CN108624630A - A method of shortening the citric acid fermentation period - Google Patents
A method of shortening the citric acid fermentation period Download PDFInfo
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Abstract
The present invention relates to citric acid fermentation fields, and in particular to a method of shortening the citric acid fermentation period.This method is that the nonionic surfactant of 1 2.5 ‰ w/v is added in fermentation tank culture link, improves aspergillus niger membrane passage, is conducive to nutriment and rapidly enters in cell body, accelerate cell metabolism in Citric Acid Fermentation；The mass transfer of oxygen is improved simultaneously, air consumption is reduced, fermentation period is made to greatly shorten, improves production efficiency, increases production capacity.
Description
Technical field
The present invention relates to citric acid fermentation fields, and in particular to a method of shortening the citric acid fermentation period.
Background technology
Citric acid (Citric acid) also known as citric acid, scientific name be 2- hydroxy propanes tricarboxylic acids, 2- hydroxy propanes -1,2,
3- tricarboxylic acids (2-hydroxypropane-1,2,3-tricarboxylic acid), molecular formula C6H8O7.It is water white transparency
Or translucent crystal or granular, particle sprills, it is odorless, though have strong tart flavour, it is pleasant, slightly after astringent taste, safe nothing
Poison is widely used as the acid of food and beverage.Citric acid and its esters can be widely applied to food, beverage, health products, doctor
The industry fields such as medicine, chemical industry, electronics, metallurgy, building, printing and dyeing, leather, washing.Since the increasingly extensive application of citric acid promotes
The demand of citric acid further increases, and citric acid consumption market gradually expands, and high efficiency production citric acid can greatly improve enterprise
Industry yield increases economic benefit.Fermentation period is long to limit fermentation yield, therefore how to shorten fermentation period, improves fermentation
Efficiency is of great significance to Citric Acid Production.
Accelerate the method for Citric Acid Production at present, it is most of by the way of changing culture medium composition or fermentation condition, but
But also production acid amount declines or cost improves while often shortening fermentation period；In addition it is exactly to research and develop novel bacterial, but this
The kind more difficult realization of method, and new strain there may be unstability equivalent risk.The present inventor is logical thus
Research is crossed, nonionic surfactant is used for Citric Acid Production for the first time, is acted on by research nonionic surfactant black
The characteristics of aspergillus, synthesis improvement fermentation process finally obtain a kind of method in low-cost high-efficiency shortening citric acid fermentation period.
Invention content
In view of the problems of the existing technology, the present invention provides a kind of method shortening the citric acid fermentation period, the party
Method main technical schemes are that the nonionic surfactant of ‰ w/v of 1-2.5 is added in fermentation tank culture link, coordinate corresponding hair
Ferment condition is fermented.Nonionic surfactant improves aspergillus niger permeability of cell membrane, be conducive to nutriment quickly into
Enter in cell body, accelerates cell metabolism；The mass transfer of oxygen is improved simultaneously, air consumption is reduced, fermentation period is made to greatly shorten, and is improved
Production efficiency increases production capacity.
Specific process step is as follows：
1) liquefied corn is taken, is put into seeding tank, nitrogen source is added, adds water to supply seeding tank zymotic fluid surplus, high temperature goes out
After bacterium, cooling down, activated aspergillus niger spore suspension is accessed, cultivates 25-29h, obtains seed liquor；
2) liquefied corn and corn liquefaction clear liquid input fermentation tank are taken, nitrogen source, high-temperature sterilization, cooling down is added；
3) seed liquor is transferred in fermentation tank by sterile transfer device, nonionic surfactant is added, in culture temperature
37 ± 0.5 DEG C of degree, ventilation quantity 3000-4500Nm3/ h, tank are pressed 0.05-0.1Mpa, are sent out under conditions of stir speed (S.S.) 80-100rpm
Ferment is extremely ended.
