CN101638675B - Method for manufacturing citric acid by cane sugar fermentation method - Google Patents
Method for manufacturing citric acid by cane sugar fermentation method Download PDFInfo
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Abstract
The invention provides a method for manufacturing citric acid by a cane fermentation method. In the method of the invention, the cane sugar is utilized as the carbon source, and inorganic and organic nitrogen sources and various nutrient elements are added for fermentation to manufacture the citric acid. The quality of the fermentation liquid is obviously improved; the metabolic rate of the saccharic acid is improved; the fermentation cycle is shortened; the production cost is minimized; the acidity of citric acid in the fermentation liquid can reach 11-14%; the fermentation cycle is 70-85 hours; the conversion percent of the saccharic acid is greater than 89%. The invention has positive meanings to promote the diversification of the production raw materials of the citric acid and in particular the development of foreign markets.
Description
Technical field
The present invention relates to a kind of working method of Hydrocerol A, specifically, relate to a kind of method of utilizing sucrose to carry out fermentation production of citric acid.
Background technology
Hydrocerol A and sodium salt etc. thereof are one of pillar products of fermentation industry, in recent years, China's citric acid production scale develop rapidly, the YO of Hydrocerol A and salt thereof has reached 450,000 tons, is the maximum product of output in the organic acid leavened prod, wherein 80% exports to foreign countries.China's citric acid production raw material mainly with grain as the master, China has a large population and a few land, crisis in food is serious day by day.The raw material long run supply of Hydrocerol A is not enough, and corn is under-supply when special, and the market value of Hydrocerol A will rise always.
In recent years, sucrose yield is the trend of rising progressively year by year, and especially the sucrose price on the foreign market is to be lower than the Dian Fentang price of China.Therefore, carry out the citric acid fermented Study on Technology of sucrose, guarantee that grain-production safety has crucial meaning for alleviating the pressure of domestic citric acid production with grain.
Summary of the invention
The purpose of this invention is to provide a kind of method of utilizing sucrose to carry out fermentation production of citric acid, this method utilizes sucrose to be raw material, has improved the saccharic acid metabolic rate, has shortened fermentation period, and the fermented liquid quality is able to obvious raising.
In order to realize the object of the invention, the method for a kind of manufacturing citric acid by cane sugar fermentation method of the present invention, it adopts and earlier cultured black mold wheat bran spore is carried out seed culture in the seed culture medium that contains 7～12% sucrose, cultivates into the black mold liquid seeds; And then in the fermention medium that contains 14～16% sucrose, cultivate.
Specifically, it comprises the steps:
1) flat board or slant culture: citric acid production bacterial classification black mold is inoculated into wort agar flat board or slant medium cultivation;
2) wheat bran is cultivated: the Hydrocerol A bacterial classification black mold after will cultivating then inserts a ring aspergillus niger spore, in the wheat bran seed culture medium, cultivates, and cultivates into the wheat bran spore;
3) seed culture: the seed culture medium that will contain 7～12% concentration expressed in percentage by weight sucrose inserts the wheat bran spore, carries out seed culture, cultivates into the black mold liquid seeds;
4) fermentation culture: cultured black mold liquid seeds inserted in the fermention medium that contains 14～16% concentration expressed in percentage by weight sucrose cultivate.
Wherein, citric acid production bacterial classification according to the invention is black mold Co827.
In the step 1), the concentration of said wort is 2～4 mother-in-law U.S. degree.
Step 2) in, said wheat bran seed culture medium consists of: wheat bran 20～30%, sal epsom 0.006～0.02%, potassium primary phosphate 0.01～0.03%, pH nature, tap water preparation.Percentage ratio (%) is weight percentage, and is as follows.
The wheat bran culture condition is 37 ℃ and cultivated 7～10 days down.
In the step 3), the consisting of of seed culture medium: sucrose 7～12%, steeping water 0.4～0.9%, potassium primary phosphate 0.01～0.05%, ammonium sulfate 0.006～0.02%, sal epsom 0.006～0.02%, pH nature, tap water preparation.
The seed culture condition is 37 ℃ of cultivations 24～30 hours of ventilating down.
In the step 4), sucrose 14～16%, steeping water 0.5～1.0%, potassium primary phosphate 0.005～0.01%, ammonium sulfate 0.01～0.02%, sal epsom 0.01～0.02%, pH nature, tap water preparation.
The required carbon source of fermentation culture according to the invention is: sucrose; Nitrogenous source is: ammonium sulfate, steeping water.
