CN105861575A - Citric acid fermentation method - Google Patents
Citric acid fermentation method Download PDFInfo
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- CN105861575A CN105861575A CN201610408924.4A CN201610408924A CN105861575A CN 105861575 A CN105861575 A CN 105861575A CN 201610408924 A CN201610408924 A CN 201610408924A CN 105861575 A CN105861575 A CN 105861575A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/44—Polycarboxylic acids
- C12P7/48—Tricarboxylic acids, e.g. citric acid
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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Abstract
The invention relates to a citric acid fermentation method. Sorghum and cassava are used as raw materials and subjected to smashing, liquidation after size mixing, and plate-frame pressure filtration, sorghum flour liquidation fluid and cassava flour liquidation fluid are added into a seed tank, a certain amount of inorganic nitrogen or/and organic nitrogen is also added, the rest of sorghum flour liquidation fluid and cassava flour liquidation fluid are introduced into a fermentation tank directly or after being subjected to plate-frame filtration, a certain amount of inorganic nitrogen or/and organic nitrogen is added, then disinfection and cooling are conducted, and finally seed tank culture solution is transferred into the fermentation tank so that fermentation can be started. By the adoption of the technical scheme, non-GMO materials are adopted, raw material source is wide, raw material risk and cost are reduced, the economic benefits of enterprises are further increased, and the industrial competitiveness of enterprises is improved.
Description
Technical field
A kind of method that the present invention relates to citric acid fermentation, particularly relates to one and ferments with Sorghum vulgare Pers. and Maninot esculenta crantz. batch mixing
The method producing citric acid.
Background technology
Citric acid, formal name used at school 2-hydroxy propane-1,2,3-tricarboxylic acids, molecular formula C6H8O7(anhydride), is a kind of
It is widely used in food, medicine and the organic acid of chemical field.Along with expanding economy, all trades and professions pair
The demand of citric acid will grow steadily, and the challenge faced by certain each enterprise also gets more and more with opportunity.Mainly
For food industry, pharmaceutical sector, chemical industry, and electronics, weaving, oil, leather, building,
The industrial circles such as photography, plastics, casting and pottery also there is the most wide application.
At present, the research and development fast development of genetically modified food, product variety and yield is also doubled and redoubled, transgenic
As a kind of emerging animal nutrition, it immature and uncertain so that the peace of genetically modified food
Full property becomes focus of concern, and Semen Maydis is then the key area in transgene agricultural product, and citric acid
Commercial production mainly uses and obtains through submerged fermentation for raw material with Semen Maydis, is therefore limited to Semen Maydis as main
, there is the problem that raw material the risk and cost is high in cereal crops and the risk of transgenic.At application number
CN201210347960.6 has described a kind of method utilizing Sorghum vulgare Pers. powder fermentation production of citric acid, can be significantly
Degree reduces the use of Semen Maydis, but it is also added into part Semen Maydis in seed tank culture base and fermentation tank culture medium
Powder liquefier.Therefore, it is still difficult to solve the dependence to maize raw material, it is impossible to accomplish that the most non-grain is non-
Transgenic fermenting raw materials obtains citric acid, and how to be capable of the fermentation of the most non-grain non-transgenic and obtain lemon
Lemon acid becomes one of prior art problem demanding prompt solution.
Summary of the invention
A kind of method that the present invention is to provide citric acid fermentation, uses non-grain non-transgenic fermenting raw materials to produce
Citric acid, has widened fermentation raw material source, effectively reduces raw material the risk and cost, solves and restrict lemon at present
Meng Suan manufacturing enterprise raw material sources and the high problem of cost of material, guarantee that domestic and international client is to non-turn of product simultaneously
The requirement of gene, the development for citric acid fermentation industry provides brand-new method and thinking.
The technical solution adopted in the present invention is as follows:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
(1) prepared by Sorghum vulgare Pers. and Maninot esculenta crantz. pulverizing respectively Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 10-100wt%,
Maninot esculenta crantz. accounts for 0-90wt%；
(2) being added water by Sorghum vulgare Pers. powder and size mixing, after sizing mixing, concentration is 15-35wt%, regulates pH5.5～6.5, and to
Wherein add the neutral protease of 10-25 enzyme unit of every gram of Sorghum vulgare Pers. powder, the cellulase of 0-20 enzyme unit and
The Thermostable α-Amylase of 5～25 enzyme units, 50-55 DEG C is incubated 30-120 minute；
Being added water by tapioca starch and size mixing, after sizing mixing, concentration is 15-35wt%, regulation pH5.5～6.5, and wherein
Add every gram of tapioca starch 5～the Thermostable α-Amylase of 15 enzyme units；
(3) Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing liquefy respectively and obtain Sorghum vulgare Pers. powder liquefier and tapioca starch
Liquefier, liquefier DE value is between 10～85%；
(4) step (3) will account for Sorghum vulgare Pers. powder liquefier or the tapioca starch liquid of final fermentation liquid cumulative volume 5-10%
Change liquid and squeeze in seed tank, add the 80-90 DEG C of hot water accounting for final fermentation liquid cumulative volume 4.97-9.995%, add
Enter to account for the inorganic nitrogen of final fermentating liquid volume 0.005-0.03% or/and organic nitrogen, at the 80-121 DEG C of 15-60 that sterilizes
Minute, it being cooled to 28-40 DEG C, access Aspergillus niger spores suspension and carry out seed liquor cultivation, its miospore accesses
Amount is 2.0-6.0*105Individual/mL, in incubation, speed of agitator is 50-200rpm, and tank pressure is
0.03-0.15MPa, ventilation is 1:0.05-0.4, cultivates 20-35h for 28-40 DEG C and i.e. may move in fermentation tank；
(5) by Sorghum vulgare Pers. powder liquefier remaining in step (3) and tapioca starch liquefier directly or through sheet frame mistake
Filter is driven in fermentation tank the 78.5-89.5% to fermentation liquid cumulative volume, adds afterwards and accounts for fermentation liquid cumulative volume
The inorganic nitrogen of 0.5-1.5%, or/and organic nitrogen, after 80-121 DEG C is sterilized 15-60 minute, is cooled to 35-40 DEG C；
(6) step (4) is cultivated ripe seed culture fluid and move in fermentation tank, control fermentation condition such as
Under: speed of agitator is 50-200rpm, and tank pressure is 0.03-0.15MPa, and ventilation ratio is 1:0.05-0.4, fermentation
Temperature 35-40 DEG C, fermentation, until fermentation ends, can extract the citric acid in fermentation liquid afterwards.