Wherein, the liquefied corn preparation process is：Crush maize adds water to size mixing, and Thermostable α-Amylase is added,
Injection liquefaction；
Further, 60 mesh screens are crossed after crush maize, make percent of pass 60% or more；Water temperature controls when water being added to size mixing
At 55-60 DEG C；The additive amount of alpha-amylase is 12-16 enzyme activity unit of every gram of corn flour；Injection liquefaction is that secondary high-temperature sprays
Penetrate liquefaction.
The corn liquefaction clear liquid is that liquefied corn is carried out plate compression, and what removal maize pulp obtained crosses filtering
Liquid；
The addition of liquefied corn described in step 1) is the 1-2% of ferment tank liquid total volume；The seed
Tank zymotic fluid nitrogen concentration is 0.25-0.5g/l；
Further, the high-temperature sterilization temperature described in step 1) is 110-120 DEG C, time 15-30min；The drop
Temperature is cooled to 32-36 DEG C.
The addition of liquefied corn described in step 2) is the 10-25% of ferment tank liquid total volume；The corn
The addition of liquefaction clear liquid is the 65-75% of ferment tank liquid total volume；A concentration of 5-8g/l of ferment tank liquid nitrogen source；
Further, the high-temperature sterilization temperature described in step 2) is 110-120 DEG C, time 15-30min；The drop
Temperature is cooled to 35-36 DEG C.
Nonionic surfactant described in step 3) be TWEEN Series, ethoxylated dodecyl alcohol (AEO series),
Any one in Triton X-100, addition are ‰ w/v of 1-2.5 of ferment tank liquid.
The seeding tank zymotic fluid is 1 with ferment tank liquid volume ratio:9.
Further, it be fermentation and acid is 0.1%w/v hereinafter, reduced sugar is 0.1%w/v simultaneously that fermentation, which terminates index,
Below.
Involved ferment tank liquid total volume refers to zymotic fluid total volume when fermentation tank starts fermentation in the present invention；
Other involved techniques are citric acid fermentation industry common process in the present invention, can refer to patent CN201210339843,
It is no longer repeated herein.
Corn employed in the present invention is drying corn, and drying corn can effectively prevent nutritional ingredient to be destroyed, crush
60 mesh screens are crossed afterwards and make it through rate 60% or more, this granule size can increase raw material heating surface area, and starch granules is made to inhale
Water expansion, gelatinization, improve heat treatment efficiency, shorten heat treatment time, while also more convenient on material transportation；Water is added to size mixing
Temperature control will not be gelatinized in 55-60 DEG C of certifiable cornstarch, prevent from causing transmission difficult；12-16 is added in every gram of corn flour
The Thermostable α-Amylase of each and every one enzyme activity unit, ensures liquefaction effect on the basis of reducing cost；Using secondary when liquefaction
High-temperature injection, temperature are respectively 95-96 DEG C and 128-130 DEG C, not only ensure that the function and effect of amylase, also equally make albumen
Qualitative change is flocculated, its calm and filtering is conducive to.
Seed tank culture base preparatory phase of the present invention, the addition of liquefied corn are the 1- of ferment tank liquid total volume
2%, nitrogen source content can be clearly controlled, ensures that nitrogen source is suitable, prevents excessive waste and the inhibition to aspergillus niger culture；110-
120 DEG C, the sterilization treatment of 30min both ensure that culture medium will not be destroyed because temperature is excessively high, while also ensure sterilization
Effect.
The addition of fermentation tank culture medium preparatory phase of the present invention, liquefied corn is ferment tank liquid total volume
The addition of 10-25%, corn liquefaction clear liquid are the 65-75% of ferment tank liquid total volume, can make ferment tank liquid nitrogen
Source amount is suitable, had both prevented from wasting caused by excess, while in turn ensuring the material base of thalline correlation synthesis and metabolism, and had made thalline
It can prevent from growing too fast reduction production acid, grew slow increase fermentation period with more suitable growth rate.
