CN102321685A - A kind of preparation method of citric acid fermentation broth - Google Patents
A kind of preparation method of citric acid fermentation broth Download PDFInfo
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- CN102321685A CN102321685A CN201110226953A CN201110226953A CN102321685A CN 102321685 A CN102321685 A CN 102321685A CN 201110226953 A CN201110226953 A CN 201110226953A CN 201110226953 A CN201110226953 A CN 201110226953A CN 102321685 A CN102321685 A CN 102321685A
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Abstract
The invention provides a kind of preparation method of citric acid fermentation broth, this method comprises: the step of sizing mixing: adjusting slurry is mixed with starchy material, obtain farinaceous size; Said adjusting slurry is for containing the lemon aqueous acid; Said contain the lemon aqueous acid contain come from fermentation step obtain producing in the process that citric acid fermentation broth purifies in and waste liquid, the weight ratio of said starchy material and said adjusting slurry is 1: 1.86-3; Enzymolysis step: under enzymatic hydrolysis condition, the farinaceous size that the step of sizing mixing is obtained mixes with enzyme and carries out enzymolysis, obtains enzymolysis product; Fermentation step: under the condition that generates Hydrocerol A, the enzymolysis product that enzymolysis step is obtained ferments, and obtains citric acid fermentation broth; Fermentation termination is the residual reducing sugar content 1-10g/L of fermented liquid.Method of the present invention can reduce the process water amount greatly, shortens the production cycle, improves the production efficiency of Hydrocerol A, has also reached to save the purpose of the energy, and has obtained the high concentration of citric fermented liquid.
Description
Invention field
The present invention relates to a kind of preparation method of citric acid fermentation broth.
Background of invention
Hydrocerol A is important industrial organic acid, is called as first edible acidic flavoring agent, in foodstuffs industry, is widely used, and also is widely used at medicine, beauty treatment, makeup, washing composition, industrial circle simultaneously.Since 1784, Sweden chemist Scheel extracted Hydrocerol A and crystallization first and goes out after the solid citric acid from lemon juice, constantly studied exploration through external scientist, had realized the tank fermentation method industriallization, thereby had realized the scale operation Hydrocerol A.
China's citric acid industry is started in the fifties in last century, makes joint efforts through the several generations, and the Hydrocerol A state of the art has been in rank first, has realized developing rapidly of citric acid industry.The preparation of Hydrocerol A generally comprises: amylaceous raw material is carried out pre-treatment and fermentation obtains fermentation clear liquid, and further purifying citric acid from fermentation clear liquid, and carry out aftertreatment.
At present; Industry top fermentation method is produced Hydrocerol A a large amount of water commonly used and is come the W-Gum raw material is sized mixing; And because citric acid fermented used starchy material proportion in the citric acid fermentation cost is bigger, so that the citric acid fermentation endpoint selects to control the content of residual reducing sugar usually is low as far as possible.Do like this and can reduce citric acid production grain consumption on the one hand, can reduce the COD value of citric acid wastewater on the one hand.But this method can cause the waste of water resources; The more important thing is,, can cause the fermentative prodn cycle stretch-out for the content of controlling residual reducing sugar is low as far as possible; The problem that energy consumption is high; And also can produce the waste water of a large amount of contaminate environment, in addition, also exist certain difficulty on the Hydrocerol A acidity in improving fermented liquid.
Summary of the invention
In order to solve the problems referred to above that exist in the prior art, goal of the invention of the present invention is to provide a kind of water saving, high-level efficiency, less energy-consumption, the low preparation method who pollutes and can access the citric acid fermentation broth of peracid degree Hydrocerol A.
For achieving the above object, the invention provides a kind of preparation method of citric acid fermentation broth, this method comprises:
The step of sizing mixing: adjusting slurry is mixed with starchy material, obtain farinaceous size; Said adjusting slurry is for containing the lemon aqueous acid; Said contain the lemon aqueous acid contain come from fermentation step obtain producing in the process that citric acid fermentation broth purifies in and waste liquid, the weight ratio of said starchy material and said adjusting slurry is 1: 1.86-3;
Enzymolysis step: under enzymatic hydrolysis condition, the farinaceous size that the step of sizing mixing is obtained mixes with enzyme and carries out enzymolysis, obtains enzymolysis product;
Fermentation step: under the condition that generates Hydrocerol A, the enzymolysis product that enzymolysis step is obtained ferments, and obtains citric acid fermentation broth; Fermentation termination is that the residual reducing sugar content of fermented liquid is 1-10g/L.
The present invention has following advantage: the water consumption is low: owing to being back to the allotment farinaceous size with waste liquid in producing in the citric acid fermentation broth purification process, therefore, can reduce the consumption of the water of sizing mixing; Throughput rate is high: through content that improves starchy material in the farinaceous size and the judgement criteria of passing through to change fermentation termination, make fermentation period will shorten 10%-20% than with the alap standard of residual sugar content, Gu and fermentation production efficiency is greatly improved; Fermented liquid terminal point acidity is high: because high sugar-fermenting can influence residual sugar, return the problem of joining better solution residual sugar with waste liquid in this law employing, thus can improve fermentation sugar just, and then raising fermented liquid terminal point acidity.Simultaneously since in the fermentation with the recycling of waste liquid, minimum to the influence of sugar consumption, and greatly degree reduces and drains in the environmental protection and waste liquid COD, alleviates environmental protection pressure.Help calcium salt method, ISEP methods such as (continuous ionic exchange chromatographies) extraction Hydrocerol A.Energy consumption is low: since the fermentation production efficiency raising, the air that unit product consumed, and power cost also has the reduction of certain amplitude.