Wherein after Sorghum vulgare Pers. described in step (1) and Cassava crushing the transmitance of 60 mesh more than 70%；
Its pH is adjusted by adding pH adjusting agent after sizing mixing by Sorghum vulgare Pers. powder described in step (2) and tapioca starch
Joint, described pH adjusting agent is sodium hydroxide or soda or hydrochloric acid or sulphuric acid；
Step (2) cellulase consumption is 5-20 enzyme unit of every gram of Sorghum vulgare Pers. powder.
In step (3), liquefaction detailed process is: Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to not respectively
Same jet liquefaction device liquefies through twice consecutive spraying fluidification technique, the most double injection liquid
In metallization processes, an injection temperation controls at 95-105 DEG C, once maintains 1-8 in liquefaction laminar flow tank after injection
Hour, secondary injection temperature controls at 120-135 DEG C, maintains 1-8 after secondary injection in secondary liquefaction agitator tank
Individual hour, the final liquefier DE value that liquefies, between 10～85%, obtained Sorghum vulgare Pers. powder liquefier and Maninot esculenta crantz. respectively
Powder liquefier；
Inorganic nitrogen described in step (4) and (5) is selected from ammonium sulfate, ammonium nitrate, ammonium chloride, ammonium carbonate, bicarbonate
One or more in ammonium, diammonium phosphate, ammonia, ammonium citrate etc., described organic nitrogen selected from carbamide,
One or more in peanut cake powder, dried silkworm chrysalis meal, peptone etc.；
The extraction of the citric acid described in step (6) uses calcium salt method or chromatography；
The standard that in step (6), fermentation ends is: fermentation tank normally produces acid per hour less than 0.1%, reduction
Sugar is less than 1.0%.
The present invention is that, with the maximum difference of prior art, the mixing raw material employing Sorghum vulgare Pers. and Maninot esculenta crantz., and high
Fine strain of millet accounts for 10-100wt%, and Maninot esculenta crantz. accounts for 0-90wt%, uses complete non-grain non-transgenic crop to make first for this area
Citric acid is prepared, wherein Sorghum vulgare Pers. Main Nutrients: crude fat 3%, crude protein 8～11%, slightly fibre for fermenting raw materials
Dimension 2～3%, starch 65～70%, and the content that protein is in seed is usually 9～11%, wherein there are about 0.28%
Lysine, the methionine of 0.11%, the cystine of 0.18%, the tryptophan of 0.10%, the arginine of 0.37%,
The histidine of 0.24%, the leucine of 1.42%, the isoleucine of 0.56%, the phenylalanine of 0.48%, 0.30%
Threonine, the valine of 0.58%.In sorghum seed, the content of leucine and valine is slightly above Semen Maydis, and
Arginic content is slightly below again Semen Maydis.Other various amino acid whose content are roughly equal with Semen Maydis.Sorghum vulgare Pers. egg
White matter content is higher than Semen Maydis, but bad, lacking lysine and tryptophan, protein digestibility is low,
And owing to the intermolecular cross-linking of kafirin matter is more, and between protein and starch, there is the strongest knot
Closing key, cause enzyme to be difficult to enter decomposition, so that starch therein is difficult to be obtained by, and this is the most existing
Being difficult to one of the reason using Sorghum vulgare Pers. as raw material production citric acid in technology, the present inventor is the most sharp
This problem, wherein neutral egg is overcome by the mode adding neutral protease and Thermostable α-Amylase simultaneously
The addition of white enzyme can make protein bound starch be fully dissolved out effectively by Proteolytic enzyme, and control every gram of Sorghum vulgare Pers.
The consumption of the neutral protease of powder 10～25 enzyme units, can fully ensure that the hydrolysis of albumen, corresponding afterwards
Adding appropriate Thermostable α-Amylase can make the starch of abjection from protein-crosslinking fully liquefy, and improves starch
Yield；Protein after hydrolysis can provide enough nitrogen substances simultaneously, and this moieties can conduct
Organic nitrogen is applied in follow-up fermentation technology, thus reduces the addition of organic nitrogen, improves the utilization of raw material
Rate, reduces the overall cost of fermentation.
In Sorghum vulgare Pers., content of cellulose is higher, and has insoluble, is difficult to aborning be converted application, this
Bright, utilizing interpolation cellulase is available glucose by cellulose hydrolysis.And preferably control every gram
The consumption of the cellulase of 5-20 enzyme unit of Sorghum vulgare Pers. powder, with the neutral protease added, high temperature resistant alphalise starch
Enzyme has synergism, can fully ensure that the hydrolysis of cellulose, and the interpolation of cellulase can increase raw material
Utilization rate, and the quality of citric acid has been promoted.
Maninot esculenta crantz. is not the food of nutritive equilibrium, belongs to typical non-grain crop, major part in Maninot esculenta crantz. dry
Being starch, in fresh potato, starch is containing about 25%～30%, containing about 80% in potato is dry, and therefore can be as Fructus Citri Limoniae
One of raw material of fermentation of acid, but owing to tapioca root nitrogen content is few, need in follow-up sweat
Adding more external nitrogen source, thus may require that more organic nitrogen and inorganic nitrogen add, this is also existing
Technology utilize Maninot esculenta crantz. supplement the former of nitrogen source and carbon source as the cereal crops such as Semen Maydis must be added during fermentation raw material
Because of, and which also limits the Maninot esculenta crantz. development as raw material production citric acid technology.
Being based on this reason, inventor determines use Sorghum vulgare Pers. and the mixing raw material of Maninot esculenta crantz., and limits height
Fine strain of millet accounts for 10-100wt%, and Maninot esculenta crantz. accounts for 0-90wt%, under this weight ratio, uses Thermostable α-Amylase liquid
Change, it can be ensured that produce the carbon source needed for citric acid, the organic nitrogen of part is provided by Sorghum vulgare Pers. and Maninot esculenta crantz. simultaneously,
Thus reduce the consumption in external nitrogen source, it is ensured that the needs of sweat；And if tapioca starch consumption is too high, then
The additional nitrogen source problem as prior art, therefore inventor's effectively supplementing as Sorghum vulgare Pers. can be run into
Use.
Sorghum vulgare Pers. powder and tapioca starch after pulverizing add water and size mixing, and after sizing mixing, mass fraction is 15-35wt%.