The present invention's was mainly characterized in that in the fermentation tank culture stage, adds nonionic surfactant, has in water
Non-ionizing feature, because nonionic surfactant is the amphiphilic knot for hydrophilic group with hydroxyl (- OH) or ehter bond (R-O-R ')
Structure molecule must contain multiple such groups since the hydrophily of hydroxyl and ehter bond is weak, in molecule and just show centainly
Hydrophily, making it in water and organic solvent has preferable dissolubility, and stability is high in the solution, is not easy by forceful electric power solution
The influence of matter inorganic salts and acid, alkali；There is no lethal effect to microorganisms such as bacteriums simultaneously, the cell membrane of eukaryocyte can be improved
Permeability, and it is highly stable, the actual requirement of prior art can be met with high pressure sterilization.Addition is ‰ w/v of 1-2.5, can be protected
Demonstrate,prove aspergillus niger activity.If being added excessively, the increase of cost and the cracking of cell, rupture can be caused, addition is very few, and expection is not achieved
Effect can not be such that cell leakage improves.In addition economically for, nonionic surfactant cheap market price will not
Cause the waste of cost.
In conclusion the method for the present invention handles the aspergillus niger permeability of cell membrane in Citric Acid Production, reach
The purpose for shortening fermentation period, makes fermentation period be greatly shortened in the prior art；Production efficiency is improved, comprehensive energy is reduced
Consumption, improves enterprise production level.
Specific implementation mode
The specific implementation mode of form by the following examples does further specifically the above of the present invention
It is bright, but the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to example below.It is all to be based on the above of the present invention
The technology realized all belongs to the scope of the present invention, and it is complete to be all made of conventional prior unless otherwise specified, in following embodiments
At.
Embodiment 1
1. taking the liquefied corn of the 1-2% of ferment tank liquid total volume, put into seeding tank, nitrogen source is added, uses water
Supply 50m3, make nitrogen concentration be 0.48g/L, sterilize under the conditions of 110-120 DEG C, 30min, cooling down to 32-36 DEG C,
Activated aspergillus niger spore suspension is accessed, about 28h is cultivated, obtains seed liquor, aspergillus niger spore amount reaches about 4 × 105A/
ml。
2. the liquefied corn of ferment tank liquid total volume 25% is taken to put into fermentation tank, ferment tank liquid total volume is taken
65% corn liquefaction clear liquid puts into fermentation tank, and nitrogen source is added, makes final nitrogen concentration for 6.3g/l, 110-120 DEG C, pressure
0.06Mpa, 30min sterilization treatment postcooling are cooled to 35-36 DEG C.
3. seed liquor is transferred in fermentation tank by sterile transfer device, nonionic surfactant Triton X- are added
100, addition is 1 ‰ w/v of ferment tank liquid, with 37 ± 0.5 DEG C of ventilation quantity 3000-4500Nm of cultivation temperature3/ h, tank pressure
Fermentation to fermentation and acid is 0.1%w/v hereinafter, reduced sugar is under conditions of 0.05-0.1Mpa, stir speed (S.S.) 80-100rpm
0.1%w/v is hereinafter, terminate fermentation.
The seeding tank zymotic fluid is 1 with ferment tank liquid volume ratio:9.
Wherein, liquefied corn preparation process is：Crush maize, excessively 60 mesh screens are percent of pass 60% or more, are added
55-60 DEG C of water is sized mixing, and Thermostable α-Amylase is added, and dosage is every gram of corn flour of 15 enzyme activity unit, and secondary high-temperature sprays liquid
Change, primary condensing temperature of spraying is 95-96 DEG C, sprays 128-130 DEG C of condensing temperature for the second time；Corn liquefaction clear liquid is by corn
Liquefier carries out plate compression, and what removal maize pulp obtained crosses cleaner liquid.
Fermentation period is set to foreshorten to 53h by the method, fermentation index is 3.49g/ (Lh).
