermentation as carbon source;
Be suitable for carrying out the fermention medium of substep saccharification fed-batch fermentation, prescription is: carbon source is a dried sweet potato pulverized sugar liquid, and its consumption contains glucose amount with it and counts 38g/L; All the other compositions are: (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0,115 ℃ of sterilizations 20 minutes;
(2) 5 liters of fermentor tank substep saccharification fed-batch fermentations produce 2, the 3-butyleneglycol:
Under the aseptic condition be the fermention medium that 10% inoculum size inserts sterilization by volume with cultured Klebsiella pneumonia (Klebsiella pneumoniae) SDM CCTCC M208097 seed liquor, fermentor tank initial loading liquid measure is 2.2L, with leavening temperature is 35 ℃, mixing speed is 300rpm, and air flow is that 1.2vvm ferments; The fermentation initial pH value is 6.0, adds the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream in the fermenting process
3PO
4Regulate the pH value and make it to maintain pH 6.0; Glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; In the fermenting process, when the detection glucose concn drops to 15～20g/L, add the dried sweet potato pulverized sugar liquid of 200～500mL sterilization, keep in the fermented liquid glucose concn at 15～60g/L, when recording glucose consumption speed and be lower than 2g/ (Lh), stop to add dried sweet potato pulverized sugar liquid, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, final fermentation time is 44 hours; After fermentation finishes, 2, the concentration of 3-butyleneglycol is 50.28g/L, acetoin and 2, the total content of 3-butyleneglycol is 56.94g/L, production efficiency is 1.29g/ (Lh).
Embodiment 6: utilizing Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 is that raw material synchronous saccharification batch fermentation in 5 liters of fermentor tanks produces 2, the 3-butyleneglycol with the potato starch
(1) preparation of potato starch processing and synchronous saccharification batch fermentation substratum:
The 400g potato starch is added water to be modulated into starch slurry and to be settled to 1L, regulate pH 6.5, by enzyme concentration is that 8U/g starch dry weight adds α-Dian Fenmei, boiling is 1.5 hours under 95 ℃ of conditions, obtain the potato starch liquefier, sterilized 20 minutes for 115 ℃, the cooling back is used to prepare the fermention medium that carries out the synchronous saccharification batch fermentation as carbon source;
Be suitable for carrying out the fermention medium of synchronous saccharification batch fermentation, prescription is: carbon source is a liquefied starch, and its consumption is 500mL/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 7.0,115 ℃ of sterilizations 20 minutes;
(2) 5 liters of fermentor tank synchronous saccharification batch fermentations produce 2, the 3-butyleneglycol:
Under the aseptic condition be the fermention medium that 10% inoculum size inserts sterilization by volume with cultured Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 seed liquor, add the saccharifying enzyme of filtration sterilization simultaneously, the saccharifying enzyme addition is a 300U/g starch dry weight; The total liquid amount of fermentor tank is 2L, is 37 ℃ with leavening temperature, and mixing speed is 300rpm, and air flow is that 1.0vvm ferments; The fermentation initial pH value is 7.0, in the fermenting process, when the pH value is reduced to 5.5, adds the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream in the fermenting process
3PO
4Regulate the pH value, make it to be controlled at pH 5.5; Glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; When the detection glucose concn drops to 15～20g/L, the saccharifying enzyme that adds filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, to keep in the fermented liquid glucose concn at 15～40g/L, when glucose concn no longer increases after adding saccharifying enzyme, stop to add saccharifying enzyme, treat that glucose has consumed, 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, fermentation time is 52 hours; After fermentation finishes, 2, the concentration of 3-butyleneglycol is 81.91g/L, acetoin and 2, the total content of 3-butyleneglycol is 86.18g/L, production efficiency is 1.66g/ (Lh).
