CN101538603A - Method for producing citric acid by combined fermentation of biological floras - Google Patents
Method for producing citric acid by combined fermentation of biological floras Download PDFInfo
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- CN101538603A CN101538603A CN200910020393A CN200910020393A CN101538603A CN 101538603 A CN101538603 A CN 101538603A CN 200910020393 A CN200910020393 A CN 200910020393A CN 200910020393 A CN200910020393 A CN 200910020393A CN 101538603 A CN101538603 A CN 101538603A
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Abstract
The invention discloses a method for producing citric acid by combined fermentation of biological floras, which is characterized by comprising the following steps: (1) culturing hay bacilli; (2) culturing aspergillus niger; (3) after maize is crashed, mixing the maize with water according to the mass ratio of 1:3, sterilizing the mixture at high temperature in a fermentation cylinder for 30 minutes and cooling the mixture to 28-38 DEG C; and (4) taking cultures prepared in the step (1) and the step (2), together inoculating the cultures in the fermentation cylinder and fermenting the mixture for 60 hours under the conditions that the temperature is between 28 DEG C and 38 DEG C, the ventilation ratio is between 0.1vvm and 0.15vvm and the agitation speed is between 50 rpm and 120 rpm so as to extract the citric acid. The method for producing the citric acid by the combined fermentation of biological floras ensures that the foodstuff utilization rate reaches more than 98 percent and the residual sugar content is below 0.5 percent, saves the foodstuff consumption and improves the productivity of the citric acid and the productivity effect of enterprises.
Description
Technical field
The present invention relates to a kind of production method of citric acid, specifically a kind of method of producing citric acid by combined fermentation of biological floras.
Background technology
Citric acid has another name called Citric Acid, and its molecular formula is C6H8O7, and molecular weight is 192.12, formal name used at school Alpha-hydroxy tricarballylic acid, and water-soluble, ethanol and ether are one of organism main metabolites.It has two kinds of forms: 1, hydrate: melt density 1.542g/cm3 (18 ℃) in the time of 100 ℃; 2, anhydride: clear crystal or powder, 153 ℃ of fusing points have the strong acid flavor.Citric acid is mainly used in foodstuffs industry, and as the acid flavoring and the greasy antioxidant of food, next is used for medicine industry, plastics industry etc.
At present, the citric acid industry is occupied important position in China's organic acid industry, and domestic citric acid production amount accounts for more than 60% of world wide production.Citric acid belongs to the leavened prod of highly energy-consuming, domestic generally is raw material with the corn, through pre-treatment, fermentation production of citric acid, its production technique is: corn → pulverize → size mixing → amylase high temperature liquefaction → fermentation and sterilization → fermentation of Aspergillus niger → citric acid extracts, the deficiency of this production method is: because in process of production, steam instantaneously heating technology is adopted in the liquefaction of amylase high temperature, its destructive force to nutritive ingredient is bigger, cause liquefaction not thorough, residual dextrin is arranged, microorganism can not utilize fully to W-Gum, thereby causes utilization ratio of raw materials below 95%, and residual total reducing sugar is more than 1.5%, cause huge grain consumption and energy dissipation, increased the production cost of enterprise.
Summary of the invention
The object of the present invention is to provide a kind of grain utilization ratio to reach more than 98%, residual sugar content is below 0.5%, saves the grain consumption, can improve the method for producing citric acid by combined fermentation of biological floras of the productive rate of citric acid.
In order to reach above purpose, the technical solution adopted in the present invention is: the method for this a kind of producing citric acid by combined fermentation of biological floras is characterized in that: it may further comprise the steps makes:
(1), the cultivation of Bacillus subtilus (Bacillus subtilis): get and produce the amylase subtilis, be inoculated in the 20% potato solid medium, under 28 ℃ of conditions, cultivated 24 hours, picking list bacterium colony, be inoculated in the 20% potato liquid nutrient medium, shook bottle shaking culture 24 hours under 28 ℃ of conditions, be inoculated in the seeding tank in 1: 10 by volume with the Semen Maydis powder liquid nutrient medium, aeration-agitation is 24 hours under 28 ℃ of conditions;
(2), the cultivation of aspergillus niger (Aspergillus niger): get aspergillus niger, be inoculated in the 20% potato solid medium and cultivated 72 hours, picking list bacterium colony, be inoculated in the wheat bran substratum, under 28 ℃ of conditions, cultivated 10 days,, be inoculated in the Semen Maydis powder liquid nutrient medium seeding tank again with cultured wheat bran bacterial classification, at temperature 28-38 ℃, ventilating ratio is that 0.1vvm and mixing speed are under the condition of 50-120rpm, cultivates 24 hours;
(3), corn pulverized after, press mass ratio 1: 3 and mix with water, high-temperature sterilization is 30 minutes in fermentor tank, is cooled to 35 ℃-38 ℃;
(4), get the culture that step (1) and (2) make, co-inoculation is that 28 ℃-38 ℃, ventilating ratio are that 0.1-0.15vvm and mixing speed are under the condition of 50-120rpm in temperature in fermentor tank, ferments 60 hours, can extract citric acid.
