CN102250971B - Fermentation production method for citric acid - Google Patents
Fermentation production method for citric acid Download PDFInfo
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- CN102250971B CN102250971B CN2010106025786A CN201010602578A CN102250971B CN 102250971 B CN102250971 B CN 102250971B CN 2010106025786 A CN2010106025786 A CN 2010106025786A CN 201010602578 A CN201010602578 A CN 201010602578A CN 102250971 B CN102250971 B CN 102250971B
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Abstract
The invention discloses a fermentation production method for citric acid, and in particular relates to a continuous fermentation production method for citric acid. A citric acid continuous fermentation system consists of a culture medium storage tank, a seed culture tank, two or more serially connected fermentation tanks and a fermentation mash storage tank. By the citric acid continuous fermentation system, a fermentation period can be saved, the conversion rate of saccharic acid and the utilization rate of the fermentation tanks are improved, and stages such as growth and development and saccharification effects of aspergillus niger on citric acid production bacteria, fermentation and acid production and the like are controllable sectionally.
Description
Technical field
The invention belongs to the production method of citric acid, be specifically related to a kind of production method of continuously fermenting of citric acid.
Background technology
Citric acid is a kind of organic acid of consumption maximum in the world today, and the citric acid of consumption mainly adopts and take the fermentative Production of aspergillus niger as bacterial classification in the world so far.What existing citric acid fermentation technology adopted is batch fermentation pattern intermittently, often completes one batch of fermentation, will blowing and to fermentor tank clean, sterilizing, dnockout and renewed vaccination, just can carry out the next batch fermentation.Cause non-fermentation time to be that the assisted fermentation time takies longer, the fermentor tank utilization ratio is not high, and batch fermentation produces unstablely, and fermentation results fluctuation between batches is larger.
Summary of the invention
For the existing many weak points of existing citric acid batch fermentation technique, the invention provides a kind of production method of continuously fermenting of citric acid.
The production method of continuously fermenting of citric acid of the present invention, its fermenting process is the mode of continuously fermenting; , adopt a plurality of fermentor tank cascade fermentations, at first, to continuous adding bacterial classification and fresh culture in the one grade fermemtation tank, then by the flow velocity identical with charging, one grade fermemtation tank continuous wave output fermentation liquid downwards, step by step, the continuous fermenting process that completes citric acid.
The continuously ferment device of system of citric acid, be comprised of fermentor tank and the fermentation liquid storage tank of substratum storage tank, seed culture tank, a plurality of series connection.The fermentor tank of series connection can be 2, also can be a plurality of, and preferably 4.
The span of control of the technological condition of the continuous ferment process of citric acid of the present invention is as follows:
Leavening temperature: 35-37 ℃;
PH scope: 1.3～4.5;
Material liquid volume in fermentor tank: account for the 75-95% that tank holds;
Ventilation: air quantity 8.5～12m
3/ m
3H.
Other process parameters range can be with reference to existing citric acid fermentation production technology.
Now take and adopt the level Four fermentation mode to be example, illustrating continuously ferments produces concrete technology step and the processing parameter of citric acid:
(1) spawn culture.In seeding tank, add seed culture medium and access aspergillus niger spore, maintain pH3-7, temperature 35-41 ℃, cultivate 12-24 hour, forms the black-koji mould ball stand-by; Air quantity 8.5-10.5m
3/ m
3H.
(2) fermention medium is added in the one grade fermemtation tank, during the 70-80% of fermentation culture fiduciary point fermentor tank volume, start ventilation, stirring and cooling system, make feed temperature in fermentor tank maintain 35-37 ℃, and the inoculum size access of pressing 4-12% weight is from the black-koji mould ball in seeding tank;
(3) by certain flow velocity continuously to the one grade fermemtation tank add fermention medium and by inoculum size 4-12% from the black-koji mould ball in seeding tank, by same flow velocity, flow out or extract fermented liquid out from the one grade fermemtation tank and enter the second order fermentation tank;
(4) when the material liquid volume of second order fermentation tank reaches 75-95% tank appearance, then from the outflow of second order fermentation tank or extraction fermented liquid, enter the three grade fermemtation tank by the flow velocity identical with charging;
(5), when the material liquid volume of three grade fermemtation tank reaches 75-95% tank appearance, from the outflow of three grade fermemtation tank or extraction fermented liquid, enter the level Four fermentor tank with the flow velocity identical with charging;
(6), when the material liquid volume of level Four fermentor tank reaches 75-95% tank appearance, from the outflow of level Four fermentor tank or extraction fermented liquid, enter the fermentation liquid storage tank with the flow velocity identical with charging;
(7) with identical flow velocity, keep continuously the input and output material of cascade fermentation tanks at different levels.