Claims (7)
1. one kind is utilized starch materials to produce 2, and the method for 3-butyleneglycol is characterized in that comprising the steps:
(1) starch materials is handled and the preparation of diastatic fermentation substratum step by step:
Be that 1～6: 1 ratio adds starch materials and is mixed into starch slurry in the tap water with water and starch materials mass ratio; Regulate starch slurry pH value 5.5～8.0, add α-Dian Fenmei in starch slurry, the α-Dian Fenmei enzyme concentration is 1～10U/g starch dry weight, under 90～110 ℃ of conditions, liquefies 0.5～4 hour, obtains liquefied starch; Liquefied starch pH value is adjusted in 3.5～6.0 backs adds saccharifying enzyme, the addition of saccharifying enzyme is 100～1000U/g starch dry weight, and saccharification is 0.5～6 hour under 50～70 ℃ of conditions, obtains starch saccharificating liquid, and its glucose concn scope is 60～510g/L; With 115 ℃ of sterilizations of saccharification liquid 20min, the cooling back is used to prepare the fermention medium that carries out the substep diastatic fermentation as carbon source;
Be suitable for carrying out the fermention medium of substep saccharification batch fermentation, prescription is: carbon source is a starch saccharificating liquid, and its consumption contains glucose amount with it and counts 100～120g/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Be suitable for carrying out the fermention medium of substep saccharification fed-batch fermentation, prescription is: carbon source is a starch saccharificating liquid, and its consumption contains glucose amount with it and counts 40～60g/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Above-mentioned starch materials is one or more any part by weight mixtures in Semen Maydis powder, W-Gum, Tapioca Starch, tapioca (flour), mealy potato, potato starch, the Ipomoea batatas dry powder;
(2) 5 liters of fermentor tank substep diastatic fermentations produce 2, the 3-butyleneglycol:
Inoculum size with volume ratio 5～10%, Klebsiella pneumonia (Klebsiella pneumoniae) SDMCCTCC M 208097 or Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 or Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 seed liquor of preparation are inoculated in 5 liters of fermentor tanks that 2～4 liters of substep diastatic fermentation substratum are housed, with leavening temperature is 30～40 ℃, mixing speed is 200～500rpm, and air flow 0.5～1.5vvm implements fermentation; In the fermenting process, add the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream
3PO
4Regulate the pH value, make it to be controlled at pH 5.5～6.5; And the glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; If carry out substep saccharification batch fermentation, when 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, get fermented liquid and separate, extract 2, the 3-butyleneglycol; If carry out substep saccharification fed-batch fermentation, when the detection glucose concn drops to 15～20g/L, add the starch saccharificating liquid of above-mentioned sterilization, keep in the fermented liquid glucose concn, when recording glucose consumption speed and be lower than 2g/ (Lh) at 15～60g/L, stop to add starch saccharificating liquid, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, getting fermented liquid separates, extracts 2, the 3-butyleneglycol;
Perhaps:
(1) preparation of starch materials processing and simultaneous saccharification and fermentation substratum:
Be that 1～6: 1 ratio adds starch materials and is mixed into starch slurry in the tap water with water and starch materials mass ratio; Regulate starch slurry pH value 5.5～8.0, add α-Dian Fenmei in starch slurry, the α-Dian Fenmei enzyme concentration is 1～10U/g starch dry weight, under 90～110 ℃ of conditions, liquefies 0.5～4 hour, obtains liquefied starch; With 115 ℃ of sterilizations of liquefied starch 20min, the cooling back is used to prepare the fermention medium that carries out simultaneous saccharification and fermentation as carbon source;
Be suitable for carrying out the fermention medium of synchronous saccharification batch fermentation, prescription is: carbon source is a liquefied starch, and its consumption is 250～600mL/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Be suitable for carrying out the fermention medium of synchronous saccharification fed-batch fermentation, prescription is: carbon source is a liquefied starch, and its consumption is 100～300mL/L, (NH
4)
2SO
416g/L, K
2HPO
43H
2O 4g/L, KH
2PO
44g/L, NaCl 2g/L, CaCl
20.4g/L, MgSO
47H
2O 0.4g/L, ZnSO
47H
2O 0.4g/L, potassium acetate 4g/L, trisodium citrate 4g/L; The pH value transfers to 6.0～7.0;
Above-mentioned starch materials is one or more any part by weight mixtures in Semen Maydis powder, W-Gum, Tapioca Starch, tapioca (flour), mealy potato, potato starch, the Ipomoea batatas dry powder;
(2) 5 liters of fermentor tank simultaneous saccharification and fermentations produce 2, the 3-butyleneglycol:
Inoculum size with volume ratio 5～10%, Klebsiella pneumonia (Klebsiella pneumoniae) SDMCCTCC M 208097 or Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 or Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 seed liquor of preparation are inoculated in 5 liters of fermentor tanks that 2～4 liters of simultaneous saccharification and fermentation substratum are housed, add the saccharifying enzyme of filtration sterilization simultaneously, the addition of saccharifying enzyme is 100～1000U/g starch dry weight; With leavening temperature is 30～40 ℃, and mixing speed is 200～500rpm, and air flow 0.5～1.5vvm implements fermentation; In the fermenting process, add the KOH of 4～6M or the H of 3～6M by the related control of fermentor tank peristaltic pump stream
3PO
4Regulate the pH value, make it to be controlled at pH 5.5～6.5; And the glucose content and 2 in 3～4 hours sampling and measuring fermented liquids, 3-butyleneglycol concentration; If carry out the synchronous saccharification batch fermentation, when the detection glucose concn drops to 15～20g/L, the saccharifying enzyme that adds filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, make in the fermented liquid glucose concn at 15～40g/L, when glucose concn no longer increases after adding saccharifying enzyme, stop to add saccharifying enzyme; Treat that glucose has consumed, 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, get fermented liquid and separate, extract 2, the 3-butyleneglycol; If carry out the synchronous saccharification fed-batch fermentation, when the detection glucose concn drops to 15～20g/L, add the liquefied starch of 200～800mL sterilization and add the saccharifying enzyme of filtration sterilization, the addition of saccharifying enzyme is 100～500U/g starch dry weight, to keep in the fermented liquid glucose concn at 15～60g/L, when recording glucose consumption speed and be lower than 2g/ (Lh), stop to add liquefied starch and saccharifying enzyme, treat 2, when 3-butyleneglycol concentration no longer rises, finish fermentation, get fermented liquid and separate, extract 2, the 3-butyleneglycol.
