CN101638674B - Method for manufacturing citric acid by utilizing cane sugar hydrolysate fermentation method - Google Patents
Method for manufacturing citric acid by utilizing cane sugar hydrolysate fermentation method Download PDFInfo
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- CN101638674B CN101638674B CN2009100916144A CN200910091614A CN101638674B CN 101638674 B CN101638674 B CN 101638674B CN 2009100916144 A CN2009100916144 A CN 2009100916144A CN 200910091614 A CN200910091614 A CN 200910091614A CN 101638674 B CN101638674 B CN 101638674B
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Abstract
The invention provides a method for manufacturing citric acid by utilizing cane sugar hydrolysate fermentation method. In the method, glucose in the cane sugar hydrolysate and a small amount of cane sugar and fructose are utilized as carbon source, and inorganic and organic nitrogen source and various nutrient elements are added for fermentation to manufacture the citric acid. In the method of the invention, the nitrogen source of fermentation production culture medium is optimized to ensure that the production cost is minimized. The cane sugar hydrolysate is utilized as raw materials in the invention and the quality of the fermentation liquid is obviously improved, thus having 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 production method of citric acid, specifically, relate to a kind of method of utilizing cane sugar hydrolysate to carry out fermentation production of citric acid.
Background technology
Citric acid 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 annual production of citric acid and salt thereof has reached 450,000 tons, is the product of output maximum 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 deficiency of citric acid, corn is under-supply when special, and the market value of citric acid 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 cane sugar hydrolysate to carry out fermentation production of citric acid, this method utilizes cane sugar hydrolysate 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, a kind of cane sugar hydrolysate fermentative Production methods of citric acid of utilizing of the present invention, it adopts and earlier cultured aspergillus niger wheat bran spore is carried out seed culture in the seed culture medium that contains 9.5～10.5% concentration expressed in percentage by weight cane sugar hydrolysates, cultivates into the aspergillus niger liquid seeds; And then cultivate in the fermention medium through containing 14～16% concentration expressed in percentage by weight cane sugar hydrolysates.
Specifically, it comprises the steps:
1) flat board or slant culture: citric acid production bacterial classification aspergillus niger is inoculated into wort agar flat board or slant medium cultivation;
2) wheat bran is cultivated: the citric acid bacterial classification aspergillus niger after will cultivating then inserts a ring aspergillus niger spore, cultivates in the wheat bran seed culture medium, cultivates into the wheat bran spore;
3) seed culture: the seed culture medium that will contain 7～12% concentration expressed in percentage by weight cane sugar hydrolysates again inserts the wheat bran spore, carries out seed culture, cultivates into the aspergillus niger seed;
4) fermentation culture: cultured aspergillus niger seed is inserted in the fermention medium that contains 14～16% concentration expressed in percentage by weight cane sugar hydrolysates cultivate at last.
Wherein, citric acid production bacterial classification of the present invention is aspergillus niger Co827.
Described cane sugar hydrolysate is sucrose hydrolysate under acidic conditions or under the effect of enzyme, and it consists of: 77～81% glucose, 18～22% fructose, 0.8～1.2% sucrose.
In the step 1), the concentration of described wort is 2～4 mother-in-law U.S. degree.
Step 2) in, described wheat bran seed culture medium consists of: wheat bran 20～30%, sal epsom 0.01～0.02%, potassium primary phosphate 0.01～0.02%, 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: cane sugar hydrolysate 9.5～10.5%, corn steep liquor 0.5～1.0%, potassium primary phosphate 0.01～0.02%, ammonium sulfate 0.01～0.02%, sal epsom 0.01～0.02%, pH nature, tap water preparation.
The seed culture condition is 37 ℃ of cultivations 24～30 hours of ventilating down.
In the step 4), the consisting of of fermention medium: cane sugar hydrolysate 14～16%, corn steep liquor 0.5～1.0%, potassium primary phosphate 0.01～0.02%, ammonium sulfate 0.01～0.02%, sal epsom 0.01～0.02%, pH nature, tap water preparation.
The required carbon source of fermentation culture of the present invention is: cane sugar hydrolysate; Nitrogenous source is: ammonium sulfate, corn steep liquor.
Described leavening temperature is 35～39 ℃, and pH is 2.5～6.0, and ventilating ratio is 1: (0.1～0.6), mixing speed are 200～500rpm, cultivate 70～85 hours.
The citric acid acidity that the present invention utilizes the cane sugar hydrolysate fermentation method to obtain reaches 11～14%, and glucose acid invert ratio is greater than 89%.