Wherein the described seed culture medium of step (1) is the liquid nutrient medium of carbonaceous sources and nitrogenous source, and total sugar content 10-15% can comprise one or more in cassava, pachyrhizus, liquefied corn, and wheat bran, ammonium sulfate etc.
The described fermention medium of step (2) is the liquid nutrient medium of carbonaceous sources and nitrogenous source, and total sugar content 10-20% can comprise one or more in cassava, pachyrhizus, liquefied corn, and ammonium sulfate etc.
The fermentation parameter of fermentor tanks at different levels is controlled as follows:
The one grade fermemtation tank:
PH2-4.5, temperature 35-37 ℃, the air quantity 8.5-10.5m of sterile air
3/ m
3H.
The second order fermentation tank:
PH1.8-2.9, temperature 35-37 ℃, the air quantity 10.5-12.0m of sterile air
3/ m
3H.
The three grade fermemtation tank:
PH1.5-2.0, temperature 35-37 ℃, the air quantity 9.5-11.0m of sterile air
3/ m
3H.
The level Four fermentor tank:
PH1.3-1.8, temperature 35-37 ℃, the air quantity 8.5-9.5m of sterile air
3/ m
3H.
Because being continuously ferments, and tank volume is the same, so material is the same in the residence time of each tank.
In the situation that it is constant with stirring to control air quantity, the dissolved oxygen in feed liquid changes with fermentation, its rule such as accompanying drawing 1.
In earlier fermentation 10h, growth is slower, and the wear rate of dissolved oxygen is slower, so can control aborning low air quantity, in fermentation 10h～25h, the wear rate of dissolved oxygen comparatively fast needs a large amount of oxygen to supply with growth, so can add aborning Wind Volume.Ferment middle and later stage, oxygen update rate is slow than vegetative period, and dissolved oxygen content maintains the state of rising substantially, so can reduce aborning air quantity.The fermentation air quantity changes over time rather than is unalterable like this, is conducive to save energy.
In fermentor tank due to different stage, the span of control of PH has different, is to adjust respectively according to the Physiology and biochemistry characteristics of aspergillus niger citric acid production bacterium.PH as seeding tank maintains 3-7, be conducive to the spore development growth, the pH of one grade fermemtation tank is conducive to the further growth of black-koji mould ball and produces saccharifying enzyme, make amylolysis become to produce the glucose that bacterium can directly utilize, the pH of second order fermentation tank is conducive to aspergillus niger citric acid production bacterium and enters rapidly the product acid phase, the accumulation citric acid; The pH of feed liquid heterogeneity in same fermentor tank, as above-mentioned one grade fermemtation tank, the upper strata material liquid pH of tank≤4.5, lower floor's material liquid pH >=2, the pH of all the other each tanks is also like this.
The parameters such as the tank pressure of the present invention's fermentor tanks at different levels, ventilation, pH can be stablized regulation and control according to the Physiology and biochemistry characteristics classification of aspergillus niger citric acid production bacterium, and easily are automated.
With batch fermentation, compare, citric acid continuously ferments and has following remarkable advantage: about 8 hours of unproductive time of batch fermentation, comprise cleaning, sterilizing, dnockout and renewed vaccination time, the present invention's mode of continuously fermenting, saved taking of fermentor tank unproductive time, improved the fermentor tank utilization ratio, saved production cost; Owing to being the plural serial stage fermentation, can fully make aspergillus niger citric acid production bacterium utilize the Physiology and biochemistry characteristics of self, shorten fermentation period, improve transformation efficiency.
Batch fermentation and the Data Comparison table that continuously ferments
| Fermentation mode | Fermentation period h | Fermentor tank utilization ratio/% | Glucose acid invert ratio/% | Air quantity is regulated | PH regulator |
| Batch fermentation | 72 | 100 | 101.9 | Generally remain unchanged, air quantity is sometimes required considerably beyond thalline, causes waste | Can't control by the Physiology and biochemistry characteristics of producing bacterium |
| Continuously ferment | 62 | 116 | 104.5 | Required according to thalline, press tank classification regulation and control | According to the Physiology and biochemistry characteristics of producing bacterium, press tank classification regulation and control |
The accompanying drawing explanation:
Dissolved oxygen time diagram when Fig. 1 is 400r/min.