2. utilize starch materials to produce 2 according to claim 1, the method for 3-butyleneglycol is characterized in that: described starch materials is Semen Maydis powder, W-Gum, Tapioca Starch or tapioca (flour).
3. utilize starch materials to produce 2 according to claim 1, the method for 3-butyleneglycol is characterized in that: described amount of water and starch materials mass ratio are 1～4: 1.
4. utilize starch materials to produce 2 according to claim 1, the method for 3-butyleneglycol is characterized in that: the enzyme concentration of described α-Dian Fenmei is 2～6U/g starch dry weight.
5. utilize starch materials to produce 2 according to claim 1, the method for 3-butyleneglycol is characterized in that: in the described substep diastatic fermentation process, the addition of saccharifying enzyme is 400～800U/g starch dry weight; In the described simultaneous saccharification and fermentation process, the addition of saccharifying enzyme is 200～400U/g starch dry weight.
6. utilize starch materials to produce 2 according to claim 1, the method for 3-butyleneglycol is characterized in that: described leavening temperature is 35～37 ℃, and mixing speed is 300～400rpm, and air flow is 0.8～1.2vvm.
7. utilize starch materials to produce 2 according to claim 1, the method of 3-butyleneglycol, it is characterized in that: the preparation method of described Klebsiella pneumonia (Klebsiella pneumoniae) SDM CCTCC M 208097 or Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 or Klebsiella pneumonia (Klebsiella pneumoniae) ATCC8724 seed liquor is: activatory Klebsiella pneumonia (Klebsiella pneumoniae) SDM CCTCCM 208097 or Klebsiella pneumonia (Klebsiella pneumoniae) CICC 10011 or Klebsiella pneumonia (Klebsiella pneumoniae) ATCC 8724 bacterial classifications are received in the seed culture medium of sterilization, at 30～37 ℃, 100～150rpm cultivates and obtained seed liquor in 10～14 hours;
Wherein, described seed culture based formulas is: glucose 40g/L, NH
4Cl 5g/L, K
2HPO
43H
2O 3g/L, KH
2PO
43g/L, Na
2SO
42g/L, MgSO
47H
2O 0.3g/L, ZnSO
47H
2O 0.2g/L.
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| CNA2009100154009A CN101565721A (en) | 2009-06-04 | 2009-06-04 | Method for producing 2,3-butanediol by using starch raw materials |
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| CNA2009100154009A CN101565721A (en) | 2009-06-04 | 2009-06-04 | Method for producing 2,3-butanediol by using starch raw materials |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101942487A (en) * | 2010-10-21 | 2011-01-12 | 江南大学 | Method for fermenting and preparing citric acid by adding saccharifying enzyme |
| CN102260717A (en) * | 2011-06-16 | 2011-11-30 | 山东柠檬生化有限公司 | Method for producing citric acid by fermentation |
| CN104726509A (en) * | 2015-02-09 | 2015-06-24 | 苏州科技学院 | Method for producing epsilon-polylysine through fermentation of cassava starch |
| CN104894171A (en) * | 2014-03-06 | 2015-09-09 | 广西科学院 | Method for producing (R,R)-2,3-butylene glycol by fermenting raw material cassava starch |
| CN105624213A (en) * | 2014-10-28 | 2016-06-01 | 中国石油化工股份有限公司 | Method for preparing 2,3-butanediol (BDO) by adopting microalgae as raw material |
| CN105624212A (en) * | 2014-10-28 | 2016-06-01 | 中国石油化工股份有限公司 | Method for preparing 2,3-butanediol (BDO) by adopting microalgae as raw material |
-
2009
- 2009-06-04 CN CNA2009100154009A patent/CN101565721A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101942487A (en) * | 2010-10-21 | 2011-01-12 | 江南大学 | Method for fermenting and preparing citric acid by adding saccharifying enzyme |
| CN102260717A (en) * | 2011-06-16 | 2011-11-30 | 山东柠檬生化有限公司 | Method for producing citric acid by fermentation |
| CN104894171A (en) * | 2014-03-06 | 2015-09-09 | 广西科学院 | Method for producing (R,R)-2,3-butylene glycol by fermenting raw material cassava starch |
| CN105624213A (en) * | 2014-10-28 | 2016-06-01 | 中国石油化工股份有限公司 | Method for preparing 2,3-butanediol (BDO) by adopting microalgae as raw material |
| CN105624212A (en) * | 2014-10-28 | 2016-06-01 | 中国石油化工股份有限公司 | Method for preparing 2,3-butanediol (BDO) by adopting microalgae as raw material |
| CN105624212B (en) * | 2014-10-28 | 2019-06-18 | 中国石油化工股份有限公司 | A method of 2,3- butanediol is produced by raw material of microalgae |
| CN105624213B (en) * | 2014-10-28 | 2019-08-06 | 中国石油化工股份有限公司 | A method of 2,3- butanediol is produced using microalgae for raw material |
| CN104726509A (en) * | 2015-02-09 | 2015-06-24 | 苏州科技学院 | Method for producing epsilon-polylysine through fermentation of cassava starch |
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