The present invention utilizes cane sugar hydrolysate fermentative Production methods of citric acid, has following advantage:
1. the present invention optimizes the cane sugar hydrolysate substratum;
2. in the cane sugar hydrolysate substratum, nitrogenous source except that carbon source and nutritive element are relatively poorer, in experimentation, utilize the on-line Control means, culture media nitrogen source and nutritive element to citric acid fermentation are optimized, make cost minimization, and set up citric acid bacterial classification growth metabolism process and fermenting process modeling and control techniques;
The present invention utilize glucose in the cane sugar hydrolysate and a little sugar and fructose for carbon source adds inorganic and organic nitrogen source, various nutritive element, according to microorganism in growth, metabolic different steps to the demand difference of oxygen, various nutritive element, various carbon source, nitrogenous source, carry out fermentation production of citric acid, the fermented liquid quality obviously improves, improved the saccharic acid metabolic rate, shortened fermentation period, the citric acid fermented acidity of cane sugar hydrolysate reaches 11～14% in the fermented liquid, fermentation period 70～85 hours, glucose acid invert ratio is greater than 89%; The exploitation of opening up citric acid production raw material variation, especially foreign market had positive effect.
Embodiment
Following examples are used to illustrate the present invention, but are not used for limiting the scope of the invention.
Embodiment 1
Earlier citric acid production bacterial classification aspergillus niger spore Co827 is inoculated on wort (concentration the is 4 mother-in-law U.S. degree) agar plate and cultivates; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating to insert a ring aspergillus niger spore to containing wheat bran 30% under sterile state, 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 is linked into and contains cane sugar hydrolysate 10.5% (it is sucrose hydrolysate under acidic conditions or under the effect of enzyme, it consists of: 77% glucose, 1% sucrose, 22% fructose), corn steep liquor 0.6%, potassium primary phosphate 0.012%, 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.After treating that liquid seeds is cultivated, inoculate and contain cane sugar hydrolysate 14.3%, corn steep liquor 0.5%, potassium primary phosphate 0.011%, ammonium sulfate 0.01% is cultivated in the 5L ferment tank substratum of sal epsom 0.012%, temperature is 37 ℃, pH nature, sterile air are that 1: 0.4 rate of venting feeds in the fermention medium of fermentor tank with ventilating ratio, and mixing speed is 500rpm, ventilate and cultivated 81 hours, residual reducing sugar 1.08%, citric acid acidity reaches 11.59%, glucose acid invert ratio 89.1%.
Embodiment 2
Earlier citric acid production bacterial classification aspergillus niger spore Co827 is inoculated on wort (concentration the is 4 mother-in-law U.S. degree) agar plate and cultivates; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating to insert a ring aspergillus niger spore to containing wheat bran 20% under sterile state, sal epsom 0.02% 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 is linked into and contains cane sugar hydrolysate 10.5% (it consists of: 78% glucose, 0.9% sucrose, 21.1% fructose), corn steep liquor 0.6%, potassium primary phosphate 0.015%, ammonium sulfate 0.02% is in the liquid seed culture medium of sal epsom 0.013%, the pH nature, the tap water preparation.37 ℃, ventilate and cultivated 28 hours.After treating that liquid seeds is cultivated, inoculate and contain cane sugar hydrolysate 14.7%, corn steep liquor 0.8%, potassium primary phosphate 0.015%, ammonium sulfate 0.012% is cultivated in the 5L ferment tank substratum of sal epsom 0.018%, temperature is 39 ℃, pH is 5.5, and 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 500rpm, ventilate and cultivated 79 hours, residual reducing sugar 1.03%, citric acid acidity reaches 11.99%, glucose acid invert ratio 89.4%.
Embodiment 3
Earlier citric acid production bacterial classification aspergillus niger spore Co827 is inoculated on wort (concentration the is 4 mother-in-law U.S. degree) agar plate and cultivates; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating to insert a ring aspergillus niger spore to containing wheat bran 30% under sterile state, sal epsom 0.014% 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 is linked into and contains cane sugar hydrolysate (it consists of: 78% glucose, 1.1% sucrose, 20.9% fructose) 10.5%, corn steep liquor 0.6%, potassium primary phosphate 0.01%, ammonium sulfate 0.01% is in the liquid seed culture medium of sal epsom 0.012%, the pH nature, the tap water preparation.37 ℃, ventilate and cultivated 26 hours.After treating that liquid seeds is cultivated, inoculate and contain cane sugar hydrolysate 14.4%, corn steep liquor 1.0%, potassium primary phosphate 0.014%, ammonium sulfate 0.015% is cultivated in the 5L ferment tank substratum of sal epsom 0.012%, temperature is 37 ℃, pH is 4.5, and 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 200rpm, ventilate and cultivated 76 hours, residual reducing sugar 0.85%, citric acid acidity reaches 12.95%, glucose acid invert ratio 89.2%.
