CN102864185A - Method for producing citric acid by fermenting sorghum powder - Google Patents
Method for producing citric acid by fermenting sorghum powder Download PDFInfo
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- CN102864185A CN102864185A CN2012103479606A CN201210347960A CN102864185A CN 102864185 A CN102864185 A CN 102864185A CN 2012103479606 A CN2012103479606 A CN 2012103479606A CN 201210347960 A CN201210347960 A CN 201210347960A CN 102864185 A CN102864185 A CN 102864185A
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- citric acid
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- aspergillus niger
- sorghum flour
- semen maydis
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Abstract
The invention provides a method for producing citric acid by fermenting sorghum powder, which comprises the following steps: inoculating the production strain Aspergillus niger into a liquid fermentation culture medium which uses sorghum powder as the main carbon source, and fermenting to produce the citric acid, wherein the culture is carried out while ventilating the fermentation tank; and extracting the fermentation liquid to obtain anhydrous citric acid or citric acid monohydrate crystals. The method for producing citric acid has the advantages of low cost, high yield, stable quality and short production cycle, is an important alternative way for producing citric acid without grains, and is especially suitable for states in South America and the like.
Description
Technical field
The invention belongs to the microbial fermentation field, specifically, relate to a kind of method of utilizing the sorghum flour fermentation production of citric acid.
Background technology
Natural lemon acid distributes very wide at occurring in nature.In the bone of the fruits such as plant such as lemon, oranges and tangerines, pineapple and animal, muscle, blood, all contain citric acid.The C.W. house was strangled and at first extract citric acid from oranges and tangerines in 1784.He produces citric acid by the middle adding milk of lime of squeezing the juice at fruit with the method that forms the citrate of lime precipitation.Fermentation method is produced citric acid and was started from for 19 end of the centurys.C. Wei Gadamer in 1893 finds that mould (genus) bacterium can accumulate citric acid.B. Zha Huosiji report aspergillus niger can generate citric acid in 1913.Nineteen twenty-three U.S. Fei Ze company has built first hand is in the world produced citric acid with aspergillus niger shallow tray fermentation method factory.Belgium, Britain, Germany, the Soviet Union etc. study successful fermentative Production citric acid in succession subsequently.Nineteen fifty-two, this testing laboratory of U.S.'s mayer adopted tank fermentation method scale operation citric acid.After this, tank fermentation method is set up gradually.The submerged fermentation cycle is short, and productive rate is high, saves the labor force, and floor space is little, is convenient to realize instrument control and serialization, has now become the main method of citric acid production.
What China used the earliest that fermentation method produces citric acid is reported as nineteen forty-two Tang Tenghan etc.After 1966, Tianjin Research Institute of Industrial Microbiology, Shanghai City Industry Wei Biological Research Institute are carried out the experimental study of carrying out potato dry powder raw material submerged fermentation citric acid with aspergillus niger in succession, and succeed, thereby determined the domestic main technique route of producing citric acid by microbial fermentation.
Summary of the invention
The purpose of this invention is to provide a kind of novel method of utilizing the sorghum flour fermentation production of citric acid.
In order to realize the object of the invention, a kind of method of utilizing the sorghum flour fermentation production of citric acid of the present invention, the microorganism that fermentation is adopted is aspergillus niger (Aspergillus niger), the main raw material of fermention medium is the liquefaction sorghum flour.
In the preceding method, also added the liquefaction Semen Maydis powder in the fermention medium, and the weight ratio of liquefaction sorghum flour and Semen Maydis powder is 19-16: 1-4.
In the preceding method, also added nitrogenous source in the fermention medium.Described nitrogenous source is inorganic nitrogen-sourced, such as ammonium sulfate and/or urea etc.
In the preceding method, the sorghum flour in the fermention medium and Semen Maydis powder are in advance through liquefaction.Add α-amylase during liquefaction.
In the preceding method, the bacterial strain that fermentation is used is preferably aspergillus niger (Aspergillus niger) FYCA2358.Now be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center, address: No. 3 Institute of Microorganism, Academia Sinica in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode 100101, preservation date on August 21st, 2012, preserving number CGMCC NO.6465.
Fermentation condition is: 35～38 ℃ of temperature, and the volume ratio that passes into air and fermentation liquid is (0.1～0.8): 1, rotating speed 200～400rpm fermented 50～65 hours.
In the preceding method, the main raw material sorghum flour in the fermention medium is that Chinese sorghum crosses 80 mesh sieves through after pulverizing.
Purpose of the present invention can also be further achieved by the following technical measures.
Comprise step:
1) sorghum material pre-treatment: Chinese sorghum crosses 80 mesh sieves first through pulverizing after pulverizing.Sorghum flour adds water sizes mixing, and starches than preparing by the powder slurry ratio of seeding tank 15～20%, the powder of fermentor tank 25～35%.
