CN102864185B - 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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- CN102864185B CN102864185B CN201210347960.6A CN201210347960A CN102864185B CN 102864185 B CN102864185 B CN 102864185B CN 201210347960 A CN201210347960 A CN 201210347960A CN 102864185 B CN102864185 B CN 102864185B
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- citric acid
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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 microorganism fermentation field, specifically, relate to a kind of method of utilizing sorghum flour fermentation production of citric acid.
Background technology
Natural lemon acid distributes very wide at occurring in nature.Plant, all contain citric acid in as the bone of the fruits such as lemon, oranges and tangerines, pineapple and animal, muscle, blood.Within 1784, C.W. house is strangled and first from oranges and tangerines, is extracted citric acid.He is by adding milk of lime to produce citric acid with the method that forms citrate of lime precipitation in fruit is squeezed the juice.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.Within 1913, B. Zha Huosiji report aspergillus niger can generate citric acid.Nineteen twenty-three U.S. Fei Ze company has built the factory that first hand is produced citric acid with aspergillus niger shallow tray fermentation method in the world.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 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 was used that fermentation method produces citric acid the earliest 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 domestic by the main technique route of microorganism fermentation production of citric acid.
Summary of the invention
The object of this invention is to provide a kind of novel method of utilizing sorghum flour fermentation production of citric acid.
In order to realize the object of the invention, a kind of method of utilizing sorghum flour fermentation production of citric acid of the present invention, the microorganism that fermentation adopts is aspergillus niger (Aspergillus niger), the main raw material of fermention medium is liquefaction sorghum flour.
In preceding method, in fermention medium, also added liquefaction Semen Maydis powder, and the weight ratio of liquefaction sorghum flour and Semen Maydis powder is 19-16: 1-4.
In preceding method, in fermention medium, also added nitrogenous source.Described nitrogenous source is inorganic nitrogen-sourced, as ammonium sulfate and/or urea etc.
In preceding method, the sorghum flour in fermention medium and Semen Maydis powder are in advance through liquefaction.During liquefaction, add α-amylase.
In 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's common micro-organisms center, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, 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, ferments 50～65 hours.
In preceding method, the main raw material sorghum flour in fermention medium is that Chinese sorghum crosses 80 mesh sieves after pulverizing.
Object of the present invention can also be further achieved by the following technical measures.
Comprise step:
1) sorghum material pre-treatment: Chinese sorghum, first through pulverizing, crosses 80 mesh sieves after pulverizing.Sorghum flour adds water sizes mixing, and by the powder slurry ratio of seeding tank 15～20%, the powder of fermentor tank 25～35%, starches than preparing.
2) seed culture: the sorghum flour liquefier of the powder slurry ratio containing 15～20% is accessed to aspergillus niger spore as seed culture medium, carry out seed culture, cultivate into aspergillus niger liquid seeds.
3) fermentation culture: cultivate in the fermention medium by the access of cultured aspergillus niger liquid seeds containing the sorghum flour liquefier of 25～35% powder slurry ratio.
It can be aspergillus niger FYCA2358 or other Aspergillus niger strain that citric acid fermentation of the present invention is produced bacterial strain.
Wherein, step 1) in, described powder slurry than refer to the sorghum flour containing in 100 grams of water weight (gram).
Step 2) in, the consisting of of seed culture medium: sorghum flour by 15～20% powder slurry than with 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 to liquefy after (completely liquefaction standard be that iodine liquid is tested constant indigo plant), enter in seeding tank inoculation culture after sterilizing.Seed culture condition is aerlbic culture 24～30 hours at 37 ℃.
Step 3) in, the consisting of of fermention medium: sorghum flour by 25～35% powder slurry than with 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 to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enter fermentor tank and ferment.
In fermention medium, can add appropriate 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.Fermentability reducing sugar is put tank after substantially having consumed.
By the optimization to substratum, find, 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, enter after fermentor tank in the substratum take sorghum flour as main raw material the adaptive phase long, thereby extend 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 increase, shortened fermentation period.In the fermention medium of citric acid, added equally appropriate Semen Maydis powder, its Main Function is to regulate suitable carbon nitrogen source ratio in fermenting process, has improved saccharic acid accretion rate, effectively shortens fermentation period.Through experiment, find, utilize Chinese sorghum to produce citric acid for fermenting raw materials, during fermentation ends, in mash, citric acid acidity can reach 12～15%, and glucose acid invert ratio is greater than 90%.
The inventive method is as producing bacterial strain, to access the fermentative production of carrying out citric acid in the liquid fermentation medium take sorghum flour as main carbon source take aspergillus niger, in fermentor tank, carry out aerlbic culture, 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, especially very applicable in countries such as South America.
Embodiment
Following examples are used for illustrating the present invention, but are not used for limiting the scope of the invention.If do not specialize, the percentage sign " % " relating in embodiment, refers to mass percent; But the per-cent of solution, unless otherwise specified, refers to the grams that contains solute in 100ml solution.
Aspergillus niger (Aspergillus niger) FYCA2358 using in following examples, preserving number is CGMCC No.6465.
Embodiment 1
By in the seed culture medium after citric acid production bacterial classification aspergillus niger FYCA2358 spore access sterilizing.Being formulated as of seed culture medium: sorghum flour (crossing 80 mesh sieves) is prepared by 15% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 15% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enters sterilizing in seeding tank.Seed culture condition is aerlbic culture 28 hours at 37 ℃.After liquid seeds cultivation finishes, the fermention medium after access sterilizing is cultivated.Being formulated as of fermention medium: sorghum flour (crossing 80 mesh sieves) is prepared by 25% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 25% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase to liquefy after (completely liquefaction standard be that iodine liquid is tested constant indigo plant), ferment after entering the sterilizing 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.Fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends, in fermentation liquid, citric acid acidity is 12.5%, glucose acid invert ratio 92%.