Embodiment 4
Earlier citric acid production bacterial classification aspergillus niger spore Co827 is inoculated on wort (concentration the is 4 mother-in-law U.S. degree) agar plate and cultivates; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating to insert a ring aspergillus niger spore to containing wheat bran 25% under sterile state, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.02%, was cultivated 9 days for 37 ℃.To cultivate sophisticated wheat bran spore is linked into and contains cane sugar hydrolysate 10.5% (it consists of: 78.9% glucose, 1.1% sucrose, 20% fructose), corn steep liquor 0.6%, potassium primary phosphate 0.01%, ammonium sulfate 0.018% is in the liquid seed culture medium of sal epsom 0.01%, the pH nature, the tap water preparation.37 ℃, ventilate and cultivated 29 hours.After treating that liquid seeds is cultivated, inoculate and contain cane sugar hydrolysate 14.5%, corn steep liquor 0.6%, potassium primary phosphate 0.01%, ammonium sulfate 0.02% is cultivated in the 5L ferment tank substratum of sal epsom 0.01%, temperature is 39 ℃, pH is 4.0, and 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 500rpm, ventilate and cultivated 75 hours, residual reducing sugar 1.09%, citric acid acidity reaches 13.59%, glucose acid invert ratio 89.55%.
Embodiment 5
Earlier citric acid production bacterial classification aspergillus niger spore Co827 is inoculated on wort (concentration the is 2 mother-in-law U.S. degree) agar plate and cultivates; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating to insert a ring aspergillus niger spore to containing wheat bran 30% under sterile state, sal epsom 0.01% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.016%, was cultivated 10 days for 37 ℃.To cultivate sophisticated wheat bran spore is linked into and contains cane sugar hydrolysate 10% (it consists of: 79.8% glucose, 1.2% sucrose, 19% fructose), corn steep liquor 0.8%, potassium primary phosphate 0.016%, ammonium sulfate 0.02% is in the liquid seed culture medium of sal epsom 0.013%, the pH nature, the tap water preparation.37 ℃, ventilate and cultivated 30 hours.After treating that liquid seeds is cultivated, inoculate and contain cane sugar hydrolysate 15.3%, corn steep liquor 0.9%, potassium primary phosphate 0.017%, ammonium sulfate 0.01% is cultivated in the 50L ferment tank substratum of sal epsom 0.016%, temperature is 38 ℃, pH is 6.0, and 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, ventilate and cultivated 78 hours, residual reducing sugar 0.88%, citric acid acidity reaches 13.66%, glucose acid invert ratio 89.33%.
Embodiment 6
Earlier citric acid production bacterial classification aspergillus niger spore Co827 is inoculated on wort (concentration the is 4 mother-in-law U.S. degree) agar plate and cultivates; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating to insert a ring aspergillus niger spore to containing wheat bran 30% under sterile state, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.02%, was cultivated 9 days for 37 ℃.To cultivate sophisticated wheat bran spore is linked into and contains cane sugar hydrolysate 10.5% (it consists of: 80% glucose, 0.8% sucrose, 19.2% fructose), corn steep liquor 0.9%, potassium primary phosphate 0.015%, ammonium sulfate 0.01% is in the liquid seed culture medium of sal epsom 0.016%, the pH nature, the tap water preparation.37 ℃, ventilate and cultivated 26 hours.After treating that liquid seeds is cultivated, inoculate and contain cane sugar hydrolysate 16.0%, corn steep liquor 0.5%, potassium primary phosphate 0.014%, ammonium sulfate 0.02% is cultivated in the 50L ferment tank substratum of sal epsom 0.01%, temperature is 37 ℃, pH is 2.5, and 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 400rpm, ventilate and cultivated 77 hours, residual reducing sugar 1.15%, citric acid acidity reaches 13.7%, glucose acid invert ratio 89.29%.