2) seed culture: the sorghum flour liquefier that will contain 15～20% powder slurry ratio accesses aspergillus niger spore as seed culture medium, carries out seed culture, cultivates into the aspergillus niger liquid seeds.
3) fermentation culture: the access of cultured aspergillus niger liquid seeds contained in the fermention medium of sorghum flour liquefier of 25～35% powder slurry ratio and cultivate.
Citric acid fermentation of the present invention is produced bacterial strain can be aspergillus niger FYCA2358 or other Aspergillus niger strain.
Wherein, step 1) in, described powder slurry is than the weight (gram) of the sorghum flour that refers to contain in 100 gram water.
Step 2) in, the consisting of of seed culture medium: sorghum flour by 15～20% powder slurry than with the tap water preparation, Semen Maydis powder by 15～20% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1-2: 4-3.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enter in the seeding tank inoculation culture after the sterilization.The seed culture condition is 37 ℃ of lower aerlbic cultures 24～30 hours.
Step 3) in, the consisting of of fermention medium: sorghum flour by 25～35% powder slurry than with the tap water preparation, Semen Maydis powder by 25～35% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 19-16: 1-4.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enter fermentor tank and ferment.
In fermention medium, can add an amount of inorganic nitrogen-sourced, such as ammonium sulfate, urea etc.
Described leavening temperature is 35～38 ℃, and the volume ratio that passes into air and fermentation liquid is (0.1～0.8): 1, and mixing speed is 200～400rpm, cultivates pH value nature 50～65 hours.The fermentability reducing sugar is put tank after substantially having consumed.
Find by the optimization to substratum, the seed culture medium of lemon acid-fermentation is take maize powder medium as preferred, if but find that in real attenuation seed culture medium only has Semen Maydis powder, it is long to enter behind the fermentor tank in the substratum take sorghum flour as main raw material the adaptive phase, thereby prolongs fermentation period.Therefore in seed culture medium take Semen Maydis powder as main, add in right amount sorghum flour, bacterium ball fast growth, adaptive faculty increases, and has shortened fermentation period.Added equally an amount of Semen Maydis powder in the fermention medium of citric acid, its Main Function is to regulate carbon nitrogen source ratio suitable in the fermenting process, has improved the saccharic acid accretion rate, effectively shortens fermentation period.Find through experiment, utilize Chinese sorghum to produce citric acid for fermenting raw materials, during fermentation ends in the mash citric acid acidity can reach 12～15%, glucose acid invert ratio is greater than 90%.
The inventive method is as producing the fermentative production of carrying out citric acid in the liquid fermentation medium of bacterial strain access take sorghum flour as main carbon source take aspergillus niger, carry out aerlbic culture in fermentor tank, fermented liquid can obtain Citric Acid, usp, Anhydrous Powder or Citric acid monohydrate Food grade crystallization after extracting.Adopt the inventive method to produce citric acid, cost is low, productive rate is high, steady quality, with short production cycle, is the important alternative route that non-grain is produced citric acid, and is especially very applicable in countries such as South America.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.If do not specialize, the percentage sign that relates among the embodiment " % " refers to mass percent; But the per-cent of solution except as otherwise herein provided, refers to contain in the 100ml solution grams of solute.
The aspergillus niger that uses in following examples (Aspergillus niger) FYCA2358, preserving number is CGMCC No.6465.
Embodiment 1
In the seed culture medium after the citric acid production bacterial classification aspergillus niger FYCA2358 spore access sterilization.Being formulated as of seed culture medium: sorghum flour (cross 80 mesh sieves) by 15% powder slurry than with the tap water preparation, Semen Maydis powder by 15% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase and liquefies after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enters in the seeding tank and sterilizes.The seed culture condition is 37 ℃ of lower aerlbic cultures 28 hours.After liquid seeds was cultivated end, the fermention medium after the access sterilization was cultivated.Being formulated as of fermention medium: sorghum flour (cross 80 mesh sieves) by 25% powder slurry than with the tap water preparation, Semen Maydis powder by 25% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), ferment after entering the sterilization of 500L fermentor tank.In fermention medium, add the inorganic nitrogen-sourced of 0.1% ammonium sulfate.Described leavening temperature is 36 ℃, and the volume ratio that passes into air and fermentation liquid is 0.2: 1, and mixing speed is 400rpm, cultivates pH value nature 65 hours.The fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends in the fermentation liquid citric acid acidity be 12.5%, glucose acid invert ratio 92%.