The too low meeting of concentration of sizing mixing is greatly increased the difficulty of later stage liquefaction, and water consumption increase simultaneously can increase subsequent production energy
Consumption, and if excessive concentration of sizing mixing can cause system excessively thickness, be unfavorable for the liquefaction in later stage；Due to pH mistake
High or too low diastatic activity all can be made to be suppressed, it is unfavorable for the liquefaction in later stage, causes in compound
The starch of residual is more, causes the starch loss of part, causes the waste of resource, so inventor selects to lead to
Cross and add the pH that Sorghum vulgare Pers. powder sized mixing by chemicals sodium hydroxide, soda, hydrochloric acid, sulphuric acid etc. and be adjusted, adjust
PH5.5～6.5 after joint, under the conditions of this pH, the activity of Thermostable α-Amylase can be protected.
Sorghum vulgare Pers. slurry adds after sizing mixing as mentioned above neutral protease, is primarily due to kafirin matter
Intermolecular cross-linking more, and between protein and starch, there is the strongest associative key, directly by liquefaction difficulty
To be discharged completely by starch, can effectively destroy the combination of albumen and starch by adding protease, fully
Release starch, improves the yield of sugar.Neutral protein enzyme dosage is excessive can increase production cost, during consumption is too low
Property protease is little with the probability of the substrate protein association reaction in Sorghum vulgare Pers. powder, and the hydrolysis effect of albumen is the best, shadow
Ringing albumen to separate with starch, the yield of starch is on the low side；And if with cellulase with the use of, in utilization
Property protease, the synergism of cellulase and Thermostable α-Amylase so that albumen and the hydrolysis of cellulose
Effect is more preferable, is conducive to improving the yield of starch.
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry, through twice consecutive spraying fluidification, reduce the generation of non-fermented saccharide,
Injection temperation controls at 95-105 DEG C for the first time, once injection after liquefaction laminar flow tank in maintain 1-8 hour, two
Secondary injection temperation controls at 120-135 DEG C, maintains 1-8 hour after secondary injection in secondary liquefaction agitator tank,
After twice liquefaction, the DE value of liquefier all controls between 10～85%, obtains Sorghum vulgare Pers. powder liquefier and wood respectively
Potato starch liquefier, the present invention uses the mode of secondary injection, mainly by the high temp effect of secondary injection,
Can make not by the albuminous degeneration of enzymolysis, it is simple to filter, make starch granules expand dissolution simultaneously, improve amylase
Action effect, and then improve starch recoveries.
Take the above-mentioned Sorghum vulgare Pers. powder liquefier accounting for final fermentation liquid cumulative volume 5-10% or tapioca starch liquefier squeezes into kind
In sub-tank, add the 80-90 DEG C of hot water accounting for final fermentation liquid cumulative volume 4.97-9.995%, add and account for final sending out
Ferment liquid amasss the inorganic nitrogen of 0.005-0.03% or/and organic nitrogen, sterilizes 15-60 minute at 80-121 DEG C, cooling
To 28-40 DEG C, access Aspergillus niger spores suspension and carry out seed liquor cultivation；During Ben, adding hot water is to adjust
The carbon-nitrogen ratio of joint seed tank culture base is to meet the requirement of seed culture, if the too high strain of pol produced prosperous,
Move in fermentation tank and can enter decline phase too early and affect citric acid yield；Total sugar is spent low growth and is not strengthened
Real, immigration fermentation tank product acid is partially slow and fermentation period is the longest；Add nitrogen source and be to ensure that the need of growth
, the inorganic nitrogen of addition is ammonium sulfate, ammonium nitrate, ammonium chloride, diammonium phosphate, ammonia, ammonium citrate
One or more in Deng, organic nitrogen be the one in carbamide, peanut cake powder, dried silkworm chrysalis meal, peptone etc. or
Multiple.In this step, spore access amount is 2.0-6.0*105Individual/mL, the Aspergillus niger strain used is black
Aspergillosis Co827.
By remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier directly or in plate-and-frame filtration is driven into fermentation tank
To the 78.5-89.5% of fermentation liquid cumulative volume, add the inorganic nitrogen of the 0.5-1.5% accounting for fermentation liquid cumulative volume afterwards
Or/and organic nitrogen, after 80-121 DEG C is sterilized 15-60 minute, it is cooled to 35-40 DEG C；During Ben, for ensureing
Sweat growth need, addition inorganic nitrogen be ammonium sulfate, ammonium nitrate, ammonium chloride, diammonium phosphate,
One or more in carbamide, ammonia, ammonium citrate etc., organic nitrogen be carbamide, peanut cake powder, dried silkworm chrysalis meal,
One or more in peptone etc..
In foregoing, Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier can directly be driven in fermentation tank, it is possible to warp
Being driven into after plate-and-frame filtration in fermentation tank, prioritizing selection the latter, this is owing to filtering the reduction of after fermentation fluid viscosity,
Dissolved oxygen mass transfer is preferable, saves the energy, beneficially mass transfer and dissolved oxygen, promotes fermentation liquid quality；Even if but not
Through filtering, it is also possible to be applied in technical scheme.
Step (4) can be cultivated ripe seed culture fluid afterwards and move in above-mentioned fermentation tank, 35-40 DEG C leads to
Wind stirring fermentation is to end of a period of fermenting, and during this, owing to temperature is too low, growth accretion rate is slow,
Cause fermentation period long；Growth, metabolism are had a negative impact by the too high meeting of temperature, result in it
His heteroacid, metabolite, increase burden to follow-up extraction, so fermentation temperature controls at 35-40 DEG C.
When fermentation tank normally produces acid per hour less than 0.1%, and reducing sugar can terminate fermentation less than 1.0%.Finally
Fermentation liquid conventional calcium salt method can be used to extract citric acid therein.
The present invention produces citric acid with Sorghum vulgare Pers. and Maninot esculenta crantz. for fermenting raw materials, produces Fructus Citri Limoniae with current corn fermentation
Acid is compared, and both can reduce raw material risk, can ensure that again product non-transgenic.Specifications contrast feelings
Condition is as shown in the table:
Batch mixing fermentation technique of the present invention contrasts situation with original corn fermentation technical specification
The technology of the visible present invention can significantly reduce production cost and the product risks of enterprise, improves the warp of enterprise
Ji benefit and product competitiveness, widened the source of fermentation raw material, effectively reduces raw material the risk and cost, solves
Determine current restriction Citric Acid Production enterprise's raw material sources and Cost Problems, guaranteed product non-transgenic simultaneously,
Development for citric acid fermentation industry provides brand-new method and thinking.
Detailed description of the invention
Aspergillus niger strain employed in the present embodiment is aspergillus niger Co827.