Embodiment
Embodiment 1
At 50m
3In seeding tank, add and contain total reducing sugar 12.5%(m/m) seed culture medium and access aspergillus niger spore, maintain pH3-7,40 ℃ of temperature, cultivate 12-24 hour, forms the black-koji mould ball stand-by.
To contain total reducing sugar 12.5%(m/m) the corn fermentation substratum add 250m
3In the one grade fermemtation tank that tank holds, fermentation culture fiduciary point fermentor tank volume 70% the time, start ventilation, stirring and cooling system, make feed temperature in fermentor tank maintain 35-37 ℃ and by 10% inoculum size access from the black-koji mould ball in seeding tank, maintain air quantity 8.6 m
3/ m
3H, pH2.9-4.5;
Press 13.5m
3The flow velocity of/h continuously to the one grade fermemtation tank add fermention medium and in 10% ratio from the black-koji mould ball in seeding tank, press 14.85m
3The flow velocity of/h flows out or extracts fermented liquid out from the one grade fermemtation tank and enters 250m
3The second order fermentation tank that tank holds, maintain air quantity 11.5 m
3/ m
3H, temperature 35-37 ℃, pH2.0-2.9;
When the material liquid volume of second order fermentation tank reaches 80% tank appearance, then press 14.85m
3The flow velocity of/h flows out or extracts fermented liquid out from the second order fermentation tank and enters 250m
3The three grade fermemtation tank that tank holds, maintain air quantity 11.0 m
3/ m
3H, temperature 35-37 ℃, pH1.5-2.0;
When the material liquid volume of three grade fermemtation tank reaches 80% tank appearance, from the outflow of three grade fermemtation tank or extraction fermented liquid, enter 250m with the flow velocity identical with charging
3The level Four fermentor tank that tank holds, maintain air quantity 9.0 m
3/ m
3H, temperature 35-37 ℃, pH1.3-1.5;
When the material liquid volume of level Four fermentor tank reaches 80% tank appearance, from the outflow of level Four fermentor tank or extraction fermented liquid, enter 100m with the flow velocity identical with charging
3The fermentation liquid storage tank;
With identical flow velocity, keep continuously the input and output material of cascade fermentation tanks at different levels, after continuous operation 240 hours, surveying fermentation liquid Citric acid monohydrate Food grade concentration in the fermentation liquid storage tank is 13.02g/100g, glucose acid invert ratio is 104.2%, and be 12.73g/100g with the fermentor tank batch fermentation monthly average Citric acid monohydrate Food grade concentration of volume, average glucose acid invert ratio is 101.9%.
Embodiment 2
At 50m
3In seeding tank, add and contain total reducing sugar 13.0%(m/m) seed culture medium and access aspergillus niger spore, maintain pH3-7,38 ℃ of temperature, cultivate 12-24 hour, forms the black-koji mould ball stand-by.
To contain total reducing sugar 13.5%(m/m) the corn fermentation substratum add 250m
3In the one grade fermemtation tank that tank holds, fermentation culture fiduciary point fermentor tank volume 80% the time, start ventilation, stirring and cooling system, make feed temperature in fermentor tank maintain 35-37 ℃ and by 8% inoculum size access from the black-koji mould ball in seeding tank, maintain air quantity 9.8 m
3/ m
3H, pH2.9-4.5;
Press 13m
3The flow velocity of/h continuously to the one grade fermemtation tank add fermention medium and in 8% ratio from the black-koji mould ball in seeding tank, 14.85m
3The flow velocity of/h flows out or extracts fermented liquid out from the one grade fermemtation tank and enters 250m
3The second order fermentation tank that tank holds, maintain air quantity 12.0 m
3/ m
3H, temperature 35-37 ℃, pH2.0-2.9;
When the material liquid volume of second order fermentation tank reaches 88% tank appearance, then 14.85m
3The flow velocity of/h flows out or extracts fermented liquid out from the second order fermentation tank and enters 250m
3The three grade fermemtation tank that tank holds, maintain air quantity 11.2 m
3/ m
3H, temperature 35-37 ℃, pH1.5-2.0;
When the material liquid volume of three grade fermemtation tank reaches 88% tank appearance, from the outflow of three grade fermemtation tank or extraction fermented liquid, enter 250m with the flow velocity identical with charging
3The level Four fermentor tank that tank holds, maintain air quantity 8.5 m
3/ m
3H, temperature 35-37 ℃, pH1.3-1.5;
When the material liquid volume of level Four fermentor tank reaches 88% tank appearance, from the outflow of level Four fermentor tank or extraction fermented liquid, enter 100m with the flow velocity identical with charging
3The fermentation liquid storage tank;
With identical flow velocity, keep continuously the input and output material of cascade fermentation tanks at different levels, after continuous operation 240 hours, surveying fermentation liquid Citric acid monohydrate Food grade concentration in the fermentation liquid storage tank is 13.45g/100g, glucose acid invert ratio is 103.5%, and be 12.73g/100g with the fermentor tank batch fermentation monthly average Citric acid monohydrate Food grade concentration of volume, average glucose acid invert ratio is 101.9%.