Embodiment
Preparing method according to citric acid fermentation broth provided by the invention; Wherein, In the said step of sizing mixing; Said adjusting slurry is for containing the lemon aqueous acid, and the said lemon aqueous acid that contains contains and comes from obtaining in the said fermentation step producing in the process that citric acid fermentation broth purifies and waste liquid.Employing contains the lemon aqueous acid can be practiced thrift the water of sizing mixing originally as adjusting slurry consumption; To realize the purpose of waste water recycling, conserve water resource; The more important thing is, with also containing partial reduction sugar in obtaining in the said fermentation step producing in the process that citric acid fermentation broth purifies with in the waste liquid.Therefore, on the one hand, can again reducing sugar and the starchy material that this part is not fermented be mixed for further enzymolysis and fermentation through the reuse of waste water; Thereby the residual reducing sugar that need not wait until fermentation termination is zero or just stops fermentation near zero the time; Thereby improved fermentation efficiency greatly, shortened the production cycle, can also save energy consumption; Can also increase the amount of the reducing sugar that can be used for fermenting on the other hand, to reach the purpose that improves Hydrocerol A acidity.Moreover the said lemon aqueous acid that contains contains in obtaining in the said fermentation step producing in the process that citric acid fermentation broth purifies and waste liquid, can reduce discharged waste water, alleviates the pollution to environment.
According to the present invention; In the said step of sizing mixing, contain the lemon aqueous acid and be and contain from obtaining in the said fermentation step producing in the process that citric acid fermentation broth purifies and waste liquid, for example; Can directly adopt said in and waste liquid, or in said with the mixture of waste liquid and water.For in have no particular limits with the blending ratio of waste liquid and water, but consider from the purpose of water saving, of the present inventionly contain in the preferred all uses of lemon aqueous acid and waste liquid.
Preparing method according to citric acid fermentation broth provided by the invention; Wherein, in the said step of sizing mixing, in Citric acid monohydrate Food grade; The said content that contains Hydrocerol A in the lemon aqueous acid can change in the larger context; Because in order to reach the purpose of waste water recycling, said contain the lemon aqueous acid contain with fermentation step obtain producing in the process that citric acid fermentation broth purifies in and waste liquid, therefore; In Citric acid monohydrate Food grade, 1 liter of said content that contains Hydrocerol A in the lemon aqueous acid can restrain for 1-3; Under the preferable case, 1 liter of said content that contains Hydrocerol A in the lemon aqueous acid is the 1.5-3 gram; More preferably under the situation, 1 liter of said content that contains Hydrocerol A in the lemon aqueous acid is the 2-2.5 gram.
According to the preparation method of citric acid fermentation broth provided by the invention, wherein, in the said step of sizing mixing, preferred, the weight ratio of said starchy material and said adjusting slurry is 1: 1.86-3; More preferably, the weight ratio of said starchy material and said adjusting slurry is 1: 2.13-2.7; Most preferably, the weight ratio of said starchy material and said adjusting slurry is 1: 2.3-2.57.
Here; The standard of judging owing to the selection fermentation termination in the prior art is that residual reducing sugar is zero or acidity no longer increases, the time that the indefinite prolongation of use high density slurries possibility is reached home, therefore; The consumption of starchy material is lower in the farinaceous size; Weight ratio like starchy material and water is 1: about 3.2-4, be that the residual reducing sugar content of fermented liquid is 1-10g/L because the present invention only need control fermentation termination, therefore; Correspondingly can use the farinaceous size of higher concentration to ferment, thereby can access the fermented liquid of peracid degree.
According to the preparation method of citric acid fermentation broth provided by the invention, wherein, in the step of sizing mixing, the pH value of said farinaceous size is preferably 5.0-6.5; 5.5-6.0 more preferably.
According to the preparation method of citric acid fermentation broth provided by the invention, wherein, preferred, the residual reducing sugar of the fermentation termination of fermented liquid is 1-10g/L; More preferably, the residual reducing sugar of fermentation termination is less than 2-9g/L.Here, raw material accounts for larger proportion in cost in the prior art, is that residual reducing sugar is zero or acidity no longer increases in order to make raw material fully obtain fermentation, to save the grain consumption in the fermentation termination Standard Selection therefore, has so just prolonged fermentation period, inefficiency; The present invention will be owing to will use in the fermented liquid and waste liquid returns and joins; In with waste liquid in unconverted reducing sugar get back in the raw slurry once more; Get into the fermentation circulation, the residual reducing sugar that therefore need not obtain in the fermented liquid transforms fully, thereby can shorten the production cycle; Improve efficient, can also save energy consumption.
Among the present invention, with obtain in the said fermentation step citric acid fermentation broth purify obtain in the method for waste liquid can be for well known to a person skilled in the art method, for example; With obtain in the calcium salt method purifying citric acid fermentation clear liquid process once in waste liquid and/or secondary in and waste liquid, can obtain through following method: citric acid fermentation broth is carried out press filtration to remove thalline, obtain fermentation clear liquid after the press filtration; Under 75 ℃, lime carbonate is slowly joined in the citric acid fermentation clear liquid, when control pH value is 3.8-4.1; Stop to add lime carbonate, reacted 4.8-5.1 minute, after the layering of question response liquid; Filter, the filtrating that obtains is in once and waste liquid.
According to the present invention, the preparation method of said farinaceous size can adopt the method that well known to a person skilled in the art various routines, for example, starchy material is pulverized, and the product after pulverizing is mixed obtaining farinaceous size with water.Condition and mode that starchy material is pulverized have no particular limits, as long as can make starchy material fully broken, under the preferable case, the particle diameter of 50-60 weight % is less than 0.1cm in the particle of the product after the pulverizing that obtains.Said starchy material can be the known various raw materials that contain starch that can be used for enzymolysis, fermentative prepn Hydrocerol A of ability, for example, can be selected from corn, potato class (like cassava), wheat and the Chinese sorghum one or more.