Embodiment 2
1. taking the liquefied corn of ferment tank liquid total volume 1-2%, put into seeding tank, nitrogen source is added, is mended with water
Foot arrives 50m3, nitrogen concentration 0.49g/L sterilizes under the conditions of 110-120 DEG C, 30min, and cooling down is to 32-36 DEG C, access
Activated aspergillus niger spore suspension cultivates 28h, obtains seed liquor, and aspergillus niger spore amount reaches about 4 × 105A/ml.
2. the liquefied corn of ferment tank liquid total volume 25% is taken to put into fermentation tank, ferment tank liquid total volume is taken
65% corn liquefaction clear liquid input fermentation tank, addition nitrogen source, a concentration of 6.5g/L, then 110-120 DEG C, pressure 0.06Mpa,
30min sterilization treatment postcoolings are cooled to 35-36 DEG C.
3. seed liquor is transferred in fermentation tank by sterile transfer device, nonionic surfactant Triton X- are added
100, addition is 1.5 ‰ w/v of ferment tank liquid total volume, with 37 ± 0.5 DEG C of cultivation temperature, ventilation quantity 3000-
4500Nm3/ h, tank press 0.05-0.1Mpa, under conditions of stir speed (S.S.) 80-100rpm fermentation to fermentation and acid be 0.1%w/v with
Under, reduced sugar is 0.1%w/v hereinafter, terminating fermentation.
The seeding tank zymotic fluid is 1 with ferment tank liquid volume ratio:9.
Wherein, liquefied corn preparation process is：Crush maize, excessively 60 mesh screens are percent of pass 60% or more, are added
55-60 DEG C of water is sized mixing, and Thermostable α-Amylase is added, and dosage is every gram of corn flour of 15 enzyme activity unit, and secondary high-temperature sprays liquid
Change, primary condensing temperature of spraying is 95-96 DEG C, sprays 128-130 DEG C of condensing temperature for the second time；Corn liquefaction clear liquid is by corn
Liquefier carries out plate compression, and what removal maize pulp obtained crosses cleaner liquid.
Fermentation period is set to foreshorten to 51h by the method, fermentation index is 3.62g/ (Lh).
Embodiment 3
1. taking the liquefied corn of ferment tank liquid total volume 1-2%, put into seeding tank, nitrogen source is added, is mended with water
Foot arrives 50m3, nitrogen concentration 0.48g/L sterilizes under the conditions of 110-120 DEG C, 30min, and cooling down is to 32-36 DEG C, access
Activated aspergillus niger spore suspension cultivates about 28h, obtains seed liquor, and aspergillus niger spore amount reaches about 4 × 105A/ml.
2. the liquefied corn of ferment tank liquid total volume 25% is taken to put into fermentation tank, ferment tank liquid total volume is taken
65% corn liquefaction clear liquid input fermentation tank, addition nitrogen source, a concentration of 6.4g/L, 110-120 DEG C, pressure 0.06Mpa,
30min sterilization treatment postcoolings are cooled to 35-36 DEG C.
3. seed liquor is transferred in fermentation tank by sterile transfer device, non-ionic surfactant Tween -80 is added, spits
Warm -80 additions are 1 ‰ w/v of ferment tank liquid total volume, with 37 ± 0.5 DEG C of ventilation quantity 3000- of cultivation temperature
4500Nm3/ h, tank press 0.05-0.1Mpa, under conditions of stir speed (S.S.) 80-100rpm fermentation to fermentation and acid be 0.1%w/v with
Under, reduced sugar is 0.1%w/v hereinafter, terminating fermentation.
The seeding tank zymotic fluid is 1 with ferment tank liquid volume ratio:9.
Wherein, liquefied corn preparation process is：Crush maize, excessively 60 mesh screens are percent of pass 60% or more, are added
55-60 DEG C of water is sized mixing, and Thermostable α-Amylase is added, and dosage is every gram of corn flour of 15 enzyme activity unit, and secondary high-temperature sprays liquid
Change, primary condensing temperature of spraying is 95-96 DEG C, sprays 128-130 DEG C of condensing temperature for the second time；Corn liquefaction clear liquid is by corn
Liquefier carries out plate compression, and what removal maize pulp obtained crosses cleaner liquid.