The detection of citric acid content:
The general total acid that detects in the fermenting process, adopt the 0.1429mol/L NaOH solution titration cleaner liquid that fermented, get fermented liquid 1ml, splash into phenolphthalein reagent 2-3 and drip, to the solution becomes light red, the amount of used NaOH promptly is a citric acid content in the fermented liquid with the titration of above-mentioned NaOH solution.
Corn transformation efficiency (%): aspergillus niger generates the gram number of citric acid and the percentage ratio of the ratio of the gram number of consumption sugar.
Consumption sugar: consume the corn quality and take advantage of W-Gum content.
Residual sugar is measured: adopt the fehling's reagent method.
Adopt above-mentioned detection method, the present invention tested:
Shake flask test: get and produce amylase subtilis and aspergillus niger, be inoculated in respectively in the 20% potato solid medium, under 28 degree conditions, cultivated respectively 24 hours and 72 hours, choose single bacterium colony respectively, be inoculated in respectively in 20% potato liquid nutrient medium and the 10% Semen Maydis powder liquid nutrient medium, subtilis was cultivated 24 hours under 28 degree conditions, was inoculated in product citric acid aspergillus niger with 5% inoculum size and shook in the bottle, shake the bottle concussion and cultivated 52 hours, detect citric acid yield and residual sugar liquid measure.According to the shake flask test under the same terms of ten batches, citric acid yield average out to 99.5%, the residual sugar amount is 0.43%.
Common test: get and produce amylase subtilis and aspergillus niger, be inoculated in respectively in the 20% potato solid medium, under 28 degree conditions, cultivated respectively 24 hours and 72 hours, choose single bacterium colony respectively, co-inoculation is in 10% Semen Maydis powder liquid nutrient medium, shake the bottle concussion and cultivated 52 hours, detect citric acid yield and residual sugar liquid measure.According to the shake flask test under the same terms of ten batches, citric acid yield average out to 99.2%, the residual sugar amount is 0.55%.Both coexist this description of test substantially, do not disturb mutually, belong to competitive coexistence.
Learn that through test under conditions suitable, two kinds of bacterial classification symbiotic co-existences utilize mutually, reach the biotic balance state, the corn utilization ratio is more than 98%, and remaining liquid glucose is less than 0.5%.
Beneficial effect of the present invention is: because the present invention does not adopt high temperature liquefaction, the sterilization but the corn after directly will pulverizing is sized mixing, in fermentor tank, utilize and produce the amylase that the amylase subtilis produces, directly Semen Maydis powder is liquefied, the liquid glucose after the liquefaction is produced citric acid for fermentation of Aspergillus niger, and the grain utilization ratio is reached more than 98%, residual sugar content is below 0.5%, saving grain consumes, and has improved the productive rate of citric acid, has improved the productivity effect of enterprise.
Embodiment
Embodiment 1:
The method of this a kind of producing citric acid by combined fermentation of biological floras is characterized in that: it may further comprise the steps makes:
(1), the cultivation of Bacillus subtilus: get and produce the amylase subtilis, be inoculated in the 20% potato solid medium, under 28 ℃ of conditions, cultivated 24 hours, picking list bacterium colony, be inoculated in the 20% potato liquid nutrient medium, shake the bottle concussion and cultivated 24 hours under 28 ℃ of conditions, be inoculated in 1: 10 by volume in 5 cubes of seeding tanks with 20% Semen Maydis powder liquid nutrient medium, aeration-agitation is 24 hours under 28 ℃ of conditions;
(2), the cultivation of aspergillus niger: get aspergillus niger, be inoculated in the 20% potato solid medium and cultivated 72 hours, picking list bacterium colony, be inoculated in the wheat bran substratum, under 28 ℃ of conditions, cultivated 10 days,, be inoculated in 10 cube of 10% Semen Maydis powder liquid nutrient medium seeding tank again with cultured wheat bran bacterial classification, at temperature 28-38 ℃, ventilating ratio is that 0.1vvm and mixing speed are under the condition of 50-120rpm, cultivates 24 hours;
(3), 20 tons of corns are pulverized after, add water and size mixing into 80 cubes, added in 120 cubes of fermentor tanks high-temperature sterilization 30 minutes, be cooled to 35 ℃-38 ℃;
(4), get the culture that step (1) and (2) make, co-inoculation is that 28 ℃-38 ℃, ventilating ratio are that 0.1-0.15vvm and mixing speed are under the condition of 50-120rpm in temperature in 120 cubes of fermentor tanks, ferments 60 hours, can extract citric acid.