According to the present invention, be conventionally known to one of skill in the art with the method for farinaceous size enzymolysis.For example, farinaceous size is mixed with microbes producing cellulase and/or enzyme, insulation is accomplished enzymolysis and is obtained enzymolysis product under the growth temperature of microbes producing cellulase and/or the great-hearted temperature of enzyme.
According to the present invention, being controlled to be of the terminal point pol of said enzymolysis product is conventionally known to one of skill in the art, and the terminal point pol of preferred enzyme hydrolysis products is 15-25g/L, more preferably 19-22g/L.
The enzymatic hydrolysis condition of the preparation Hydrocerol A that 00,000,000,000,000,000,000,000,001,111,111,111,111,111,111,111,111,111,111 1111111111111111111111 field technique personnel are known; For example; The temperature of said enzymolysis can be diastatic any optimum temperature; Be generally 70-98 ℃, more preferably 80-90 ℃; The longer the better on the time theory of enzymolysis, considers plant factor, and the time of preferred said enzymolysis is 60-180 minute, more preferably 100-130 minute; The pH value of said enzymolysis can be generally 5.0-6.5 for diastatic the righttest any action pH, and more preferably the pH value is 5.3-6.3, and most preferably the pH value is 5.5-6.0.
Because microorganism growth can produce by product, the therefore preferred enzyme that directly adds.The consumption of said enzyme is The more the better; From cost consideration, preferably with the dry weight basis of every gram starchy material, said diastatic consumption is the 10-60 enzyme activity unit; More preferably with the dry weight basis of every gram starchy material, said diastatic consumption is the 15-50 enzyme activity unit.
Being defined as of the enzyme activity unit of enzyme according to the invention: be 6.0 in the pH value, temperature is that 70 ℃ the required enzyme amount of bar 111 grape sugar is an enzyme activity unit.
Said enzyme comprises glycase.
Glycase is meant the general name of class of enzymes that can the starch-splitting glycosidic link, and said glycase generally comprises AMS, beta-amylase, saccharifying enzyme and isoamylase.Enzyme according to the invention comprises glycase.
AMS is claimed starch 1 again, the 4-dextrinase, and it can cut the inner α-1 of starch chain at random, brokenly, and the 4-glycosidic link is hydrolyzed to starch SANMALT-S, contains the oligosaccharides of 6 glucose units and has the oligosaccharides of side chain.The mikrobe that produces this enzyme mainly has Bacillus subtilus, black mold, aspergillus oryzae and head mold.
Beta-amylase is claimed starch 1 again, and 4-maltoside enzyme can cut 1 from the starch molecule non reducing end, and the 4-glycosidic link generates SANMALT-S.The product that this enzyme acts on starch is SANMALT-S and limit dextrin.This enzyme is mainly produced by aspergillus, head mold and endomyces.
Saccharifying enzyme is claimed starch α-1 again, the 4-glucuroide, and this enzyme acts on the non reducing end of starch molecule, is unit with glucose, acts on the α-1 in the starch molecule successively, and the 4-glycosidic link generates glucose.The product that saccharifying enzyme 0 acts on behind the pulullan has glucose and has α-1, the oligosaccharides of 6-glycosidic link; The product that acts on after the amylose starch almost all is a glucose.This enzyme produces bacterium mainly to be black mold (left U.S. aspergillus, Aspergillus awamori), head mold (snow-white enzyme, De Shi head mold), to intend endomyces, monascus.
Isoamylase is claimed starch α-1 again, and 6-glucuroide, branching enzyme, this enzyme act on the α-1 at amylopectin molecule branching-point place, and the 6-glycosidic link is with whole side chain cutting-out the becoming amylose starch of amylopectin.It mainly is to dislike bacteriums such as gas bacillus, genus bacillus and some false monospore bacillus that this enzyme produces bacterium.
According to the present invention; Under the condition that generates Hydrocerol A; Enzymolysis product is fermented, and the method that obtains citric acid fermentation broth can be for well known to a person skilled in the art method, for example; Generating under the condition of Hydrocerol A, black mold is being seeded in the fermention medium that contains enzymolysis product ferments.
According to the present invention, said fermention medium can be the conventional fermention medium that is used to produce Hydrocerol A in this area.For example, can enzymolysis product be carried out solid-liquid separation and obtain enzymolysis residue and enzymolysis clear liquid, and with the enzymolysis clear liquid be mixed into certain proportion preparation fermention medium without the enzymolysis product of solid-liquid separation.For the carrying out that is easier to ferment, under the preferable case, be benchmark with the culture volume, the content of said enzymolysis clear liquid is 70-90 volume %, the content of said enzymolysis product without solid-liquid separation is 10-30 volume %.More preferably under the situation, said enzymolysis clear liquid content is 75-85 volume %, is 15-25 volume % without the filter enzymolysis product volume of solid-liquid separation.
According to method of the present invention; This method can also comprise adds nitrogenous source in fermention medium; The time of adding nitrogenous source accomplished in the preceding 5 hours time period to fermentation after black mold is seeded to fermention medium in 24 hours, and the effect of adding nitrogenous source is growth and metabolism in order better to keep black mold.Wherein, The kind of said nitrogenous source is conventionally known to one of skill in the art, and for example, said nitrogenous source can be in urea, ammonium sulfate and an ammonium nitrate one or more; The add-on of said nitrogenous source can in very large range change; Under the preferable case, be benchmark with the gross weight of said fermention medium, the add-on of said nitrogenous source is 0.01-0.1 weight %.
According to the present invention, the inoculum size of said black mold can in very large range change, and under the preferable case, is benchmark with every liter of fermention medium, and the inoculum size of black mold is 5 * 10
4-5 * 10
5Individual spore, more preferably 2.5 * 10
5-3.5 * 10
5Individual spore.