Fermentation period is set to foreshorten to 52h by the method, fermentation index is 3.56g/ (Lh).
Embodiment 4
1. taking the liquefied corn of ferment tank liquid total volume 1-2%, put into seeding tank, nitrogen source is added, is mended with water
Foot arrives 50m3, nitrogen concentration 0.47g/L sterilizes under the conditions of 110-120 DEG C, 30min, and cooling down is to 32-36 DEG C, access
Activated aspergillus niger spore suspension cultivates 28h, obtains seed liquor, and aspergillus niger spore amount reaches about 4 × 105A/ml.
2. the liquefied corn of ferment tank liquid total volume 25% is taken to put into fermentation tank, ferment tank liquid total volume is taken
65% corn liquefaction clear liquid puts into fermentation tank, is added nitrogen source, concentration 6.5g/L, 110-120 DEG C, pressure 0.06Mpa, 30min
Sterilization treatment postcooling is cooled to 35-36 DEG C.
3. seed liquor is transferred in fermentation tank by sterile transfer device, nonionic surfactant AEO3, AEO3 is added
Addition is 1 ‰ w/v of ferment tank liquid total volume, with 37 ± 0.5 DEG C of cultivation temperature, ventilation quantity 3000-4500Nm3/ h,
Fermentation to fermentation and acid is 0.1%w/v hereinafter, reduced sugar under conditions of tank pressure 0.05-0.1Mpa, stir speed (S.S.) 80-100rpm
For 0.1%w/v hereinafter, terminating fermentation.
The seeding tank zymotic fluid is 1 with ferment tank liquid volume ratio:9.
Wherein, liquefied corn preparation process is：Crush maize, excessively 60 mesh screens are percent of pass 60% or more, are added
55-60 DEG C of water is sized mixing, and Thermostable α-Amylase is added, and dosage is every gram of corn flour of 15 enzyme activity unit, and secondary high-temperature sprays liquid
Change, primary condensing temperature of spraying is 95-96 DEG C, sprays 128-130 DEG C of condensing temperature for the second time；Corn liquefaction clear liquid is by corn
Liquefier carries out plate compression, and what removal maize pulp obtained crosses cleaner liquid.
Fermentation period is set to foreshorten to 53h by the method, fermentation index is 3.49g/ (Lh).
Comparative example
1. taking the liquefied corn of ferment tank liquid total volume 1-2%, put into seeding tank, nitrogen source is added, is mended with water
Foot arrives 50m3, nitrogen concentration 0.5g/L sterilizes under the conditions of 110-120 DEG C, 30min, and cooling down is to 32-36 DEG C, access
Activated aspergillus niger spore suspension cultivates 28h, obtains seed liquor, and aspergillus niger spore amount reaches about 4 × 105A/ml.
2. the liquefied corn of ferment tank liquid total volume 25% is taken to put into fermentation tank, ferment tank liquid total volume is taken
65% corn liquefaction clear liquid puts into fermentation tank, is added nitrogen source, concentration 6g/L, 110-120 DEG C, pressure 0.06Mpa, and 30min goes out
Bacterium processing postcooling is cooled to 35-36 DEG C.
3. seed liquor is transferred in fermentation tank by sterile transfer device, with 37 ± 0.5 DEG C of ventilation quantity 3000- of cultivation temperature
4500Nm3/ h, tank press 0.05-0.1Mpa, under conditions of stir speed (S.S.) 80-100rpm fermentation to fermentation and acid be 0.1%w/v with
Under, reduced sugar is 0.1%w/v hereinafter, terminating fermentation.
The seeding tank zymotic fluid is 1 with ferment tank liquid volume ratio:9.