Said leavening temperature is 35～39 ℃, and pH is 2.5～6.0, and ventilating ratio is 1: (0.1～0.6), mixing speed are 300～500rpm, cultivate 70～85 hours, put jar when the fermentability reducing sugar has consumed basically.
The Hydrocerol A acidity that the present invention utilizes the sucrose fermentation method to obtain reaches 11～14%, and glucose acid invert ratio is greater than 89%.
The present invention utilizes the method for manufacturing citric acid by cane sugar fermentation method, has following advantage:
1. the present invention optimizes SM;
2. in the SM; Nitrogenous source except that carbon source and nutritive element are relatively poorer; In experimentation, be utilized in the line traffic control means; Culture media nitrogen source and nutritive element to citric acid fermentation are optimized, and make cost minimization, and set up Hydrocerol A bacterial classification growth metabolism process and fermenting process modeling and control techniques;
3. the present invention utilizes sucrose for carbon source and add inorganic and organic nitrogen source, various nutritive element, at growth, the metabolic different steps demand difference to oxygen, various nutritive element, various carbon source, nitrogenous source, carries out fermentation production of citric acid according to mikrobe; The fermented liquid quality obviously improves; Improved the saccharic acid metabolic rate, shortened fermentation period, Hydrocerol A acidity reaches 11～14% in the fermented liquid; Fermentation period 70～85 hours, glucose acid invert ratio is greater than 89%; To opening up the variation of citric acid production raw material, especially the exploitation of foreign market has positive effect.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.
Embodiment 1
Earlier citric acid production bacterial classification black mold Co827 spore inoculating is cultivated to wort (concentration is 4 mother-in-law U.S. degree) agar plate; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating under sterile state, to insert a ring aspergillus niger spore to containing wheat bran 30%, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.01%, was cultivated 9 days for 37 ℃.To cultivate sophisticated wheat bran spore and be linked into and contain sucrose 10.5%, steeping water 0.6%, potassium primary phosphate 0.03%, ammonium sulfate 0.01%, in the liquid seed culture medium of sal epsom 0.008%, pH nature, tap water preparation.37 ℃, ventilate and cultivated 28 hours.Treat liquid seeds cultivate good after, inoculate and contain sucrose 14%, steeping water 0.5%, potassium primary phosphate 0.03%; Ammonium sulfate 0.01% is cultivated in the 5L ferment tank substratum of sal epsom 0.015%, and temperature is 37 ℃, and pH is 4.0; Sterile air is that 1: 0.4 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 350rpm, ventilates to cultivating 80 hours; Residual reducing sugar 1.05%, Hydrocerol A acidity reaches 11.9%, glucose acid invert ratio 89%.
Embodiment 2
Earlier citric acid production bacterial classification black mold Co827 spore inoculating is cultivated to wort (concentration is 4 mother-in-law U.S. degree) agar plate; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating under sterile state, to insert a ring aspergillus niger spore to containing wheat bran 20%, sal epsom 0.006% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.01%, was cultivated 10 days for 37 ℃.To cultivate sophisticated wheat bran spore and be linked into and contain sucrose 10.5%, steeping water 0.6%, potassium primary phosphate 0.03%, ammonium sulfate 0.01%, in the liquid seed culture medium of sal epsom 0.01%, pH nature, tap water preparation.37 ℃, ventilate and cultivated 26 hours.Treat liquid seeds cultivate good after, inoculate and contain sucrose 14.5%, steeping water 0.6%, potassium primary phosphate 0.05%; Ammonium sulfate 0.01% is cultivated in the 5L ferment tank substratum of sal epsom 0.02%, and temperature is 37 ℃, and pH is 3.0; Sterile air is that 1: 0.4 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 400rpm, ventilates to cultivating 77 hours; Residual reducing sugar 0.98%, Hydrocerol A acidity reaches 11.89%, glucose acid invert ratio 89.2%.
Embodiment 3
Earlier citric acid production bacterial classification black mold Co827 spore inoculating is cultivated to wort (concentration is 4 mother-in-law U.S. degree) agar plate; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating under sterile state, to insert a ring aspergillus niger spore to containing wheat bran 30%, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.02%, was cultivated 7 days for 37 ℃.To cultivate sophisticated wheat bran spore and be linked into and contain sucrose 9%, steeping water 0.8%, potassium primary phosphate 0.01%, ammonium sulfate 0.02%, in the liquid seed culture medium of sal epsom 0.009%, pH nature, tap water preparation.37 ℃, ventilate and cultivated 29 hours.Treat liquid seeds cultivate good after, inoculate and contain sucrose 15%, steeping water 0.5%, potassium primary phosphate 0.01%; Ammonium sulfate 0.01% is cultivated in the 5L ferment tank substratum of sal epsom 0.015%, and temperature is 37 ℃, and pH is 4.0; Sterile air is that 1: 0.1 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 300rpm, ventilates to cultivating 82 hours; Residual reducing sugar 0.95%, Hydrocerol A acidity reaches 12.8%, glucose acid invert ratio 89.4%.