Embodiment 2
In the seed culture medium after the citric acid production bacterial classification aspergillus niger FYCA2358 spore access sterilization.Being formulated as of seed culture medium: sorghum flour (cross 80 mesh sieves) by 15% powder slurry than with the tap water preparation, Semen Maydis powder by 15% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 3: 7.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enter in the seeding tank and sterilize.The seed culture condition is 37 ℃ of lower aerlbic cultures 28 hours.After liquid seeds was cultivated end, the fermention medium after the access sterilization was cultivated.Being formulated as of fermention medium: sorghum flour (cross 80 mesh sieves) by 25% powder slurry than with the tap water preparation, Semen Maydis powder by 25% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), ferment after entering the sterilization of 500L fermentor tank.In fermention medium, add the inorganic nitrogen-sourced of 0.1% ammonium sulfate.Described leavening temperature is 36 ℃, and the volume ratio that passes into air and fermentation liquid is 0.4: 1, and mixing speed is 400rpm, cultivates pH value nature 67 hours.The fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends in the fermentation liquid citric acid acidity be 12.4%, glucose acid invert ratio 92.5%.
Embodiment 3
With citric acid production bacterial classification aspergillus niger FYCA2358 spore under sterile state, access the sterilization after seed culture medium in.Being formulated as of seed culture medium: sorghum flour (cross 80 mesh sieves) by 15% powder slurry than with the tap water preparation, Semen Maydis powder by 15% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase and liquefies after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enters in the seeding tank and sterilizes.The seed culture condition is 37 ℃ of lower aerlbic cultures 28 hours.After liquid seeds was cultivated end, the fermention medium after the access sterilization was cultivated.Being formulated as of fermention medium: sorghum flour (cross 80 mesh sieves) by 25% powder slurry than with the tap water preparation, Semen Maydis powder by 25% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 9: 1.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), ferment after entering the sterilization of 500L fermentor tank.In fermention medium, add the inorganic nitrogen-sourced of 1% ammonium sulfate.Described leavening temperature is 36 ℃, and the volume ratio that passes into air and fermentation liquid is 0.6: 1, and mixing speed is 400rpm, cultivates pH value nature 61 hours.The fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends in the fermentation liquid citric acid acidity be 14.1%, glucose acid invert ratio 92.5%.
Embodiment 4
With citric acid production bacterial classification aspergillus niger FYCA2358 spore under sterile state, access the sterilization after seed culture medium in.Being formulated as of seed culture medium: sorghum flour (cross 80 mesh sieves) by 15% powder slurry than with the tap water preparation, Semen Maydis powder by 15% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase and liquefies after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enters in the seeding tank and sterilizes.The seed culture condition is 37 ℃ of lower aerlbic cultures 28 hours.After liquid seeds was cultivated end, the fermention medium after the access sterilization was cultivated.Being formulated as of fermention medium: sorghum flour (cross 80 mesh sieves) by 25% powder slurry than with the tap water preparation, Semen Maydis powder by 25% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enter 60m
3Ferment after the fermentor tank sterilization.In fermention medium, add the inorganic nitrogen-sourced of 0.1% ammonium sulfate.Described leavening temperature is 36 ℃, and the volume ratio that passes into air and fermentation liquid is 0.8: 1, and mixing speed is 400rpm, cultivates pH value nature 65 hours.The fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends in the fermentation liquid citric acid acidity be 14.5%, glucose acid invert ratio 93%.
Embodiment 5
With citric acid production bacterial classification aspergillus niger FYCA2358 spore under sterile state, inoculate the sterilization after seed culture medium in.Being formulated as of seed culture medium: sorghum flour (cross 80 mesh sieves) by 15% powder slurry than with the tap water preparation, Semen Maydis powder by 15% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 9.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enter in the seeding tank and sterilize.The seed culture condition is 37 ℃ of lower aerlbic cultures 28 hours.After liquid seeds was cultivated end, the fermention medium after the access sterilization was cultivated.Being formulated as of fermention medium: sorghum flour (cross 80 mesh sieves) by 25% powder slurry than with the tap water preparation, Semen Maydis powder by 25% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), ferment after entering the sterilization of 500L fermentor tank.In fermention medium, add the inorganic nitrogen-sourced of 1% ammonium sulfate.Described leavening temperature is 36 ℃, and the volume ratio that passes into air and fermentation liquid is 0.5: 1, and mixing speed is 300rpm, cultivates pH value nature 69 hours.The fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends in the fermentation liquid citric acid acidity be 12.5%, glucose acid invert ratio 92%.