Embodiment 1:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 90wt%, and Maninot esculenta crantz. accounts for
10wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 25wt%, and regulation pH is 5.8, and is added thereto to
The neutral protease of 15 enzyme units of every gram of Sorghum vulgare Pers. powder, the cellulase of 12 enzyme units and 10 enzyme units
Thermostable α-Amylase；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 25wt%, and regulation pH is 5.8, and is added thereto to
The Thermostable α-Amylase of 7 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 95 DEG C, once maintaining 2 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls at 125 DEG C,
Maintaining 2 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is 15.4%,
Obtain Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively；
Seed tank is squeezed into the 5% Sorghum vulgare Pers. powder liquefier accounting for final fermentation liquid cumulative volume, adds that to account for fermentation liquid overall
87 DEG C of hot water of long-pending 6.99%, add and account for the ammonium sulfate of fermentation liquid cumulative volume 0.01% as inorganic nitrogen-sourced, subsequently
Being warming up to 121 DEG C sterilize 30 minutes, access spore after being cooled to 37 DEG C, controlling Aspergillus niger spores concentration is
2.0*105Individual/mL, ventilation ratio is 1:0.1, and tank pressure is 0.15MPa, and speed of agitator is 80rpm, cultivates 28h
I.e. may move in fermentation tank；
Above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier are all squeezed in fermentation tank total to fermentation liquid
The 86.5% of volume, add account for fermentation liquid cumulative volume 0.5% ammonia and 0.25% ammonium sulfate as inorganic nitrogen
Source, it is organic nitrogen source that interpolation accounts for the cottonseed meal powder of fermentation liquid cumulative volume 0.75%；Opening stirring makes the rotating speed be
75rpm, is cooled to 36 DEG C after being warming up to 90 DEG C of sterilizations 30 minutes；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.25, and tank pressure is 0.05MPa, stirs
Mixing rotating speed is 100rpm, cultivates until fermentation ends under the conditions of 36 DEG C, and the standard that fermentation ends is: fermentation tank
The acid of normal product is per hour less than 0.1%, and reducing sugar is less than 1.0%.With this understanding, fermentation 58 is little altogether
Time, producing acid 17.45%, conversion ratio 99.15%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography to carry
Take the citric acid in fermentation liquid.
Embodiment 2:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 10wt%, and Maninot esculenta crantz. accounts for
90wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 23wt%, and regulation pH is 6.0, and is added thereto to
The neutral protease of 12 enzyme units of every gram of Sorghum vulgare Pers. powder and the high temperature resistant alphalise starch of 15 enzyme units；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 27wt%, and regulation pH is 6.0, and is added thereto to
The Thermostable α-Amylase of 10 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 97 DEG C, once maintaining 1.5 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls 128
DEG C, maintaining 3 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is
23%, obtain Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively；
Seed tank is squeezed into the 5% Sorghum vulgare Pers. powder liquefier accounting for final fermentation liquid cumulative volume, adds that to account for fermentation liquid overall
Long-pending 4.98%85 DEG C of hot water, add account for the ammonium sulfate of fermentation liquid cumulative volume 0.01% and 0.01% ammonia as nothing
Machine nitrogen source, then raises temperature to 90 DEG C and sterilizes 45 minutes, accesses spore, control aspergillus niger after being cooled to 28 DEG C
Spore concentration is 6.0*105Individual/mL, ventilation ratio is 1:0.12, and tank pressure is 0.10MPa, and speed of agitator is 80rpm,
Cultivate 28h i.e. to may move in fermentation tank；
Above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier are squeezed in fermentation tank to fermentation liquid cumulative volume
89%, add and account for 0.25% ammonium sulfate of fermentation liquid cumulative volume as inorganic nitrogen-sourced, add that to account for fermentation liquid overall
The carbamide of long-pending 0.75% is organic nitrogen source；Opening stirring makes rotating speed be 75rpm, is warming up to 90 DEG C and sterilizes 60 points
37 DEG C it are cooled to after clock；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.35, and tank pressure is 0.05MPa, stirs
Mixing rotating speed is 90rpm, cultivates until fermentation ends under the conditions of 37 DEG C, and the standard that fermentation ends is: fermentation tank is just
Often producing acid and be less than 0.1% per hour, reducing sugar is less than 1.0%.With this understanding, fermentation 64 hours altogether,
Producing acid 17.36%, conversion ratio 98.63%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography extraction to send out
Citric acid in ferment liquid.
Embodiment 3:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 65wt%, and Maninot esculenta crantz. accounts for
35wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 30wt%, and regulation pH is 5.5, and is added thereto to
The neutral protease of 17 enzyme units of every gram of Sorghum vulgare Pers. powder and the high temperature resistant alphalise starch of 12 enzyme units；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 30wt%, and regulation pH is 5.5, and is added thereto to
The Thermostable α-Amylase of 12 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 105 DEG C, once maintaining 3 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls at 120 DEG C,
Maintaining 2.5 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is 42%,
Obtain Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively；
Seed tank is squeezed into the 5% tapioca starch liquefier accounting for final fermentation liquid cumulative volume, adds that to account for fermentation liquid overall
Long-pending 4.97%90 DEG C of hot water, add the ammonium citrate work of the ammonium sulfate accounting for fermentation liquid cumulative volume 0.01% and 0.01%
For inorganic nitrogen-sourced, add and account for the cottonseed meal powder of fermentation liquid cumulative volume 0.01% as organic nitrogen source, then raise temperature to
Sterilizing 30 minutes for 121 DEG C, access spore after being cooled to 36.5 DEG C, controlling Aspergillus niger spores concentration is 4.0*105
Individual/mL, ventilation ratio is 1:0.17, and tank pressure is 0.12MPa, and speed of agitator is 90rpm, cultivates 30h
Move in fermentation tank；
Above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier are squeezed in fermentation tank to fermentation liquid cumulative volume
89%, add and account for 0.25% ammonium sulfate of fermentation liquid cumulative volume and as inorganic nitrogen-sourced, add that to account for fermentation liquid total
The cottonseed meal powder of volume 0.75% is organic nitrogen source；Opening stirring makes rotating speed be 75rpm, is warming up to 90 DEG C and disappears
Poison was cooled to 36 DEG C after 30 minutes；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.0.32, and tank pressure is 0.09MPa,
Speed of agitator is 85rpm, cultivates until fermentation ends under the conditions of 36 DEG C, and the standard that fermentation ends is: fermentation tank
The acid of normal product is per hour less than 0.1%, and reducing sugar is less than 1.0%.With this understanding, fermentation 62 is little altogether
Time, producing acid 17.41%, conversion ratio 98.36%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography to carry
Take the citric acid in fermentation liquid.