Embodiment 3
At 75m
3In seeding tank, add and contain total reducing sugar 12.5%(m/m) seed culture medium and access aspergillus niger spore, maintain pH3-7,40 ℃ of temperature, cultivate 12-24 hour, forms the black-koji mould ball stand-by.
To contain total reducing sugar 12.5%(m/m) the cassava fermention medium add 550m
3In the one grade fermemtation tank that tank holds, fermentation culture fiduciary point fermentor tank volume 75% the time, start ventilation, stirring and cooling system, make feed temperature in fermentor tank maintain 35-37 ℃ and by 10% inoculum size access from the black-koji mould ball in seeding tank, maintain air quantity 8.5 m
3/ m
3H, pH2.9-4.5;
Press 31m
3The flow velocity of/h continuously to the one grade fermemtation tank add fermention medium and in 10% ratio from the black-koji mould ball in seeding tank, press 34.1m
3The flow velocity of/h flows out or extracts fermented liquid out from the one grade fermemtation tank and enters 550m
3The second order fermentation tank that tank holds, maintain air quantity 10.8 m
3/ m
3H, temperature 35-37 ℃, pH2.0-2.9;
When the material liquid volume of second order fermentation tank reaches 85% tank appearance, then press 34.1m
3The flow velocity of/h flows out or extracts fermented liquid out from the second order fermentation tank and enters 550m
3The three grade fermemtation tank that tank holds, maintain air quantity 10.0 m
3/ m
3H, temperature 35-37 ℃, pH1.5-2.0;
When the material liquid volume of three grade fermemtation tank reaches 85% tank appearance, from the outflow of three grade fermemtation tank or extraction fermented liquid, enter 550m with the flow velocity identical with charging
3The level Four fermentor tank that tank holds, maintain air quantity 9.0 m
3/ m
3H, temperature 35-37 ℃, pH1.3-1.5;
When the material liquid volume of level Four fermentor tank reaches 85% tank appearance, from the outflow of level Four fermentor tank or extraction fermented liquid, enter 100m with the flow velocity identical with charging
3The fermentation liquid storage tank;
With identical flow velocity, keep continuously the input and output material of cascade fermentation tanks at different levels, after continuous operation 240 hours, surveying fermentation liquid Citric acid monohydrate Food grade concentration in the fermentation liquid storage tank is 13.06g/100g, glucose acid invert ratio is 104.5%, and be 12.73g/100g with the fermentor tank batch fermentation monthly average Citric acid monohydrate Food grade concentration of volume, average glucose acid invert ratio is 101.9%.