The number of said spore can be measured by means commonly known in the art, for example, counts through blood counting chamber.Specifically can measure through following method: get the wheat bran of a flask culture black mold, pour in the bottle 0.5% tween-80 into constant volume to V1=500mL, putting into rotor simultaneously, to be stirred to suspension even.Get the even spore liquid dilution of V2=5mL and be settled to V3=100mL; Ultra-sonic dispersion 10 minutes is poured into and is made it even with magnetic stirring apparatus in the beaker, and the blood counting chamber counting is adopted in sampling; Counting draws spore suspension spore count N after every milliliter of dilution, then every bottle of wheat bran spore count=V1 * (N * V3/V2).
According to the present invention, the condition of fermenting in the said fermentation step can in very large range change, and obtains Hydrocerol A as long as can ferment; For example; The condition of fermentation comprises: the temperature of fermentation is 25-45 ℃, and the pH value is 1-7, and tank pressure is 0.05-0.1Mpa; Air flow is 0.15-0.5V/Vmin, and the time of fermentation is 45-75 hour; Under the preferred situation, the condition of said fermentation comprises: the temperature of fermentation is 30-40 ℃, and the pH value is 2-4, and tank pressure is 0.06-0.09Mpa, and air flow is 0.2-0.3V/Vmin, and the time of said fermentation is 55-60 hour.
Term " air flow " is generally with ventilation expression recently, and usually recently to represent (V/Vmin) through the volume of air of unit volume nutrient solution in the PM, for example ventilation is than being 1: 0.05-0.5, the abbreviation air flow is the 0.05-0.5 volume: volume minute.
The present invention's employed black mold that ferments can be black mold solid preparation or the aspergillus niger strain that is purchased; For example, black mold Co827 (Xinli Industrial Microbe Science and Technology Co Ltd, Shanghai), black mold T01 (Tianjin industrial microorganism institute) and Aspergillus niger strain (Institute of Micro-biology of the Chinese Academy of Sciences).
Said black mold can adopt conventional method inoculation, for example, in being seeded to fermention medium before, said black mold is handled through seed culture, afterwards the seed liquor that obtains is joined in the fermention medium.The degree of black mold seed culture can be measured through sampling sediments microscope inspection, acid test and pH and observe the growth of black mold, when pH 2.0-2.5, acidity 0.5-2.0 weight %, bacterium ball size evenly, mycelia is sturdy stops to cultivate when stretching out.
According to the present invention, the preparation method of said black mold nutrient solution has no particular limits, as long as the nutrient solution that obtains can be applicable to the cultivation of black mold.
Among the present invention, the inoculum size of said black mold can in very large range change, and under the preferable case, black mold is seeded in the black mold nutrient solution cultivates, and the concentration of black mold is in every milliliter of black mold nutrient solution 1 * 10 in the black mold nutrient solution of inoculation back
5-3 * 10
5Individual spore.
For can be when black mold be carried out seed culture; For black mold competent nutrient is provided better; Can also in the black mold nutrient solution, add nitrogenous source, be benchmark with the gross weight of said black mold nutrient solution, and the add-on of said nitrogenous source can be 0.01-0.1 weight %.
According to the present invention; The culture condition of said black mold can in very large range change; For example said culture condition can comprise: the temperature of cultivation can be 30-45 ℃; The pH value can be 1-7, and air flow can be the 0.03-0.2 volume: (volume minute), and the time of cultivation can be 18-30 hour; Under the preferred situation, said culture condition can comprise: the temperature of cultivation can be 35-40 ℃, and the pH value can be 2-4, and air flow can be the 0.06-0.1 volume: (volume minute), the time of said cultivation can be 20-27 hour.
The equipment of said cultivation is conventionally known to one of skill in the art, for example, can use fermentor tank to cultivate.
According to the present invention, the measuring method that the terminal point pol is can be a whole bag of tricks well known in the art, and preferred, the Fehling method is with reference to GB GBT50099-2008.
According to the present invention, residual reducing sugar and the measuring method that per hour increases acid can be the whole bag of tricks well known in the art, and be preferred, the Fehling method with reference to GB GBT50099-2008 and Na (OH) volumetry with reference to GB GBT 8269-2006.
According to the present invention, the measuring method of fermented liquid terminal point acidity can be a whole bag of tricks well known in the art, and preferred, Na (OH) volumetry is with reference to GB GBT 8269-2006.
Further illustrate in greater detail the present invention through the following example, embodiment is unrestricted the present invention for explanation.Any those of ordinary skill can be understood these embodiment and not limit the present invention in any way in this area, modification that can be suitable and without prejudice to essence of the present invention with depart from scope of the present invention.
Embodiment 1
Present embodiment is used to explain the preparation of citric acid fermentation broth provided by the invention.