Embodiment 2
By in the seed culture medium after citric acid production bacterial classification aspergillus niger FYCA2358 spore access sterilizing.Being formulated as of seed culture medium: sorghum flour (crossing 80 mesh sieves) is prepared by 15% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 15% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 3: 7.Add α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enter sterilizing in seeding tank.Seed culture condition is aerlbic culture 28 hours at 37 ℃.After liquid seeds cultivation finishes, the fermention medium after access sterilizing is cultivated.Being formulated as of fermention medium: sorghum flour (crossing 80 mesh sieves) is prepared by 25% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 25% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase to liquefy after (completely liquefaction standard be that iodine liquid is tested constant indigo plant), ferment after entering the sterilizing 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.Fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends, in fermentation liquid, citric acid acidity is 12.4%, glucose acid invert ratio 92.5%.
Embodiment 3
Citric acid production bacterial classification aspergillus niger FYCA2358 spore is accessed in the seed culture medium after sterilizing under sterile state.Being formulated as of seed culture medium: sorghum flour (crossing 80 mesh sieves) is prepared by 15% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 15% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enters sterilizing in seeding tank.Seed culture condition is aerlbic culture 28 hours at 37 ℃.After liquid seeds cultivation finishes, the fermention medium after access sterilizing is cultivated.Being formulated as of fermention medium: sorghum flour (crossing 80 mesh sieves) is prepared by 25% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 25% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 9: 1.Add α-amylase to liquefy after (completely liquefaction standard be that iodine liquid is tested constant indigo plant), ferment after entering the sterilizing 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.Fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends, in fermentation liquid, citric acid acidity is 14.1%, glucose acid invert ratio 92.5%.
Embodiment 4
Citric acid production bacterial classification aspergillus niger FYCA2358 spore is accessed in the seed culture medium after sterilizing under sterile state.Being formulated as of seed culture medium: sorghum flour (crossing 80 mesh sieves) is prepared by 15% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 15% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enters sterilizing in seeding tank.Seed culture condition is aerlbic culture 28 hours at 37 ℃.After liquid seeds cultivation finishes, the fermention medium after access sterilizing is cultivated.Being formulated as of fermention medium: sorghum flour (crossing 80 mesh sieves) is prepared by 25% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 25% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enter 60m
3after fermentor tank sterilizing, ferment.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.Fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends, in fermentation liquid, citric acid acidity is 14.5%, glucose acid invert ratio 93%.
Embodiment 5
Citric acid production bacterial classification aspergillus niger FYCA2358 spore is inoculated in the seed culture medium after sterilizing under sterile state.Being formulated as of seed culture medium: sorghum flour (crossing 80 mesh sieves) is prepared by 15% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 15% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 9.Add α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enter sterilizing in seeding tank.Seed culture condition is aerlbic culture 28 hours at 37 ℃.After liquid seeds cultivation finishes, the fermention medium after access sterilizing is cultivated.Being formulated as of fermention medium: sorghum flour (crossing 80 mesh sieves) is prepared by 25% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 25% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase to liquefy after (completely liquefaction standard be that iodine liquid is tested constant indigo plant), ferment after entering the sterilizing 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.Fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends, in fermentation liquid, citric acid acidity is 12.5%, glucose acid invert ratio 92%.
Embodiment 6
Citric acid production bacterial classification aspergillus niger FYCA2358 spore is inoculated in the seed culture medium after sterilizing under sterile state.Being formulated as of seed culture medium: sorghum flour (crossing 80 mesh sieves) is prepared by 15% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 15% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 1: 4, adds α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enters sterilizing in seeding tank.Seed culture condition is aerlbic culture 28 hours at 37 ℃.After liquid seeds cultivation finishes, the fermention medium after access sterilizing is cultivated.Being formulated as of fermention medium: sorghum flour (crossing 80 mesh sieves) is prepared by 25% powder slurry ratio tap water, and Semen Maydis powder is starched than preparing with tap water by 25% powder.Wherein the weight ratio of sorghum flour and Semen Maydis powder is 4: 1.Add α-amylase to liquefy after (standard of liquefaction is that iodine liquid is tested constant indigo plant completely), enter 60m
3after fermentor tank sterilizing, ferment.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.Fermentability reducing sugar is put tank after substantially having consumed.
During fermentation ends, in fermentation liquid, citric acid acidity is 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 without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (9)
1. one kind is utilized the method for sorghum flour fermentation production of citric acid, it is characterized in that, the microorganism that fermentation adopts is aspergillus niger (Aspergillus niger) FYCA2358, and its preserving number is CGMCC No.6465, using the main raw material of sorghum flour as fermention medium that liquefy.
2. method according to claim 1, is characterized in that, has also added liquefaction Semen Maydis powder in fermention medium.
3. method according to claim 2, is characterized in that, has also added nitrogenous source in 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 by α-amylase.
7. according to the method described in claim 2-6 any one, it is characterized in that, the weight ratio of liquefy in fermention medium sorghum flour and Semen Maydis powder is 19-16:1-4.
8. according to the method described in claim 1-6 any one, it is characterized in that, 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, ferments 50～65 hours.
9. aspergillus niger (Aspergillus niger) FYCA2358, its preserving number is CGMCC No.6465.
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| CN103695319B (en) * | 2013-12-23 | 2016-02-17 | 安徽丰原发酵技术工程研究有限公司 | A kind ofly produce the bacterial strain of citric acid and the method for preparation of citric acid by fermentation thereof |
| 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 |
| 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 |
| 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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| 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 |
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