Embodiment 7
Earlier citric acid production bacterial classification aspergillus niger spore Co827 is inoculated on wort (concentration the is 4 mother-in-law U.S. degree) agar plate and cultivates; Adopt the dull and stereotyped citric acid production bacterial classification of cultivating to insert a ring aspergillus niger spore to containing wheat bran 30% under sterile state, sal epsom 0.02% in the wheat bran seed culture medium triangular flask of potassium primary phosphate 0.01%, was cultivated 7 days for 37 ℃.To cultivate sophisticated wheat bran spore is linked into and contains cane sugar hydrolysate 9.5% (it consists of: 80% glucose, 1.2% sucrose, 18.8% fructose), corn steep liquor 0.6%, potassium primary phosphate 0.018%, ammonium sulfate 0.015% is in the liquid seed culture medium of sal epsom 0.01%, the pH nature, the tap water preparation.37 ℃, ventilate and cultivated 30 hours.After treating that liquid seeds is cultivated, inoculate and contain cane sugar hydrolysate 15.1%, corn steep liquor 0.8%, potassium primary phosphate 0.01%, ammonium sulfate 0.02% is cultivated in the 500L ferment tank substratum of sal epsom 0.019%, temperature is 37 ℃, pH is 4.0, and 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, ventilate and cultivated 74 hours, residual reducing sugar 1.01%, citric acid acidity reaches 14.2%, glucose acid invert ratio 89.7%.
Though 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 (1)
1. one kind is utilized cane sugar hydrolysate fermentative Production methods of citric acid, it is characterized in that it comprises the steps:
1) flat board or slant culture: citric acid production bacterial classification aspergillus niger Co827 is inoculated into wort agar flat board or slant medium cultivation;
2) wheat bran is cultivated: the citric acid bacterial classification aspergillus niger Co827 after will cultivating then inserts a ring aspergillus niger spore, in the wheat bran seed culture medium, cultivate, cultivate into the wheat bran spore, described wheat bran seed culture medium consists of: wheat bran 20～30%, sal epsom 0.01～0.02%, potassium primary phosphate 0.01～0.02%, pH nature, tap water preparation, described wheat bran culture condition are 37 ℃ and cultivated 7～10 days down;
3) seed culture: the seed culture medium that will contain 9.5～10.5% concentration expressed in percentage by weight cane sugar hydrolysates again inserts the wheat bran spore, carry out seed culture, cultivate into the aspergillus niger seed, described cane sugar hydrolysate consists of: 77～81% glucose, 18～22% fructose, 0.8～1.2% sucrose, consisting of of described seed culture medium: cane sugar hydrolysate 9.5～10.5%, corn steep liquor 0.5～1.0%, potassium primary phosphate 0.01～0.02%, ammonium sulfate 0.01～0.02%, sal epsom 0.01～0.02%, the pH nature, the tap water preparation, culture condition is 37 ℃ of cultivations 24～30 hours of ventilating down;
4) fermentation culture: cultured aspergillus niger seed is linked in the fermention medium that contains 14～16% concentration expressed in percentage by weight cane sugar hydrolysates cultivates at last, consisting of of described fermention medium: cane sugar hydrolysate 14～16%, corn steep liquor 0.5～1.0%, potassium primary phosphate 0.01～0.02%, ammonium sulfate 0.01～0.02%, sal epsom 0.01～0.02%, the pH nature, the tap water preparation, described leavening temperature is 35～39 ℃, and pH is 2.5～6.0, and ventilating ratio is 1: (0.1～0.6), mixing speed is 200～500rpm, cultivates 70～85 hours.
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| CN102443611B (en) * | 2010-10-13 | 2014-03-05 | 中粮生物化学（安徽）股份有限公司 | Production method of citric acid |
| CN103710397B (en) * | 2013-12-27 | 2015-09-09 | 潍坊英轩实业有限公司 | A kind of technique utilizing beta-cyclodextrin mother liquor for fermentation to prepare citric acid |
| CN104087624B (en) * | 2014-07-11 | 2017-08-08 | 江南大学 | Aspergillus niger continuously ferment production citric acid method |
| CN106381148A (en) * | 2016-08-24 | 2017-02-08 | 云南民族大学 | Preparing and using methods of an environment-friendly type soil heavy metal eluant |
Citations (1)
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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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| S. S. Tsay et al..Citric Acid Production Using Immobilized Conidia of Aspergillus niger TMB 2022.《Biotechnology and Bioengineering》.1987,第297-304页. * |
| 乌敏辰.淀粉原料发酵生产柠檬酸的研究.《山西食品工业》.1995,第13-15页. * |
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