1) pulverizing of raw material: the corn of results is stewing in the hot water tank profit; Water cut until corn is 15 weight %; Then through kibbler (Jiangsu MuYang Group, Ltd.; The 968-3 type) pulverizes, obtain starchy material crushed products (particle diameter of 50 weight % is less than 0.1cm in the pulverized particles);
2) size mixing: use from the step 8) and waste liquid (in 1 liter with waste water in the aqueous solution in the content of Hydrocerol A be 2.2 grams) mix with starchy material to size mixing as adjusting slurry and obtain farinaceous size; The weight ratio of said starchy material and said adjusting slurry is 1: 2.57, and the pH value of farinaceous size is 5.5;
3) farinaceous size that enzymolysis: 2) obtains and glycase (Novozymes Company; AMS, equal glycase for this reason in the embodiment of the invention) mixing, is to carry out enzymolysis 100 minutes under 5.5 the condition at 90 ℃, pH; With respect to the product after every gram pulverizing; Diastatic consumption is 40 enzyme activity units, obtains enzymolysis product, and the terminal point pol is 20.2g/L;
4) enzymolysis product of the 80 weight % that filtration: with 3) obtain is isolated enzymolysis clear liquid and enzymolysis residue through carrying out press filtration with the fluid pressure type plate-and-frame filter press;
5) preparation fermention medium: join in the fermentor tank of 300L with the enzymolysis clear liquid with after the enzymolysis product of solid-liquid separation mixes with 85: 15 volume ratio; Obtain fermention medium after the sterilization; Carbon source content is 19.6 weight % in the said fermention medium; Nitrogenous source content is 0.10 weight %, and phosphorus source content is 350ppm;
6) with the part enzymolysis product thin up in the step 3) to 1/10 of weight, obtain nutrient solution, nutrient solution is dropped into seeding tank; Be heated to 121 ℃ of sterilizations, keep after 30 minutes fast cooling to 36 ℃, insert aspergillus niger strain (black mold T01; Tianjin industrial microorganism institute; Equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of nutrient solution is a benchmark, inoculation 3 * 10
5Individual spore), be 3 in the pH value, temperature is to carry out spawn culture under 36 ℃, the aeration condition of 0.4V/Vmin; Measure through sampling sediments microscope inspection, acid test and pH the growth of black mold observed, after 27 hours, when pH 2.0, acidity 10g/L, bacterium ball size evenly, mycelia is sturdy when stretching out, stop to cultivate;
7) fermentation: 5) insert 6 in the substratum of gained) in cultivate aspergillus niger strain (black mold Co827, Tianjin industrial microorganism institute, equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of fermention medium is a benchmark, inoculation 3 * 10
5Individual spore), and the ventilation oxygen supply, be 4 in original ph, tank pressure is about 0.08Mpa, air flow 0.15V/Vmin, under the condition of 37 ℃ of culture temperature, cultivate 55h after, as residual reducing sugar 3g/L, stop jar, terminal point acidity is 177g/L;
8) in the preparation of waste liquid: with 7) citric acid fermentation broth that obtains carries out press filtration to remove thalline, obtains fermentation clear liquid after the press filtration, under 75 ℃; Lime carbonate is slowly joined in the citric acid fermentation clear liquid (content of Hydrocerol A is 179 grams in every liter of citric acid fermentation clear liquid), and control pH value is 4.8 o'clock, stops to add lime carbonate; Reacted 60 minutes; After the layering of question response liquid, filter, obtain filtrating in once and waste liquid.
Embodiment 2
Present embodiment is used to explain the preparation of citric acid fermentation broth provided by the invention.
1) peeling of raw material and pulverizing: the corn of results is stewing in the hot water tank profit; Water cut until corn is 15 weight %; Then through kibbler (Jiangsu MuYang Group, Ltd.; The 968-3 type) pulverizes, obtain starchy material (particle diameter of 52 weight % is less than 0.1cm in the pulverized particles);
2) size mixing: use from the step 8) and waste liquid (in 1 liter with waste water in the aqueous solution in the content of Hydrocerol A be 2.3 grams) with former water allotment adjusting slurry; The consumption of former water accounts for 50 weight % of adjusting slurry; Said adjusting slurry mixed with starchy material to size mixing obtain farinaceous size; The weight ratio of said starchy material and said adjusting slurry is 1: 2.3, and the pH value of farinaceous size is 6.0;
3) add glycase (Novozymes Company in the farinaceous size that enzymolysis: 2) obtains; AMS, equal glycase for this reason in the embodiment of the invention) carry out enzymolysis, the product after pulverizing with respect to every gram; Diastatic consumption is 40 enzyme activity units; Be enzymolysis 120 minutes under 5.0 the condition at 95 ℃, pH with this mixture, obtain enzymolysis product, the terminal point pol is 21.0g/L;
4) filtering: 3) 80% the enzymolysis product that obtains is through carrying out press filtration with the fluid pressure type plate-and-frame filter press, isolates enzymolysis clear liquid and enzymolysis residue;
5) preparation fermention medium, with the enzymolysis stillness of night with do not filter enzymolysis product and join in the fermentor tank of 300L after with the mixing of 75: 25 volume ratio, obtain fermention medium; Wherein, Carbon source content is 20.4 weight %, and nitrogenous source content is 0.12 weight %, and phosphorus source content is 400ppm;
6) with the part enzymolysis product thin up in the step 3) to 1/10 of weight, obtain nutrient solution, nutrient solution is dropped into seeding tank; Be heated to 121 ℃ of sterilizations, keep after 30 minutes fast cooling to 36 ℃, insert aspergillus niger strain (black mold T01; Tianjin industrial microorganism institute; Equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of nutrient solution is a benchmark, inoculation 3 * 10
5Individual spore), be 3 in the pH value, temperature is to carry out spawn culture under 36 ℃, the aeration condition of 0.4V/Vmin; Measure through sampling sediments microscope inspection, acid test and pH the growth of black mold observed, after 22 hours, when pH 2.0, acidity 10g/L, bacterium ball size evenly, mycelia is sturdy when stretching out, stop to cultivate;
7) fermentation: 5) insert 6 in the substratum of gained) in cultivate aspergillus niger strain (black mold Co827, Tianjin industrial microorganism institute, equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of fermention medium is a benchmark, inoculation 3 * 10
5Individual spore), and the ventilation oxygen supply, be 3.5 in original ph, tank pressure is about 0.08Mpa, air flow 0.15V/Vmin, under the condition of 37 ℃ of culture temperature, cultivate 60h after, as residual reducing sugar 3g/L, stop jar, terminal point acidity is 183g/L.