Through continuous ten batches fermentative production, the citric acid average yield is more than 99.5%, and the residual sugar amount is reduced to below 0.43%.
Claims (1)
1, a kind of method of producing citric acid by combined fermentation of biological floras is characterized in that: it may further comprise the steps makes:
(1), the cultivation of Bacillus subtilus: get and produce the amylase subtilis, be inoculated in the 20% potato solid medium, under 28 ℃ of conditions, cultivated 24 hours, picking list bacterium colony, be inoculated in the 20% potato liquid nutrient medium, shook bottle shaking culture 24 hours under 28 ℃ of conditions, be inoculated in the seeding tank in 1: 10 by volume with the Semen Maydis powder liquid nutrient medium, aeration-agitation is 24 hours under 28 ℃ of conditions;
(2), the cultivation of aspergillus niger: get aspergillus niger, be inoculated in the 20% potato solid medium and cultivated 72 hours, picking list bacterium colony, be inoculated in the wheat bran substratum, under 28 ℃ of conditions, cultivated 10 days,, be inoculated in the Semen Maydis powder liquid nutrient medium seeding tank again with cultured wheat bran bacterial classification, at temperature 28-38 ℃, ventilating ratio is that 0.1vvm and mixing speed are under the condition of 50-120rpm, cultivates 24 hours;
(3), corn pulverized after, press mass ratio 1: 3 and mix with water, high-temperature sterilization is 30 minutes in fermentor tank, is cooled to 35 ℃-38 ℃;
(4), get the culture that step (1) and (2) make, co-inoculation is that 28 ℃-38 ℃, ventilating ratio are that 0.1-0.15vvm and mixing speed are under the condition of 50-120rpm in temperature in fermentor tank, ferments 60 hours, can extract citric acid.
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| CN200910020393A CN101538603A (en) | 2009-04-29 | 2009-04-29 | Method for producing citric acid by combined fermentation of biological floras |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102250971A (en) * | 2010-12-23 | 2011-11-23 | 日照金禾博源生化有限公司 | Fermentation production method for citric acid |
| CN102352322A (en) * | 2011-11-15 | 2012-02-15 | 南京工业大学 | High-temperature-resistant citric acid producing strain |
| CN102649971A (en) * | 2012-05-15 | 2012-08-29 | 中粮生物化学（安徽）股份有限公司 | Method for cultivating aspergillus niger mouldy bran and method for preparing citric acid through fermentation |
| CN102864182A (en) * | 2012-08-29 | 2013-01-09 | 太仓市茂通化建有限公司 | Method for preparing citric acid by fermenting corn starch by using aspergillus niger |
| CN109415751A (en) * | 2016-05-26 | 2019-03-01 | 密执安大学评议会 | Composition and method for microbial co culture |
-
2009
- 2009-04-29 CN CN200910020393A patent/CN101538603A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102250971A (en) * | 2010-12-23 | 2011-11-23 | 日照金禾博源生化有限公司 | Fermentation production method for citric acid |
| CN102250971B (en) * | 2010-12-23 | 2013-11-27 | 日照金禾博源生化有限公司 | Fermentation production method for citric acid |
| CN102352322A (en) * | 2011-11-15 | 2012-02-15 | 南京工业大学 | High-temperature-resistant citric acid producing strain |
| CN102649971A (en) * | 2012-05-15 | 2012-08-29 | 中粮生物化学（安徽）股份有限公司 | Method for cultivating aspergillus niger mouldy bran and method for preparing citric acid through fermentation |
| CN102864182A (en) * | 2012-08-29 | 2013-01-09 | 太仓市茂通化建有限公司 | Method for preparing citric acid by fermenting corn starch by using aspergillus niger |
| CN109415751A (en) * | 2016-05-26 | 2019-03-01 | 密执安大学评议会 | Composition and method for microbial co culture |
| CN109415751B (en) * | 2016-05-26 | 2023-02-28 | 密执安大学评议会 | Compositions and methods for microbial co-culture |
| US11859230B2 (en) | 2016-05-26 | 2024-01-02 | The Regents Of The University Of Michigan | Compositions and methods for microbial co-culture |
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