Embodiment 4
Earlier citric acid production bacterial classification black mold Co827 spore inoculating is cultivated to wort (concentration is 2 mother-in-law U.S. degree) agar plate; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating under sterile state, to insert a ring aspergillus niger spore to containing wheat bran 25%, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.01%, was cultivated 9 days for 37 ℃.To cultivate sophisticated wheat bran spore and be linked into and contain sucrose 12%, steeping water 0.4%, potassium primary phosphate 0.05%, ammonium sulfate 0.01%, in the liquid seed culture medium of sal epsom 0.01%, pH nature, tap water preparation.37 ℃, ventilate and cultivated 25 hours.Treat liquid seeds cultivate good after, inoculate and contain sucrose 14.75%, steeping water 1.0%, potassium primary phosphate 0.05%; Ammonium sulfate 0.02% is cultivated in the 5L ferment tank substratum of sal epsom 0.02%, and temperature is 38.5 ℃, and pH is 6.0; Sterile air is that 1: 0.6 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 450rpm, ventilates to cultivating 78 hours; Residual reducing sugar 1.15%, Hydrocerol A acidity reaches 13.59%, glucose acid invert ratio 89.65%.
Embodiment 5
Earlier citric acid production bacterial classification black mold Co827 spore inoculating is cultivated to wort (concentration is 4 mother-in-law U.S. degree) agar plate; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating under sterile state, to insert a ring aspergillus niger spore to containing wheat bran 30%, sal epsom 0.01% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.03%, was cultivated 9 days for 37 ℃.To cultivate sophisticated wheat bran spore and be linked into and contain sucrose 8%, steeping water 0.9%, potassium primary phosphate 0.02%, ammonium sulfate 0.02%, in the liquid seed culture medium of sal epsom 0.012%, pH nature, tap water preparation.37 ℃, ventilate and cultivated 30 hours.Treat liquid seeds cultivate good after, inoculate and contain sucrose 16%, steeping water 0.5%, potassium primary phosphate 0.01%; Ammonium sulfate 0.02% is cultivated in the 50L ferment tank substratum of sal epsom 0.02%, and temperature is 37 ℃, and pH is 2.5; Sterile air is that 1: 0.3 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 400rpm, ventilates to cultivating 75 hours; Residual reducing sugar 0.89%, Hydrocerol A acidity reaches 13.36%, glucose acid invert ratio 89.23%.
Embodiment 6
Earlier citric acid production bacterial classification black mold Co827 spore inoculating is cultivated to wort (concentration is 4 mother-in-law U.S. degree) agar plate; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating under sterile state, to insert a ring aspergillus niger spore to containing wheat bran 30%, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.01%, was cultivated 9 days for 37 ℃.To cultivate sophisticated wheat bran spore and be linked into and contain sucrose 7%, steeping water 0.9%, potassium primary phosphate 0.03%, ammonium sulfate 0.02%, in the liquid seed culture medium of sal epsom 0.008%, pH nature, tap water preparation.37 ℃, ventilate and cultivated 28 hours.Treat liquid seeds cultivate good after, inoculate and contain sucrose 14.8%, steeping water 0.8%, potassium primary phosphate 0.01%; Ammonium sulfate 0.01% is cultivated in the 50L ferment tank substratum of sal epsom 0.02%, and temperature is 39 ℃, and pH is 4.5; Sterile air is that 1: 0.4 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 400rpm, ventilates to cultivating 76 hours; Residual reducing sugar 1.05%, Hydrocerol A acidity reaches 13.9%, glucose acid invert ratio 89.6%.