Wherein, liquefied corn preparation process is：Crush maize, excessively 60 mesh screens are percent of pass 60% or more, are added
55-60 DEG C of water is sized mixing, and Thermostable α-Amylase is added, and dosage is every gram of corn flour of 15 enzyme activity unit, and secondary high-temperature sprays liquid
Change, primary condensing temperature of spraying is 95-96 DEG C, sprays 128-130 DEG C of condensing temperature for the second time；Corn liquefaction clear liquid is by corn
Liquefier carries out plate compression, and what removal maize pulp obtained crosses cleaner liquid.
Fermentation period is set to foreshorten to 60h by the method, fermentation index is 3.11g/ (Lh).
Implementation result compares
| Using technology | Fermentation index (g/L.h) | Period (h) |
| Comparative example | 3.11 | 60 |
| Embodiment 1 | 3.41 | 55 |
| Embodiment 2 | 3.55 | 54 |
| Embodiment 3 | 3.62 | 53 |
| Embodiment 4 | 3.52 | 55 |
It is compared from the implementation result of upper table as can be seen that comparing common process, the method for the invention is to citric acid fermentation
Either there is prodigious optimization from production acid amount or fermentation period, is said from another level, and reduce throwing to a certain degree
Enter cost, improve production efficiency, is very suitable for enterprise's promotion and implementation.
Claims (6)
1. a kind of method shortening the citric acid fermentation period, it is characterised in that：‰ w/v of 1-2.5 are added in fermentation tank culture link
Nonionic surfactant.
2. a kind of method shortening the citric acid fermentation period according to claim 1, it is characterised in that：The nonionic
Surfactant is any one in TWEEN Series, ethoxylated dodecyl alcohol, Triton X-100.
3. a kind of method shortening the citric acid fermentation period according to claim 1, it is characterised in that：The fermentation
Journey specific process step is as follows：
1) liquefied corn is taken, is put into seeding tank, nitrogen source is added, water is added to supply seeding tank zymotic fluid surplus, high-temperature sterilization, drop
After temperature is cooling, activated aspergillus niger spore suspension is accessed, cultivates 25-29h, obtains seed liquor；
2) liquefied corn and corn liquefaction clear liquid input fermentation tank are taken, nitrogen source, high-temperature sterilization, cooling down is added；
3) seed liquor is transferred in fermentation tank by sterile transfer device, nonionic surfactant is added, in cultivation temperature 37
± 0.5 DEG C, ventilation quantity 3000-4500Nm3/h, tank presses 0.05-0.1Mpa, ferment under conditions of stir speed (S.S.) 80-100rpm to
It ends.
4. a kind of method shortening the citric acid fermentation period according to claim 3, it is characterised in that：
The liquefied corn preparation process is：Crush maize adds water to size mixing, and Thermostable α-Amylase, injection liquefaction is added；
60 mesh screens are crossed after crush maize, make percent of pass 60% or more；Water temperature control is at 55-60 DEG C when water being added to size mixing；Alpha-amylase
Additive amount be 12-16 enzyme activity unit of every gram of corn flour；Injection liquefaction is secondary high-temperature injection liquefaction；
The corn liquefaction clear liquid is that liquefied corn is carried out plate compression, and what removal maize pulp obtained crosses cleaner liquid.
5. a kind of method shortening the citric acid fermentation period according to claim 3, it is characterised in that：Described in step 1)
The addition of liquefied corn is the 1-2% of ferment tank liquid total volume；The seeding tank zymotic fluid nitrogen concentration is
0.25-0.5g/l。
6. a kind of method shortening the citric acid fermentation period according to claim 3, it is characterised in that：Described in step 2)
The addition of liquefied corn is the 10-25% of ferment tank liquid total volume；The addition of the corn liquefaction clear liquid is hair
The 65-75% of fermenting pot liquid total volume；A concentration of 5-8g/l of ferment tank liquid nitrogen source.
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