Claims (1)
1. the fermentation method for producing of a citric acid, is characterized in that fermenting process is the mode of continuously fermenting; Adopt a plurality of fermentor tank cascade fermentations, at first, to continuous adding bacterial classification and fresh culture in the one grade fermemtation tank, then by the flow velocity identical with charging, one grade fermemtation tank continuous wave output fermentation liquid downwards, step by step, the continuous fermenting process that completes citric acid; The span of control of technological condition is as follows: leavening temperature: 35～37 ℃; PH scope: 1.3～4.5; Material liquid volume in fermentor tank: account for 75～95% of tank appearance; Ventilation: air quantity 8.5～12m
3/ m
3H;
Concrete technology step and processing parameter:
(1) spawn culture: in seeding tank, add seed culture medium and access aspergillus niger spore, maintain pH3-7, temperature 35-41 ℃, cultivate 12-24 hour, forms the black-koji mould ball stand-by; Air quantity 8.5-10.5m
3/ m
3H;
(2) fermention medium is added in the one grade fermemtation tank, during the 70-80% of fermentation culture fiduciary point fermentor tank volume, start ventilation, stirring and cooling system, make feed temperature in fermentor tank maintain 35-37 ℃, and the inoculum size access of pressing 4-12% weight is from the black-koji mould ball in seeding tank;
(3) by certain flow velocity continuously to the one grade fermemtation tank add fermention medium and by inoculum size 4-12% from the black-koji mould ball in seeding tank, by same flow velocity, flow out or extract fermented liquid out from the one grade fermemtation tank and enter the second order fermentation tank;
(4) when the material liquid volume of second order fermentation tank reaches 75-95% tank appearance, then from the outflow of second order fermentation tank or extraction fermented liquid, enter the three grade fermemtation tank by the flow velocity identical with charging;
(5), when the material liquid volume of three grade fermemtation tank reaches 75-95% tank appearance, from the outflow of three grade fermemtation tank or extraction fermented liquid, enter the level Four fermentor tank with the flow velocity identical with charging;
(6), when the material liquid volume of level Four fermentor tank reaches 75-95% tank appearance, from the outflow of level Four fermentor tank or extraction fermented liquid, enter the fermentation liquid storage tank with the flow velocity identical with charging;
(7) with identical flow velocity, keep continuously the input and output material of cascade fermentation tanks at different levels;
The fermentation parameter of one grade fermemtation tank is controlled as follows: pH2-4.5, temperature 35-37 ℃, the air quantity 8.5-10.5m of sterile air
3/ m
3H;
The fermentation parameter of second order fermentation tank is controlled as follows: pH1.8-2.9, temperature 35-37 ℃, the air quantity 10.5-12.0m of sterile air
3/ m
3H;
The fermentation parameter of three grade fermemtation tank is controlled as follows: pH1.5-2.0, temperature 35-37 ℃, the air quantity 9.5-11.0m of sterile air
3/ m
3H;
The fermentation parameter of level Four fermentor tank is controlled as follows: pH1.3-1.8, temperature 35-37 ℃, the air quantity 8.5-9.5m of sterile air
3/ m
3H.
2. the fermentation method for producing of citric acid as claimed in claim 1, it is characterized in that described seed culture medium is the liquid nutrient medium of carbonaceous sources and nitrogenous source, total sugar content 10-15%, comprise one or more in cassava, pachyrhizus, liquefied corn, and wheat bran, ammonium sulfate.
3. the fermentation method for producing of citric acid as claimed in claim 1, is characterized in that described fermention medium is the liquid nutrient medium of carbonaceous sources and nitrogenous source, and total sugar content 10-20% comprises one or more in cassava, pachyrhizus, liquefied corn and ammonium sulfate.
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| CN102250971B true CN102250971B (en) | 2013-11-27 |
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104277977B (en) * | 2013-07-01 | 2017-06-30 | 中粮生物化学（安徽）股份有限公司 | A kind of preparation method of aspergillus niger spore suspension and the storage method of aspergillus niger spore |
| CN104531541B (en) * | 2014-12-16 | 2017-12-22 | 日照金禾博源生化有限公司 | A kind of citric acid fermentation seed tank culture technique |
| CN105296549A (en) * | 2015-10-23 | 2016-02-03 | 安徽丰原发酵技术工程研究有限公司 | Method for improving citric acid fermenting level |
| CN107868803B (en) * | 2017-10-23 | 2021-03-23 | 日照金禾博源生化有限公司 | Method for reusing citric acid fermentation mycelium hydrolysate for citric acid fermentation |
| CN107815475A (en) * | 2017-12-08 | 2018-03-20 | 江苏国信协联能源有限公司 | A kind of method by two benches fermentation production of citric acid zymotic fluid |
| CN109055444B (en) * | 2018-08-28 | 2020-06-05 | 江苏国信协联能源有限公司 | Aspergillus niger seed continuous culture and citric acid production method |
| CN112481315B (en) * | 2019-09-11 | 2023-05-12 | 吉林中粮生化有限公司 | Method for preparing citric acid by continuous liquid-changing fermentation |
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