CN102517343A - Method for enhancing production intensity of producing itaconic acid by fermentation method - Google Patents
Method for enhancing production intensity of producing itaconic acid by fermentation method Download PDFInfo
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- CN102517343A CN102517343A CN2011104193368A CN201110419336A CN102517343A CN 102517343 A CN102517343 A CN 102517343A CN 2011104193368 A CN2011104193368 A CN 2011104193368A CN 201110419336 A CN201110419336 A CN 201110419336A CN 102517343 A CN102517343 A CN 102517343A
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- fermentation
- itaconic acid
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- production intensity
- culture medium
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for enhancing the production intensity of producing an itaconic acid by a fermentation method, which belongs to the technical field of fermentation engineering. According to the method for enhancing the production intensity of producing the itaconic acid by the fermentation method, fermentation optimization is performed by taking Aspergillus terreus FMME 033 as a production strain for producing the itaconic acid by utilizing a single-factor experiment method and an orthogonal experiment method so as to determine the inoculation cell age to be 36h and the fermentation period to be 48h, and determine the best compositions of a seed culture medium and a fermentation culture medium. According to the method for enhancing the production intensity of producing the itaconic acid by the fermentation method, the production intensity of the itaconic acid is effectively enhanced to 1.35g/(L h), and a new reference is provided for producing the itaconic acid by the fermentation method.
Description
Technical field
The present invention relates to a kind of method that fermentation method produces the methylene-succinic acid production intensity that improves, belong to field of fermentation engineering.
Background technology
Methylene-succinic acid (itaconic acid); Have another name called methylene-succinic acid; It is the unsaturated very widely binary organic acid of a kind of purposes; Can find out that by structural formula a unsaturated double-bond and two active carboxyls are arranged in the molecule, and carbon-carbon double bond and a carbonyl be conjugate relation, therefore have very strong reactive behavior.Utilize its two carboxyls, can be used as the monomer of unsaturated polyester; Utilize its two keys of C-C; Can with multiple substituted olefine copolymerization; Generation has the superpolymer of hydroxyl functional, makes it have widely in man-made fiber, synthetic resins, plastics, rubber, medicine, coating, tensio-active agent, water treatment, oil dope and medicine and other fields and uses.The Chemicals that methylene-succinic acid is earned foreign exchange as a kind of high-tech, high profit, height are described as the jewel on the imperial crown in the organic acid field, and development prospect is good.
The methylene-succinic acid working method has two types of fermentation method and chemical synthesiss, because the fermentative Production technical maturity, cost is lower etc., and advantage becomes the method that generally adopts both at home and abroad.After the wide down open country of nineteen twenty-nine Japan scholar wood finds that first mikrobe has the ability of secretion methylene-succinic acid; It is very active that the related microorganism fermenting carbohydrate is produced the research of methylene-succinic acid, reported that having the mikrobe that produces the methylene-succinic acid ability had candiyeast, rhodotorula, charles's mould, black mold P1, terreus and methylene-succinic acid aspergillus.The methylene-succinic acid industrial producing strain mainly is the terreus and the methylene-succinic acid aspergillus that belongs to the Aspergillus amstelodami crowd that belongs to the terreus crowd.
Fungi fermentation is produced the methylene-succinic acid problem that the ubiquity cycle is long in suitability for industrialized production and fundamental research, production intensity is lower.How to shorten fermentation period, improving production intensity does not still have good solution at present.
Summary of the invention
The purpose of this invention is to provide a kind of method that improves the obtained by producing itaconic acid by fermentation production intensity.
Technical scheme of the present invention: the terreus (Aspergillus terreus FMME 033) that uses screening under laboratory condition, utilizes single factor and orthogonal experiment method that substratum and culture condition are carried out shake flask fermentation optimization for producing bacterial strain.
Bacterial classification: the terreus of the product methylene-succinic acid of laboratory screening.
Beneficial effect of the present invention: utilize the present invention can effectively improve the methylene-succinic acid production intensity, shorten fermentation period.The output of methylene-succinic acid is 65g/L, and the production cycle is 48h, and production intensity is 1.35g/ (Lh), and effect is remarkable.
The present invention shortens fermentation period and also improves production intensity through a kind of strategy that utilizes microbial fermentation to produce methylene-succinic acid is provided, and obtained by producing itaconic acid by fermentation is from now on had certain directive significance.
Embodiment
Control Example: the used bacterial classification of the present invention is the terreus (Aspergillus terreus FMME 033) of laboratory screening.
7d is cultivated in the switching inclined-plane in 37 ℃ of incubators earlier, treats to wash with SPSS after the spore maturation, and spore suspension is adjusted spore concentration OD
560Be that 1.2 backs are inserted in the basic seed culture medium by 1%, 37 ℃ of temperature, cultivate 36h under the rotating speed 200rpm condition, it is 100mL that 500mL shakes bottled liquid measure, basic seed culture medium is carried out fermentation optimization confirm the best composition of substratum; From optimum seed medium, be forwarded to basic fermention medium, cultivate 120h by 10% inoculum size, 37 ℃ of temperature, it is 50mL that shaking speed 200rpm, 500mL shake bottled liquid measure, and fermention medium is carried out the best composition that fermentation optimization is confirmed fermention medium.