Embodiment 6
With citric acid production bacterial classification aspergillus niger FYCA2358 spore under sterile state, inoculate the sterilization after seed culture medium in.Being formulated as of seed culture medium: sorghum flour (cross 80 mesh sieves) by 15% powder slurry than with the tap water preparation, Semen Maydis powder by 15% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase and liquefies after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enters in the seeding tank and sterilizes.The seed culture condition is 37 ℃ of lower aerlbic cultures 28 hours.After liquid seeds was cultivated end, the fermention medium after the access sterilization was cultivated.Being formulated as of fermention medium: sorghum flour (cross 80 mesh sieves) by 25% powder slurry than with the tap water preparation, Semen Maydis powder by 25% powder slurry than preparing with tap water.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase and liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant fully), enter 60m
3Ferment after the fermentor tank sterilization.In fermention medium, add the inorganic nitrogen-sourced of 0.1% ammonium sulfate.Described leavening temperature is 36 ℃, and the volume ratio that passes into air and fermentation liquid is 0.6: 1, and mixing speed is 300rpm, cultivates pH value nature 72 hours.The fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends in the fermentation liquid citric acid acidity be 15.1%, glucose acid invert ratio 92.7%.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements all belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. method of utilizing the sorghum flour fermentation production of citric acid, the microorganism that fermentation is adopted is aspergillus niger (Aspergillus niger), it is characterized in that, with the main raw material of liquefaction sorghum flour as fermention medium.
2. method according to claim 1 is characterized in that, has also added the liquefaction Semen Maydis powder in the fermention medium.
3. method according to claim 2 is characterized in that, has also added nitrogenous source in the fermention medium.
4. method according to claim 3 is characterized in that, described nitrogenous source is inorganic nitrogen-sourced.
5. method according to claim 4 is characterized in that, described inorganic nitrogen-sourced be ammonium sulfate and/or urea.
6. method according to claim 2 is characterized in that, liquefies with α-amylase.
7. each described method is characterized in that according to claim 2-6, and the weight ratio of liquefaction sorghum flour and Semen Maydis powder is 19-16: 1-4 in the fermention medium.
8. each described method is characterized in that according to claim 1-6, and fermentation strain is aspergillus niger (Aspergillus niger) FYCA2358, and preserving number is CGMCC No.6465.
9. each described method is characterized in that according to claim 1-6, and fermentation condition is: 35～38 ℃ of temperature, and the volume ratio that passes into air and fermentation liquid is (0.1～0.8): 1, rotating speed 200～400rpm fermented 50～65 hours.
10. aspergillus niger (Aspergillus niger) FYCA2358, its preserving number is CGMCCNo.6465.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103695319A (en) * | 2013-12-23 | 2014-04-02 | 安徽丰原发酵技术工程研究有限公司 | Bacterial strain for producing citric acid and method for preparing citric acid by fermenting same |
| CN105506004A (en) * | 2016-01-05 | 2016-04-20 | 安徽丰原发酵技术工程研究有限公司 | Method for producing citric acid by fermenting konjak powder residues |
| CN105586368A (en) * | 2016-03-16 | 2016-05-18 | 江苏国信协联能源有限公司 | Treatment method of sorghum grains and method for fermentation production of citric acid |
| CN105861575A (en) * | 2016-03-07 | 2016-08-17 | 日照金禾博源生化有限公司 | Citric acid fermentation method |
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| CN102234672A (en) * | 2011-05-09 | 2011-11-09 | 安徽丰原生物化学股份有限公司 | Enzymolysis method for starchy material and method for preparing citric acid |
| CN102373254A (en) * | 2010-08-12 | 2012-03-14 | 中粮生物化学（安徽）股份有限公司 | Enzymolysis method of starchy material and preparation method of citric acid |
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| CN102373254A (en) * | 2010-08-12 | 2012-03-14 | 中粮生物化学（安徽）股份有限公司 | Enzymolysis method of starchy material and preparation method of citric acid |
| CN102234672A (en) * | 2011-05-09 | 2011-11-09 | 安徽丰原生物化学股份有限公司 | Enzymolysis method for starchy material and method for preparing citric acid |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103695319A (en) * | 2013-12-23 | 2014-04-02 | 安徽丰原发酵技术工程研究有限公司 | Bacterial strain for producing citric acid and method for preparing citric acid by fermenting same |
| CN105506004A (en) * | 2016-01-05 | 2016-04-20 | 安徽丰原发酵技术工程研究有限公司 | Method for producing citric acid by fermenting konjak powder residues |
| CN105861575A (en) * | 2016-03-07 | 2016-08-17 | 日照金禾博源生化有限公司 | Citric acid fermentation method |
| CN105861575B (en) * | 2016-03-07 | 2019-09-27 | 日照金禾博源生化有限公司 | A kind of method of citric acid fermentation |
| CN105586368A (en) * | 2016-03-16 | 2016-05-18 | 江苏国信协联能源有限公司 | Treatment method of sorghum grains and method for fermentation production of citric acid |
| CN105586368B (en) * | 2016-03-16 | 2019-05-10 | 江苏国信协联能源有限公司 | A kind of method of the processing method and fermentation production of citric acid of sorghum seed |
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