Embodiment 4:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 75wt%, and Maninot esculenta crantz. accounts for
25wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 27wt%, and regulation pH is 6.5, and is added thereto to
The neutral protease of 20 enzyme units of every gram of Sorghum vulgare Pers. powder and the high temperature resistant alphalise starch of 17 enzyme units；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 35wt%, and regulation pH is 6.5, and is added thereto to
The Thermostable α-Amylase of 15 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 98 DEG C, once maintaining 5 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls at 127 DEG C,
Maintaining 1.5 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is
30.8%, obtain Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively；
Seed tank is squeezed into the Sorghum vulgare Pers. powder liquefier of account for final fermentation liquid cumulative volume 3% and the tapioca starch liquid of 5%
Change liquid, add and account for 7.98%85 DEG C of hot water of fermentation liquid cumulative volume, add the sulphuric acid accounting for fermentation liquid cumulative volume 0.02%
Ammonium, as inorganic nitrogen-sourced, then raise temperature to 110 DEG C and sterilizes 35 minutes, accesses spore after being cooled to 36.5 DEG C,
Controlling Aspergillus niger spores concentration is 3.5*105Individual/mL, ventilation ratio is 1:0.23, and tank pressure is 0.10MPa, stirring
Rotating speed is 100rpm, cultivates 32h and i.e. may move in fermentation tank；
Above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier are squeezed in fermentation tank to fermentation liquid cumulative volume
83.5%, add that to account for the cottonseed meal powder of fermentation liquid cumulative volume 0.5% be organic nitrogen source；Open stirring and make rotating speed
For 85rpm, after being warming up to 110 DEG C of sterilizations 30 minutes, it is cooled to 36 DEG C；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.36, and tank pressure is 0.07MPa, stirs
Mixing rotating speed is 150rpm, cultivates until fermentation ends under the conditions of 36 DEG C, and the standard that fermentation ends is: fermentation tank
The acid of normal product is per hour less than 0.1%, and reducing sugar is less than 1.0%.With this understanding, fermentation 60 is little altogether
Time, producing acid 17.59%, conversion ratio 99.38%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography to carry
Take the citric acid in fermentation liquid.
Embodiment 5:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First Sorghum vulgare Pers. pulverizing is prepared Sorghum vulgare Pers. powder, and the transmitance of Sorghum vulgare Pers. powder 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 27wt%, and regulation pH is 5.6, and is added thereto to
The neutral protease of 20 enzyme units of every gram of Sorghum vulgare Pers. powder, the cellulase of 15 enzyme units and 20 enzyme units
Thermostable α-Amylase；
Sorghum vulgare Pers. slurry after sizing mixing liquefies through twice consecutive spraying fluidification technique, the most double
In injection liquefaction process, an injection temperation controls at 97 DEG C, once maintains 4.5 in liquefaction laminar flow tank after injection
Hour, secondary injection temperature controls at 133 DEG C, and after secondary injection, in secondary liquefaction agitator tank, maintenance 3 is little
Time, the final liquefier DE value that liquefies is 30.0%, it is thus achieved that Sorghum vulgare Pers. powder liquefier；
Seed tank is squeezed into the 6% Sorghum vulgare Pers. powder liquefier accounting for final fermentation liquid cumulative volume, adds that to account for fermentation liquid overall
86 DEG C of hot water of long-pending 6.99%, add and account for the ammonium sulfate of fermentation liquid cumulative volume 0.01% as inorganic nitrogen-sourced, subsequently
Being warming up to 121 DEG C sterilize 30 minutes, access spore after being cooled to 37 DEG C, controlling Aspergillus niger spores concentration is
3.0*105Individual/mL, ventilation ratio is 1:0.13, and tank pressure is 0.15MPa, and speed of agitator is 100rpm, cultivates
27h i.e. may move in fermentation tank；
Above-mentioned remaining Sorghum vulgare Pers. powder liquefier is all squeezed into 85.5% to fermentation liquid cumulative volume in fermentation tank, adds
Add and account for 0.50% ammonium sulfate of fermentation liquid cumulative volume as inorganic nitrogen-sourced, add and account for fermentation liquid cumulative volume 1.0%
Cottonseed meal powder is organic nitrogen source；Opening stirring makes rotating speed be 170rpm, after being warming up to 90 DEG C of sterilizations 30 minutes
It is cooled to 36 DEG C；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.37, and tank pressure is 0.1MPa, stirs
Mixing rotating speed is 170rpm, cultivates until fermentation ends under the conditions of 36 DEG C, and the standard that fermentation ends is: fermentation tank
The acid of normal product is per hour less than 0.1%, and reducing sugar is less than 1.0%.With this understanding, fermentation 62 is little altogether
Time, producing acid 17.32%, conversion ratio 98.97%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography to carry
Take the citric acid in fermentation liquid.
Embodiment 6:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 85wt%, and Maninot esculenta crantz. accounts for
15wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 35wt%, and regulation pH is 5.5, and is added thereto to
The neutral protease of 25 enzyme units of every gram of Sorghum vulgare Pers. powder, the cellulase of 12 enzyme units and 25 enzyme units
High temperature resistant alphalise starch；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 15wt%, and regulation pH is 5.5, and is added thereto to
Thermostable α-Amylase, the Thermostable α-Amylase of 5 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 97 DEG C, once maintaining 2.5 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls 120
DEG C, maintaining 3 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is
23%, obtain Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively；
Seed tank is squeezed into the Sorghum vulgare Pers. powder liquefier of account for final fermentation liquid cumulative volume 5% and adds that to account for fermentation liquid total
6.995%90 DEG C of hot water of volume, add ammonium sulfate and 0.0025% ammonia accounting for fermentation liquid cumulative volume 0.0025%
As inorganic nitrogen-sourced, then raise temperature to 121 DEG C and sterilize 30 minutes, after being cooled to 36.5 DEG C, access spore, control
Aspergillus niger spores concentration processed is 2.0*105Individual/mL, ventilation ratio is 1:0.0.35, and tank pressure is 0.05MPa, stirring
Rotating speed is 120rpm, cultivates 35h and i.e. may move in fermentation tank；
To squeeze into after above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier plate-and-frame filtration in fermentation tank to sending out
The 87.0% of ferment liquid cumulative volume, adds and accounts for the diammonium phosphate of fermentation liquid cumulative volume 0.25% and the chlorination of 0.25%
Ammonium is as inorganic nitrogen-sourced, and it is organic nitrogen source that interpolation accounts for the carbamide of fermentation liquid cumulative volume 0.5%；Open stirring to make to turn
Speed is 100rpm, is cooled to 38 DEG C after being warming up to 105 DEG C of sterilizations 40 minutes；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.12, and tank pressure is 0.12MPa, stirs
Mixing rotating speed is 100rpm, cultivates until fermentation ends under the conditions of 38 DEG C, and the standard that fermentation ends is: fermentation tank
The acid of normal product is per hour less than 0.1%, and reducing sugar is less than 1.0%.With this understanding, fermentation 56 is little altogether
Time, producing acid 17.85%, conversion ratio 102.00%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography
Extract the citric acid in fermentation liquid.