Inclined-plane seed solid medium (g/L):
Fructus Hordei Germinatus extract 26.9, glucose 20, peptone 1, agar 20, pH nature.
Basis seed culture medium (g/L):
Glucose 45, (NH
4)
2SO
42.67, MgSO
47H
2O 4, steeping water 1, initial pH 2.0.
Basis fermention medium (g/L):
Glucose 150, (NH
4)
2SO
42.67, MgSO
47H
2O 5, steeping water 1.5, initial pH 2.0.
Optimum seed medium (g/L): glucose 45, (NH
4)
2SO
45, MgSO
47H
2O 1, KH
2PO
41, FeSO
47H
2O0.04, ZnSO
47H
2O 0.03, steeping water 1, initial pH 3.0.
Optimal compositions of fermentation medium (g/L): glucose 100, (NH
4)
2SO
44, MgSO
47H
2O 1, FeSO
47H
2O 0.04, ZnSO
47H
2O 0.05, steeping water 3, initial pH 3.0.
Embodiment 1: the same reference examples of fermentation condition, the method for utilizing single factor and orthogonal experiment is optimized initial pH, carbon source, nitrogenous source, the inorganic salt of seed culture medium.
Find: the cell age of inoculation is constant before and after optimizing, but dry cell weight is increased to 7.4g/L by initial 2.3g/L, increases to original 3.2 times.
Embodiment 2: the same reference examples of fermentation condition, the method for utilizing single factor and orthogonal experiment is optimized initial pH, carbon source, nitrogenous source, the inorganic salt of fermention medium.
Find: 1. optimizing the secondary fermentation cycle shortens to 48h by 120h, shortens to original 40%; 2. dry cell weight is increased to 9.8g/L by 3.9g/L in the fermented liquid, rises to original 2.5 times; 3. fermented liquid underpants health acid content is increased to 65g/L, and production intensity is 1.35g/ (Lh).
Claims (2)
1. method that improves the obtained by producing itaconic acid by fermentation production intensity is characterized in that:
(1) terreus (Aspergillus terreus FMME 033) that uses screening is for producing bacterial strain;
(2) substratum consists of:
Seed culture medium (g/L):
Glucose 45, (NH
4)
2SO
45, MgSO
47H
2O 1, KH
2PO
41, FeSO
47H
2O 0.04, ZnSO
47H
2O 0.03, steeping water 1, initial pH 3.0;
Fermention medium (g/L):
Glucose 100, (NH
4)
2SO
44, MgSO
47H
2O 1, FeSO
47H
2O 0.04, ZnSO
47H
2O 0.05, steeping water 3, initial pH 3.0;
(3) culture condition is:
Culture temperature is 37 ℃, 200rpm, and shaking bottled liquid measure is that seed culture medium is 100mL/500mL, the spore inoculating amount is adjustment spore concentration OD
560Be that seed culture medium is inserted by 1% in 1.2 backs, fermention medium is 50mL/500mL, and the fermentation culture inoculum size is 10%.
2. method according to claim 1 is characterized in that fermentation period is 48h, and production intensity is 1.35g/ (Lh).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104193368A CN102517343A (en) | 2011-12-15 | 2011-12-15 | Method for enhancing production intensity of producing itaconic acid by fermentation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011104193368A CN102517343A (en) | 2011-12-15 | 2011-12-15 | Method for enhancing production intensity of producing itaconic acid by fermentation method |
Publications (1)
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|---|---|
| CN102517343A true CN102517343A (en) | 2012-06-27 |
Family
ID=46288409
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|---|---|---|---|
| CN2011104193368A Pending CN102517343A (en) | 2011-12-15 | 2011-12-15 | Method for enhancing production intensity of producing itaconic acid by fermentation method |
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|---|---|
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105274153A (en) * | 2015-11-20 | 2016-01-27 | 江南大学 | Method for increasing yield of itaconic acid produced by fermentation of aspergillus terreus |
-
2011
- 2011-12-15 CN CN2011104193368A patent/CN102517343A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 刘建军等: "发酵法生产衣康酸技术的研究---衣康酸发酵工艺条件的研究(Ⅱ)", 《食品与发酵工业》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105274153A (en) * | 2015-11-20 | 2016-01-27 | 江南大学 | Method for increasing yield of itaconic acid produced by fermentation of aspergillus terreus |
| CN105274153B (en) * | 2015-11-20 | 2020-02-18 | 江南大学 | Method for increasing yield of itaconic acid produced by fermenting aspergillus terreus |
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Application publication date: 20120627 |