8) in the preparation of waste liquid: with 7) citric acid fermentation broth that obtains carries out press filtration to remove thalline, obtains fermentation clear liquid after the press filtration, under 75 ℃; Lime carbonate is slowly joined in the citric acid fermentation clear liquid (content of Hydrocerol A is 181 grams in every liter of citric acid fermentation clear liquid), and control pH value is 4.8 o'clock, stops to add lime carbonate; Reacted 60 minutes; After the layering of question response liquid, filter, the filtrating that obtains is in once and waste liquid.
Embodiment 3
Present embodiment is used to explain the preparation of citric acid fermentation broth provided by the invention.
1) peeling of raw material and pulverizing: the corn of results is stewing in the hot water tank profit; Water cut until corn is 15 weight %; Then through kibbler (Jiangsu MuYang Group, Ltd.; The 968-3 type) pulverizes, obtain starchy material (particle diameter of 55 weight % is less than 0.1cm in the pulverized particles);
2) size mixing: with in the fermented liquid and waste liquid (in 1 liter with waste water in the aqueous solution in the content of Hydrocerol A be 2.5 grams) with former water allotment adjusting slurry; The consumption of former water accounts for 40 weight % of adjusting slurry; With the said adjusting slurry moon starchy material mix to size mixing and obtain farinaceous size; The weight ratio of said starchy material and said adjusting slurry is 1: 2.45, and the pH value of farinaceous size is 5.8;
3) add glycase (Novozymes Company, AMS, equal glycase for this reason in the embodiment of the invention) in the slurries that enzymolysis: 2) obtain and carry out enzymolysis; With respect to the product after every gram pulverizing; Diastatic consumption is 50 enzyme activity units, gets into injector, is enzymolysis 100 minutes under 5.5 the condition at 95 ℃, pH; Obtain enzymolysis product, the terminal point pol is 21.0g/L;
4) filtering: 3) 80% the enzymolysis product that obtains is through carrying out press filtration with the fluid pressure type plate-and-frame filter press, isolates enzymolysis clear liquid and enzymolysis residue;
5) preparation fermention medium, with the enzymolysis stillness of night with do not filter enzymolysis product and join in the fermentor tank of 300L after with the mixing of 8: 2 volume ratio, obtain fermention medium, wherein, carbon source content is 20.3, nitrogenous source content is 0.11%, phosphorus source content is 350ppm;
6) with the part enzymolysis product thin up in the step 3) to 1/10 of weight, obtain nutrient solution, nutrient solution is dropped into seeding tank; Be heated to 121 ℃ of sterilizations, keep after 30 minutes fast cooling to 36 ℃, insert aspergillus niger strain (black mold T01; Tianjin industrial microorganism institute; Equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of nutrient solution is a benchmark, inoculation 3 * 10
5Individual spore), be 3.5 in original ph, temperature is to carry out spawn culture under 36 ℃, the aeration condition of 0.4V/Vmin; Measure through sampling sediments microscope inspection, acid test and pH the growth of black mold observed, after 23 hours, when pH 2.0, acidity 10g/L, bacterium ball size evenly, mycelia is sturdy when stretching out, stop to cultivate;
7) fermentation: 5) insert 6 in the substratum of gained) in cultivate aspergillus niger strain (black mold Co827, Tianjin industrial microorganism institute, equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of fermention medium is a benchmark, inoculation 3 * 10
5Individual spore), and the ventilation oxygen supply, the pH value is 2, tank pressure is about 0.08Mpa, air flow 0.15V/Vmin, under the condition of 37 ℃ of culture temperature, cultivate 57h after, as residual reducing sugar 5g/L, stop jar, terminal point acidity is 185g/L.
8) in the preparation of waste liquid: with 7) citric acid fermentation broth that obtains carries out press filtration to remove thalline, obtains fermentation clear liquid after the press filtration, under 75 ℃; Lime carbonate is slowly joined in the citric acid fermentation clear liquid (content of Hydrocerol A is 183 grams in every liter of citric acid fermentation clear liquid), and control pH value is 4.8 o'clock, stops to add lime carbonate; Reacted 60 minutes; After the layering of question response liquid, filter, the filtrating that obtains is in once and waste liquid.
Embodiment 4
Present embodiment is used to explain the preparation of citric acid fermentation broth provided by the invention.
1) peeling of raw material and pulverizing: the corn of results is stewing in the hot water tank profit; Water cut until corn is 15 weight %; Then through kibbler (Jiangsu MuYang Group, Ltd.; The 968-3 type) pulverizes, obtain starchy material (particle diameter of 57 weight % is less than 0.1cm in the pulverized particles);
2) size mixing: with in the fermented liquid and waste liquid (in 1 liter with waste water in the aqueous solution in the content of Hydrocerol A be 2.3 grams) with former water allotment adjusting slurry; The consumption of former water accounts for 60 weight % of adjusting slurry; Said adjusting slurry mixed with starchy material to size mixing obtain farinaceous size; The weight ratio of said starchy material and said adjusting slurry is 1: 2.13, and the pH value of farinaceous size is 6.3;
3) add glycase (Novozymes Company, AMS, equal glycase for this reason in the embodiment of the invention) in the slurries that enzymolysis: 2) obtain and carry out enzymolysis; With respect to the product after every gram pulverizing; Diastatic consumption is 40 enzyme activity units, gets into injector, is enzymolysis 130 minutes under 6 the condition at 90 ℃, pH; Obtain enzymolysis product, the terminal point pol is 22.0g/L;
4) filtering: 3) 80% the enzymolysis product that obtains is through carrying out press filtration with the fluid pressure type plate-and-frame filter press, isolates enzymolysis clear liquid and enzymolysis residue;
5) preparation fermention medium, with the enzymolysis clear liquid with do not filter enzymolysis product and join in the fermentor tank of 300L after with the mixing of 7: 3 volume ratio, obtain fermention medium; Wherein, Carbon source content is 21.0 weight %, and nitrogenous source content is 0.13 weight %, and phosphorus source content is 450ppm;
6) with the part enzymolysis product thin up in the step 3) to 1/10 of weight, obtain nutrient solution, nutrient solution is dropped into seeding tank; Be heated to 121 ℃ of sterilizations, keep after 30 minutes fast cooling to 36 ℃, insert aspergillus niger strain (black mold T01; Tianjin industrial microorganism institute; Equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of nutrient solution is a benchmark, inoculation 3 * 10
5Individual spore), be 3 in the pH value, temperature is to carry out spawn culture under 36 ℃, the aeration condition of 0.4V/Vmin; Measure through sampling sediments microscope inspection, acid test and pH the growth of black mold observed, after 20 hours, when pH 2.0, acidity 10g/L, bacterium ball size evenly, mycelia is sturdy when stretching out, stop to cultivate;
7) fermentation: 5) insert 6 in the substratum of gained) in cultivate aspergillus niger strain (black mold Co827, Tianjin industrial microorganism institute, equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of fermention medium is a benchmark, inoculation 3 * 10
5Individual spore), and the ventilation oxygen supply, the pH value for initial 4.0, tank pressure are about 0.09Mpa, air flow 0.5V/Vmin, under the condition of 37 ℃ of culture temperature, cultivate 61h after, as residual reducing sugar 7g/L, terminal point acidity is 186g/L.