Embodiment 7:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 75wt%, and Maninot esculenta crantz. accounts for
25wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 15wt%, and regulation pH is 5.9, and is added thereto to
The neutral protease of 25 enzyme units of every gram of Sorghum vulgare Pers. powder and the Thermostable α-Amylase of 18 enzyme units；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 29wt%, and regulation pH is 5.9, and is added thereto to
Thermostable α-Amylase, the Thermostable α-Amylase of 12 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 100 DEG C, once maintaining 3.5 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls 120
DEG C, maintaining 4.5 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is
45%, obtain Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively；
Seed tank is squeezed into the Sorghum vulgare Pers. powder liquefier of account for final fermentation liquid cumulative volume 10% and adds that to account for fermentation liquid total
5.97%85 DEG C of hot water of volume, add ammonium sulfate and the 0.02% ammonium nitrate conduct accounting for fermentation liquid cumulative volume 0.01%
Inorganic nitrogen-sourced, then raise temperature to 121 DEG C and sterilize 30 minutes, access spore after being cooled to 36.5 DEG C, control black
Aspergillus spore concentration is 5.0*105Individual/mL, ventilation ratio is 1:0.0.35, and tank pressure is 0.05MPa, speed of agitator
For 120rpm, cultivate 20h and i.e. may move in fermentation tank；
To squeeze into after above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier plate-and-frame filtration in fermentation tank to sending out
The 82.5% of ferment liquid cumulative volume, adds and accounts for the diammonium phosphate of fermentation liquid cumulative volume 0.25% and the chlorination of 0.25%
Ammonium, as inorganic nitrogen-sourced, add and accounts for the cottonseed meal powder of fermentation liquid cumulative volume 0.5% and 0.5% Semen Brassicae campestris dregs of rice powder for having
Machine nitrogen source；Opening stirring makes rotating speed be 150rpm, is cooled to 40 DEG C after being warming up to 105 DEG C of sterilizations 20 minutes；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.20, and tank pressure is 0.06MPa, stirs
Mixing rotating speed is 90rpm, cultivates until fermentation ends under the conditions of 40 DEG C, and the standard that fermentation ends is: fermentation tank is just
Often producing acid and be less than 0.1% per hour, reducing sugar is less than 1.0%.With this understanding, fermentation 55 hours altogether,
Producing acid 17.76%, conversion ratio 100.91%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography to extract
Citric acid in fermentation liquid.
Embodiment 8:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 10wt%, and Maninot esculenta crantz. accounts for
90wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 23wt%, and regulation pH is 6.4, and is added thereto to
The neutral protease of 10 enzyme units of every gram of Sorghum vulgare Pers. powder and the cellulase of 10 enzyme units and 10 enzyme units
Thermostable α-Amylase；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 27wt%, and regulation pH is 6.4, and is added thereto to
The Thermostable α-Amylase of 10 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 97 DEG C, once maintaining 4 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls at 125 DEG C,
Maintaining 2 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is 19.4%,
Obtain Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively；
Seed tank is squeezed into the 5% Sorghum vulgare Pers. powder liquefier accounting for final fermentation liquid cumulative volume, adds that to account for fermentation liquid overall
Long-pending 4.98%86 DEG C of hot water, add the ammonium citrate work of the ammonium sulfate accounting for fermentation liquid cumulative volume 0.01% and 0.01%
For inorganic nitrogen-sourced, then raise temperature to 90 DEG C and sterilize 60 minutes, access spore after being cooled to 28 DEG C, control black
Aspergillus spore concentration is 6.0*105Individual/mL, ventilation ratio is 1:0.12, and tank pressure is 0.10MPa, and speed of agitator is
80rpm, cultivates 28h and i.e. may move in fermentation tank；
To squeeze into after above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier plate-and-frame filtration in fermentation tank to sending out
The 89% of ferment liquid cumulative volume, adds and accounts for 0.25% ammonium sulfate of fermentation liquid cumulative volume as inorganic nitrogen-sourced, and interpolation accounts for
The cottonseed meal powder of fermentation liquid cumulative volume 0.75% is organic nitrogen source；Opening stirring makes rotating speed be 75rpm, is warming up to
90 DEG C sterilization 60 minutes after be cooled to 37 DEG C；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.15, and tank pressure is 0.05MPa, stirs
Mixing rotating speed is 75rpm, cultivates until fermentation ends under the conditions of 37 DEG C, and the standard that fermentation ends is: fermentation tank is just
Often producing acid and be less than 0.1% per hour, reducing sugar is less than 1.0%.With this understanding, fermentation 63 hours altogether,
Producing acid 17.48%, conversion ratio 101.21%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography to extract
Citric acid in fermentation liquid.
Embodiment 9:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First prepared by Sorghum vulgare Pers. and Cassava crushing Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 65wt%, and Maninot esculenta crantz. accounts for
35wt%, the transmitance of Sorghum vulgare Pers. powder and tapioca starch 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 27wt%, and regulation pH is 6.0, and is added thereto to
The neutral protease of 15 enzyme units of every gram of Sorghum vulgare Pers. powder and the cellulase of 15 enzyme units and 20 enzyme units
Thermostable α-Amylase；
Tapioca starch is added water and sizes mixing so that it is concentration reaches 27wt%, and regulation pH is 6.0, and is added thereto to
The Thermostable α-Amylase of 15 enzyme units of every gram of tapioca starch；
Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction devices respectively through twice
Consecutive spraying fluidification technique liquefies, and in the most double injection liquefaction process, an injection temperation controls
At 97 DEG C, once maintaining 3 hours in liquefaction laminar flow tank after injection, secondary injection temperature controls at 130 DEG C,
Maintaining 3 hours in secondary liquefaction agitator tank after secondary injection, the final liquefier DE value that liquefies is 25%, point
Huo get Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier；
Seed tank is squeezed into the 5% Sorghum vulgare Pers. powder liquefier accounting for final fermentation liquid cumulative volume, adds that to account for fermentation liquid overall
Long-pending 4.98%85 DEG C of hot water, add and account for the ammonium sulfate of fermentation liquid cumulative volume 0.02% as inorganic nitrogen-sourced, rise subsequently
Temperature is sterilized 45 minutes to 100 DEG C, accesses spore after being cooled to 36.5 DEG C, controls Aspergillus niger spores concentration and is
3.5*105Individual/mL, ventilation ratio is 1:0.15, and tank pressure is 0.10MPa, and speed of agitator is 120rpm, cultivates
26h i.e. may move in fermentation tank；
To squeeze into after above-mentioned remaining Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier plate-and-frame filtration in fermentation tank to sending out
The 89% of ferment liquid cumulative volume, adds and accounts for 0.25% ammonium sulfate of fermentation liquid cumulative volume as inorganic nitrogen-sourced, and interpolation accounts for
The carbamide of fermentation liquid cumulative volume 0.25% and the cottonseed meal powder of 0.5% are organic nitrogen source；Opening stirring makes the rotating speed be
85rpm, is cooled to 36.5 DEG C after being warming up to 90 DEG C of sterilizations 30 minutes；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.20, and tank pressure is 0.075MPa,
Speed of agitator is 85rpm, cultivates until fermentation ends under the conditions of 36.5 DEG C, and the standard that fermentation ends is: fermentation
Tank normally produces acid per hour less than 0.1%, and reducing sugar is less than 1.0%.With this understanding, 63 are fermented altogether
Hour, producing acid 17.95%, conversion ratio 102.57%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatograph
Method extracts the citric acid in fermentation liquid.