Embodiment 7
Earlier citric acid production bacterial classification black mold Co827 spore inoculating is cultivated to wort (concentration is 4 mother-in-law U.S. degree) agar plate; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating under sterile state, to insert a ring aspergillus niger spore to containing wheat bran 30%, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.01%, was cultivated 9 days for 37 ℃.To cultivate sophisticated wheat bran spore and be linked into and contain sucrose 10.5%, steeping water 0.6%, potassium primary phosphate 0.03%, ammonium sulfate 0.01%, in the liquid seed culture medium of sal epsom 0.02%, pH nature, tap water preparation.37 ℃, ventilate and cultivated 28 hours.Treat liquid seeds cultivate good after, inoculate and contain sucrose 15%, steeping water 0.5%, potassium primary phosphate 0.008%; Ammonium sulfate 0.01% is cultivated in the 500L ferment tank substratum of sal epsom 0.02%, and temperature is 37 ℃, and pH is 6.0; Sterile air is that 1: 0.3 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 350rpm, ventilates to cultivating 73 hours; Residual reducing sugar 1.07%, Hydrocerol A acidity reaches 14.0%, glucose acid invert ratio 89.7%.
Though, the present invention has been done detailed description in the preceding text with general explanation and specific embodiments, on basis of the present invention, can to some modifications of do or improvement, this will be apparent to those skilled in the art.Therefore, these modifications or the improvement on the basis of not departing from spirit of the present invention, made all belong to the scope that requirement of the present invention is protected.
Claims (1)
1. the method for a manufacturing citric acid by cane sugar fermentation method; It is characterized in that; It adopts and earlier cultured black mold Co827 wheat bran spore is carried out seed culture in the seed culture medium that contains 7～12% concentration expressed in percentage by weight sucrose, cultivates into black mold Co827 liquid seeds; And then in the fermention medium that contains 14～16% concentration expressed in percentage by weight sucrose, cultivate;
It comprises the steps:
1) flat board or slant culture: citric acid production bacterial classification black mold is inoculated into wort agar flat board or slant medium cultivation;
2) wheat bran is cultivated: the Hydrocerol A bacterial classification black mold after will cultivating then inserts a ring aspergillus niger spore, in the wheat bran seed culture medium, cultivates, and cultivates into the wheat bran spore;
3) seed culture: the seed culture medium that will contain 7～12% concentration expressed in percentage by weight sucrose inserts the wheat bran spore, carries out seed culture, cultivates into the black mold liquid seeds;
4) fermentation culture: cultured black mold liquid seeds inserted in the fermention medium that contains 14～16% concentration expressed in percentage by weight sucrose cultivate;
Step 2) in, said wheat bran seed culture medium consists of: wheat bran 20～30%, and sal epsom 0.006～0.02%, potassium primary phosphate 0.01～0.03%, the pH nature, the tap water preparation, said wheat bran culture condition is 37 ℃ and cultivated 7～10 days down;
In the step 3), the consisting of of seed culture medium: sucrose 7～12%, steeping water 0.4～0.9%; Potassium primary phosphate 0.01～0.05%; Ammonium sulfate 0.006～0.02%, sal epsom 0.006～0.02%, pH nature; Tap water preparation, said seed culture condition are 37 ℃ of cultivations 24～30 hours of ventilating down;
The consisting of of fermention medium in the step 4): sucrose 14～16%, steeping water 0.5～1.0%, potassium primary phosphate 0.005～0.01%; Ammonium sulfate 0.01～0.02%, sal epsom 0.01～0.02%, pH nature; The tap water preparation, leavening temperature described in the step 4) is 35～39 ℃, pH is 2.5～6.0; Ventilating ratio is 1: (0.1～0.6), mixing speed are 300～500rpm, cultivate 70～85 hours.
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| CN102191282B (en) * | 2011-03-29 | 2014-07-23 | 安徽丰原发酵技术工程研究有限公司 | Method for producing malic acid by fermentation of citric acid mother liquor |
| CN102242160B (en) * | 2011-05-11 | 2013-07-10 | 安徽丰原发酵技术工程研究有限公司 | Method for fermenting and producing L-malic acid with raw material of citric acid broth |
| CN103074235A (en) * | 2012-12-28 | 2013-05-01 | 安徽丰原发酵技术工程研究有限公司 | Method for screening high-yield citric acid strains by saccharifying enzyme |
| CN103667372A (en) * | 2013-11-19 | 2014-03-26 | 天津市工业微生物研究所 | Method for preparing bran starter for citric acid by liquid inoculation |
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| CN1393564A (en) * | 2001-07-04 | 2003-01-29 | 上海新立工业微生物科技有限公司 | Process for preparing citric acd by fermentation and bacterial strain for it |
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| CN1393564A (en) * | 2001-07-04 | 2003-01-29 | 上海新立工业微生物科技有限公司 | Process for preparing citric acd by fermentation and bacterial strain for it |
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