8) in the preparation of waste liquid: with 7) citric acid fermentation broth that obtains carries out press filtration to remove thalline, obtains fermentation clear liquid after the press filtration, under 75 ℃; Lime carbonate is slowly joined in the citric acid fermentation clear liquid (content of Hydrocerol A is 182 grams in every liter of citric acid fermentation clear liquid), and control pH value is 4.8 o'clock, stops to add lime carbonate; Reacted 60 minutes; After the layering of question response liquid, filter, the filtrating that obtains is in once and waste liquid.
Embodiment 5
Present embodiment is used to explain the preparation of citric acid fermentation broth provided by the invention.
1) peeling of raw material and pulverizing: the corn of results is stewing in the hot water tank profit; Water cut until corn is 15 weight %; Then through kibbler (Jiangsu MuYang Group, Ltd.; The 968-3 type) pulverizes, obtain starchy material (particle diameter of 60 weight % is less than 0.1cm in the pulverized particles);
2) size mixing: with in the fermented liquid and waste liquid (in 1 liter with waste water in the aqueous solution in the content of Hydrocerol A be 2.0 grams) with former water allotment adjusting slurry; The consumption of former water accounts for 40 weight % of adjusting slurry; Said adjusting slurry mixed with starchy material to size mixing obtain farinaceous size; The weight ratio of said starchy material and said adjusting slurry is 1: 2.7, and the pH value of farinaceous size is 5.3;
3) add glycase (Novozymes Company, AMS, equal glycase for this reason in the embodiment of the invention) in the slurries that enzymolysis: 2) obtain and carry out enzymolysis; With respect to the product after every gram pulverizing; Diastatic consumption is 40 enzyme activity units, gets into injector, is enzymolysis 100 minutes under 5.0 the condition at 80 ℃, pH; Obtain enzymolysis product, the terminal point pol is 19.0g/L;
4) filtering: 3) 80% the enzymolysis product that obtains is through carrying out press filtration with the fluid pressure type plate-and-frame filter press, isolates enzymolysis clear liquid and enzymolysis residue;
5) preparation fermention medium, with the enzymolysis clear liquid with do not filter enzymolysis product and join in the fermentor tank of 300L after with the mixing of 9: 1 volume ratio, obtain fermention medium; Wherein, Carbon source content is 18.4 weight %, and nitrogenous source content is 0.07 weight %, and phosphorus source content is 270ppm weight %;
6) with the part enzymolysis product thin up in the step 3) to 1/10 of weight, obtain nutrient solution, nutrient solution is dropped into seeding tank; Be heated to 121 ℃ of sterilizations, keep after 30 minutes fast cooling to 36 ℃, insert aspergillus niger strain (black mold T01; Tianjin industrial microorganism institute; Equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of nutrient solution is a benchmark, inoculation 3 * 10
5Individual spore), be 4.0,0.4 volume at 36 ℃, original ph: carry out spawn culture under the aeration condition of volume minute; Measure through sampling sediments microscope inspection, acid test and pH the growth of black mold observed, after 25 hours, when pH 2.0, acidity 10g/L, bacterium ball size evenly, mycelia is sturdy when stretching out, stop to cultivate;
7) fermentation: 5) insert 6 in the substratum of gained) in cultivate aspergillus niger strain (black mold Co827, Tianjin industrial microorganism institute, equal aspergillus niger strain for this reason in the embodiment of the invention, inoculum size is: every liter of fermention medium is a benchmark, inoculation 3 * 10
5Individual spore), and the ventilation oxygen supply, the pH value is 5, tank pressure is about 0.06Mpa, air flow 0.3V/Vmin, under the condition of 37 ℃ of culture temperature, cultivate 58h after, as residual reducing sugar 9g/L, stop jar, terminal point acidity is 167g/L.
8) in the preparation of waste liquid: with 7) citric acid fermentation broth that obtains carries out press filtration to remove thalline, obtains fermentation clear liquid after the press filtration, under 75 ℃; Lime carbonate is slowly joined in the citric acid fermentation clear liquid (content of Hydrocerol A is 16.4 grams in every liter of citric acid fermentation clear liquid), and control pH value is 4.8 o'clock, stops to add lime carbonate; Reacted 60 minutes; After the layering of question response liquid, filter, the filtrating that obtains is in once and waste liquid.