Embodiment 10:
A kind of method of citric acid fermentation, it concretely comprises the following steps:
First Sorghum vulgare Pers. pulverizing is prepared Sorghum vulgare Pers. powder, and the transmitance of Sorghum vulgare Pers. powder 60 mesh is more than 70%；
Sorghum vulgare Pers. powder is added water and sizes mixing so that it is concentration reaches 25wt%, and regulation pH is 6.1, and is added thereto to
The neutral protease of 10 enzyme units of every gram of Sorghum vulgare Pers. powder and the cellulase of 8 enzyme units and 15 enzyme units
Thermostable α-Amylase；
Sorghum vulgare Pers. slurry after sizing mixing liquefies through twice consecutive spraying fluidification technique, the most double
In injection liquefaction process, an injection temperation controls at 95 DEG C, once maintains 2.5 in liquefaction laminar flow tank after injection
Hour, secondary injection temperature controls at 135 DEG C, maintains 4 hours after secondary injection in secondary liquefaction agitator tank,
The final liquefier DE value that liquefies is 42%, it is thus achieved that Sorghum vulgare Pers. powder liquefier；
Seed tank is squeezed into the 5% Sorghum vulgare Pers. powder liquefier accounting for final fermentation liquid cumulative volume, adds that to account for fermentation liquid overall
Long-pending 4.98%90 DEG C of hot water, add and account for the ammonium sulfate of fermentation liquid cumulative volume 0.02% as inorganic nitrogen-sourced, rise subsequently
Temperature is sterilized 30 minutes to 118 DEG C, accesses spore after being cooled to 32 DEG C, and controlling Aspergillus niger spores concentration is 4.0*105
Individual/mL, ventilation ratio is 1:0.18, and tank pressure is 0.12MPa, and speed of agitator is 110rpm, cultivates 30h
Move in fermentation tank；
Squeeze in fermentation tank to fermentation liquid cumulative volume after above-mentioned remaining Sorghum vulgare Pers. powder liquefier plate-and-frame filtration
89%, add and account for 0.25% ammonium sulfate of fermentation liquid cumulative volume as inorganic nitrogen-sourced, add and account for fermentation liquid cumulative volume
The peptone of 0.25% and the cottonseed meal powder of 0.5% are organic nitrogen source；Opening stirring makes rotating speed be 80rpm, heats up
It is cooled to 38 DEG C after sterilizing 30 minutes to 85 DEG C；
Moving in fermentation tank by cultivating ripe seed liquor, ventilation ratio is 1:0.20, and tank pressure is 0.095MPa,
Speed of agitator is 85rpm, cultivates until fermentation ends under the conditions of 38 DEG C, and the standard that fermentation ends is: fermentation tank
The acid of normal product is per hour less than 0.1%, and reducing sugar is less than 1.0%.With this understanding, fermentation 58 is little altogether
Time, producing acid 17.78%, conversion ratio 101.60%, fermentation liquid is delivered to extract workshop and is used calcium salt method or chromatography
Extract the citric acid in fermentation liquid.
Claims (8)
1. the method for a citric acid fermentation, it is characterised in that: it concretely comprises the following steps:
(1) prepared by Sorghum vulgare Pers. and Maninot esculenta crantz. pulverizing respectively Sorghum vulgare Pers. powder and tapioca starch, wherein Sorghum vulgare Pers. accounts for 10-100wt%,
Maninot esculenta crantz. accounts for 0-90wt%；
(2) being added water by Sorghum vulgare Pers. powder and size mixing, after sizing mixing, concentration is 15-35wt%, regulates pH5.5～6.5, and to
Wherein add the neutral protease of 10-25 enzyme unit of every gram of Sorghum vulgare Pers. powder, the cellulase of 0-20 enzyme unit and
The Thermostable α-Amylase of 5～25 enzyme units, 50-55 DEG C is incubated 30-120 minute；
Being added water by tapioca starch and size mixing, after sizing mixing, concentration is 15-35wt%, regulation pH5.5～6.5, and wherein
Add every gram of tapioca starch 5～the Thermostable α-Amylase of 15 enzyme units；
(3) Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing liquefy respectively and obtain Sorghum vulgare Pers. powder liquefier and tapioca starch
Liquefier, liquefier DE value is between 10～85%；
(4) step (3) will account for Sorghum vulgare Pers. powder liquefier or the tapioca starch liquid of final fermentation liquid cumulative volume 5-10%
Change liquid and squeeze in seed tank, add the 80-90 DEG C of hot water accounting for final fermentation liquid cumulative volume 4.97-9.995%, add
Enter to account for the inorganic nitrogen of final fermentating liquid volume 0.005-0.03% or/and organic nitrogen, at the 80-121 DEG C of 15-60 that sterilizes
Minute, it being cooled to 28-40 DEG C, access Aspergillus niger spores suspension and carry out seed liquor cultivation, its miospore accesses
Amount is 2.0-6.0*105Individual/mL, in incubation, speed of agitator is 50-200rpm, and tank pressure is
0.03-0.15MPa, ventilation is 1:0.05-0.4, cultivates 20-35h for 28-40 DEG C and i.e. may move in fermentation tank；
(5) by Sorghum vulgare Pers. powder liquefier remaining in step (3) and tapioca starch liquefier directly or through sheet frame mistake
Filter is driven in fermentation tank the 78.5-89.5% to fermentation liquid cumulative volume, adds afterwards and accounts for fermentation liquid cumulative volume
The inorganic nitrogen of 0.5-1.5%, or/and organic nitrogen, after 80-121 DEG C is sterilized 15-60 minute, is cooled to 35-40 DEG C；
(6) step (4) is cultivated ripe seed culture fluid and move in fermentation tank, control fermentation condition such as
Under: speed of agitator is 50-200rpm, and tank pressure is 0.03-0.15MPa, and ventilation ratio is 1:0.05-0.4, fermentation
Temperature 35-40 DEG C, fermentation, until fermentation ends, can extract the citric acid in fermentation liquid afterwards.
The method of citric acid fermentation the most according to claim 1, is characterized in that, step (1) is described
Sorghum vulgare Pers. and Cassava crushing after the transmitance of 60 mesh more than 70%.
The method of citric acid fermentation the most according to claim 1, is characterized in that, the Sorghum vulgare Pers. described in step (2)
Its pH is adjusted after sizing mixing by powder and tapioca starch by interpolation pH adjusting agent, and described pH adjusting agent is
Sodium hydroxide or soda or hydrochloric acid or sulphuric acid.
The method of citric acid fermentation the most according to claim 1, is characterized in that, cellulose in step (2)
Enzyme dosage is 5-20 enzyme unit of every gram of Sorghum vulgare Pers. powder.
The method of citric acid fermentation the most according to claim 1, is characterized in that, liquefaction tool in step (3)
Body process is: Sorghum vulgare Pers. slurry and Maninot esculenta crantz. slurry after sizing mixing are delivered to different jet liquefaction device warps respectively
Cross twice consecutive spraying fluidification technique to liquefy, the most double injection liquefaction process once sprays temperature
Degree controls at 95-105 DEG C, once maintains 1-8 hour in liquefaction laminar flow tank after injection, secondary injection temperature control
System, at 120-135 DEG C, maintains 1-8 hour after secondary injection in secondary liquefaction agitator tank, and liquefy final liquid
Change liquid DE value, between 10～85%, obtains Sorghum vulgare Pers. powder liquefier and tapioca starch liquefier respectively.
The method of citric acid fermentation the most according to claim 1, is characterized in that, step (4) and (5) are described
Inorganic nitrogen selected from ammonium sulfate, ammonium nitrate, ammonium chloride, ammonium carbonate, ammonium hydrogen carbonate, diammonium phosphate, ammonia
One or more in water, ammonium citrate etc., described organic nitrogen selected from carbamide, peanut cake powder, dried silkworm chrysalis meal,
One or more in peptone etc..
The method of citric acid fermentation the most according to claim 1, is characterized in that, described in step (6)
The extraction of citric acid uses calcium salt method or chromatography.
The method of citric acid fermentation the most according to claim 1, is characterized in that, fermentation in step (6)
The standard ended is: fermentation tank normally produces acid per hour less than 0.1%, and reducing sugar is less than 1.0%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108220349A (en) * | 2016-12-22 | 2018-06-29 | 中粮集团有限公司 | For efficient co-production of citric acid and Glucosamine fermentation medium and use its fermentation process |
| CN110819670A (en) * | 2019-11-30 | 2020-02-21 | 河南天冠车用生物燃料技术实验室有限公司 | Method for fermenting mixed raw materials by grouping liquefaction |
| CN112048526A (en) * | 2020-09-21 | 2020-12-08 | 山东柠檬生化有限公司 | High-conversion-rate production method of fermented citric acid |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698860A (en) * | 2009-10-30 | 2010-04-28 | 宜兴协联生物化学有限公司 | Method for producing citric acid by carrying out slag removal and fermentation on cassava |
| 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 |
| CN102864185A (en) * | 2012-09-19 | 2013-01-09 | 安徽丰原发酵技术工程研究有限公司 | Method for producing citric acid by fermenting sorghum powder |
| CN102876738A (en) * | 2012-09-14 | 2013-01-16 | 日照金禾博源生化有限公司 | Method for producing citric acid by using high-strength fermentation technology |
| CN103642855A (en) * | 2013-11-27 | 2014-03-19 | 日照金禾博源生化有限公司 | Method for protease fed-batch during citric acid fermentation process |
| CN104164458A (en) * | 2014-07-29 | 2014-11-26 | 中粮生物化学（安徽）股份有限公司 | Processing method of starch raw material and method for preparing citric acid |
| CN105586368A (en) * | 2016-03-16 | 2016-05-18 | 江苏国信协联能源有限公司 | Treatment method of sorghum grains and method for fermentation production of citric acid |
-
2016
- 2016-06-12 CN CN201610408924.4A patent/CN105861575B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101698860A (en) * | 2009-10-30 | 2010-04-28 | 宜兴协联生物化学有限公司 | Method for producing citric acid by carrying out slag removal and fermentation on cassava |
| 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 |
| CN102876738A (en) * | 2012-09-14 | 2013-01-16 | 日照金禾博源生化有限公司 | Method for producing citric acid by using high-strength fermentation technology |
| CN102864185A (en) * | 2012-09-19 | 2013-01-09 | 安徽丰原发酵技术工程研究有限公司 | Method for producing citric acid by fermenting sorghum powder |
| CN103642855A (en) * | 2013-11-27 | 2014-03-19 | 日照金禾博源生化有限公司 | Method for protease fed-batch during citric acid fermentation process |
| CN104164458A (en) * | 2014-07-29 | 2014-11-26 | 中粮生物化学（安徽）股份有限公司 | Processing method of starch raw material and method for preparing citric acid |
| CN105586368A (en) * | 2016-03-16 | 2016-05-18 | 江苏国信协联能源有限公司 | Treatment method of sorghum grains and method for fermentation production of citric acid |
Non-Patent Citations (1)
| Title |
|---|
| 肖克: "柠檬酸发酵工艺新进展", 《食品安全导刊》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108220349A (en) * | 2016-12-22 | 2018-06-29 | 中粮集团有限公司 | For efficient co-production of citric acid and Glucosamine fermentation medium and use its fermentation process |
| CN110819670A (en) * | 2019-11-30 | 2020-02-21 | 河南天冠车用生物燃料技术实验室有限公司 | Method for fermenting mixed raw materials by grouping liquefaction |
| CN112048526A (en) * | 2020-09-21 | 2020-12-08 | 山东柠檬生化有限公司 | High-conversion-rate production method of fermented citric acid |
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