Comparative Examples 1
This Comparative Examples is used to explain the preparation of the citric acid fermentation broth of prior art.
Method according to embodiment 5 is produced citric acid fermentation broth, and different is only to size mixing with former water (fresh water); The weight ratio of said starchy material and said adjusting slurry is 1: 3.2, and in addition, the fermentation termination judgement criteria in the step 7) does; After cultivating 68h; Residual reducing sugar is 0g/L, stops jar, and terminal point acidity is 141g/L.
From The above results, can see; Citric acid fermentation broth based in the Comparative Examples of the citric acid fermentation broth of the embodiment 1-5 of method of the present invention preparation and method preparation through prior art is compared, owing to replace water to allocate with waste liquid in using, can reduce the process water amount greatly; And in waste liquid can be used in the process of sizing mixing again; Therefore need not wait until complete fermentation, shorten the production cycle, improve production efficiency; Also reached the purpose of saving the energy, water reuse also helps environmental protection; And since in return and join the high problem of residual sugar that solved with waste liquid, therefore can improve the fermentation pol, finally obtain the high concentration of citric zymotic fluid.
Claims (9)
1. the preparation method of a citric acid fermentation broth is characterized in that, this method comprises:
The step of sizing mixing: adjusting slurry is mixed with starchy material, obtain farinaceous size; Said adjusting slurry is for containing the lemon aqueous acid; Said contain the lemon aqueous acid contain come from fermentation step obtain producing in the process that citric acid fermentation broth purifies in and waste liquid, the weight ratio of said starchy material and said adjusting slurry is 1: 1.86-3;
Enzymolysis step: under enzymatic hydrolysis condition, the farinaceous size that the step of sizing mixing is obtained mixes with enzyme and carries out enzymolysis, obtains enzymolysis product;
Fermentation step: under the condition that generates Hydrocerol A, the enzymolysis product that enzymolysis step is obtained ferments, and obtains citric acid fermentation broth; Fermentation termination is the residual reducing sugar content 1-10g/L of fermented liquid.
2. method according to claim 1, wherein, in the step of sizing mixing, contain the lemon aqueous acid and be come from in obtaining in the fermentation step producing in the process that citric acid fermentation broth purifies with waste liquid or should in the mixture of waste liquid and water.
3. method according to claim 2 wherein, is a benchmark with the volume that contains the lemon aqueous acid, said in waste liquid account for 40-60 volume %, said water accounts for 40-60 volume %.
4. according to claim 1,2 or 3 described methods, wherein, in the step of sizing mixing, in Citric acid monohydrate Food grade, said 1 liter of content that contains Hydrocerol A in the lemon aqueous acid is the 1-3 gram.
5. method according to claim 1, wherein, in the step of sizing mixing, the weight ratio of said starchy material and said adjusting slurry is 1: 2.13-2.7.
6. according to claim 1 or 5 described methods, wherein, in the step of sizing mixing, the pH value of said farinaceous size is 5.0-6.5.
7. method according to claim 1, wherein, in the enzymolysis step, said enzymatic hydrolysis condition comprises: hydrolysis temperature is 70-98 ℃, and enzymolysis time is 60-180 minute, and the pH of enzymolysis is 5.0-6.5.
8. method according to claim 1, wherein, in the fermentation step, fermentation termination is that the residual reducing sugar content of fermented liquid is 2-9g/L.
9. according to claim 1 or 8 described methods, wherein, in the fermentation step; Said fermentation condition comprises: the temperature of fermentation is 25-45 ℃, and the pH value is 1-7, and tank pressure is 0.05-0.1Mpa; Air flow is the 0.15-0.5 volume: volume minute, the time of fermentation is 45-75 hour.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102851330A (en) * | 2012-09-21 | 2013-01-02 | 中粮生物化学（安徽）股份有限公司 | Method for preparing citric acid through fermentation |
| CN105821083A (en) * | 2016-04-11 | 2016-08-03 | 马鞍山中粮生物化学有限公司 | Preparation method of citric acid |
| CN110885136A (en) * | 2018-09-07 | 2020-03-17 | 中粮生物化学（安徽）股份有限公司 | Treatment method of calcium citrate washing wastewater and preparation method of citric acid |
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| US4994609A (en) * | 1990-06-06 | 1991-02-19 | Innova S.A. | Production of citric acid |
| CN1300854A (en) * | 1999-12-18 | 2001-06-27 | 淄博华航药业有限公司 | Process for utilizing waste saccharide liquid in producing citric acid by calcium salt method and process for preparing citric acid |
| CN101948881A (en) * | 2010-08-10 | 2011-01-19 | 宜兴协联生物化学有限公司 | Method for producing citric acid by fermentation of mixed raw materials |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US4994609A (en) * | 1990-06-06 | 1991-02-19 | Innova S.A. | Production of citric acid |
| CN1300854A (en) * | 1999-12-18 | 2001-06-27 | 淄博华航药业有限公司 | Process for utilizing waste saccharide liquid in producing citric acid by calcium salt method and process for preparing citric acid |
| CN101948881A (en) * | 2010-08-10 | 2011-01-19 | 宜兴协联生物化学有限公司 | Method for producing citric acid by fermentation of mixed raw materials |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102851330A (en) * | 2012-09-21 | 2013-01-02 | 中粮生物化学（安徽）股份有限公司 | Method for preparing citric acid through fermentation |
| CN105821083A (en) * | 2016-04-11 | 2016-08-03 | 马鞍山中粮生物化学有限公司 | Preparation method of citric acid |
| CN110885136A (en) * | 2018-09-07 | 2020-03-17 | 中粮生物化学（安徽）股份有限公司 | Treatment method of calcium citrate washing wastewater and preparation method of citric acid |
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