CN107881204A - A kind of method of fermentation of Aspergillus niger production sodium gluconate - Google Patents
A kind of method of fermentation of Aspergillus niger production sodium gluconate Download PDFInfo
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- CN107881204A CN107881204A CN201711192191.6A CN201711192191A CN107881204A CN 107881204 A CN107881204 A CN 107881204A CN 201711192191 A CN201711192191 A CN 201711192191A CN 107881204 A CN107881204 A CN 107881204A
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Abstract
The present invention is a kind of method of fermentation of Aspergillus niger production sodium gluconate, it is characterised in that including preparing black-koji mould suspension；Nutritive salt one is added in using glucose as the seeding tank of raw material and is configured to seed culture fluid, access black-koji mould suspension carries out seed culture；Nutritive salt two is added in using glucose as the seeding tank of raw material and is configured to zymotic fluid, access seed liquor is fermented, mycelia is detected during the fermentation and adjusts ventilation and rotating speed, the control of aspergillus niger Hyphal length is set to terminate to ferment when the glucose content in fermentation tank is less than 3g/L in the range of 10~15pm；Zymotic fluid obtains sodium gluconate finished product by filtering, decolouring, crystallization, separation, drying and other steps.For technology of the method compared to existing fermentation of Aspergillus niger production sodium gluconate in the present invention, yellow tank, smelly tank phenomenon will not be produced, and fermentation duration can be shortened, improves the yield and quality of sodium gluconate.
Description
Technical field
The present invention relates to bioengineering field, and sodium gluconate is produced more specifically to a kind of fermentation of Aspergillus niger
Method.
Background technology
Sodium gluconate is a kind of good, nontoxic, the non-corrosive acylate of stability, be widely used in medicine, food,
Chemical industry, light industry etc., cement blending agent is used as such as in construction industry, certain amount sodium gluconate is added in cement, can be increased
Add the plasticity and intensity of concrete；For acid-base balance in human body can be adjusted in terms of medicine, make nerve recovery normal；Eating
Conduct industry can be used as food additives.Sodium gluconate is chemically pure reagent, non-corrosiveness, and constant mass, these feature energy
Ensure that it has reliable and repeated result in the application.
The production method of sodium gluconate mainly has electrolysis, catalytic oxidation, two enzymes method, microbe fermentation method etc. at present
Method.Wherein energy consumption is big in the industrial production for electrolysis, is difficult to control；There is during oxidation in process of production for catalytic oxidation
Between long, the problems such as side reaction is more, and catalyst is after certain number is recycled, and catalytic efficiency declines；Two enzymes method, which exists, to expand
Limitation is dissipated, and the cost of enzyme is higher；And microbe fermentation method mild condition, obvious be increasingly becoming of energy-conservation produce sodium gluconate
Main stream approach.
Gluconic acid and its salt are mainly fermented in fermenting and producing using black-koji mould, are added in fermentation tank big
Culture medium is measured, access seed liquor is fermented, control cell concentration, pH, temperature, fermentation tank rotating speed, dissolved oxygen concentration (DO), residual
The indexs such as sugar, concentration.But as the seed selection of excellent species can be resistant to higher concentration of substrate, the addition of mass propgation base
And the yield of sodium gluconate that accessory substance makes to obtain is not high, purity is relatively low, reaction time length, the phenomenon of the easy yellow smelly tank of tank.
Because certain cell concentration will be maintained per wholesale ferment, and it is that every a collection of fermentation of progress will cultivate a collection of strain, thalline training
Support and suppress with consumption of glucose in fermentation process, the purity of sodium gluconate, the speed of reaction, the generation of secondary metabolite,
It is all closely related with the state modulator in the ratio and course of reaction of initial nutritive salt in reaction.Sent out with the reaction of current this area
From the point of view of ferment present situation, the reaction time, ratio of the gluconic acid sodium content in original zymotic fluid was less than 32% generally more than 20 hours,
And zymotic fluid easily turns to be yellow because of controlling improper and secondary metabolite excessive, is smelly in fermentation process, so as to largely effect on
The yield and quality of sodium gluconate.
In view of this, a kind of method of fermentation of Aspergillus niger production sodium gluconate is worked out, utilizes rational nutritive salt ratio
Control in example and course of reaction to parameter, further shorten the time of fermenting and producing sodium gluconate, improve sodium gluconate
The problem of yield, lifting sodium gluconate quality are those skilled in the art's urgent need to resolve.
The content of the invention
It is an object of the invention to provide a kind of method of fermentation of Aspergillus niger production sodium gluconate, this method can improve Portugal
Grape sodium saccharate yield, lifting sodium gluconate quality, fully solve the problems, such as that prior art is present.
To solve the above problems, the technical solution adopted in the present invention is：
A kind of method of fermentation of Aspergillus niger production sodium gluconate, it is characterised in that comprise the following steps：
1) black-koji mould of a preservation is taken, scrapes upper strata spore, accesses in culture medium, is trained at a temperature of 28-32 DEG C
Support to covering with spore, after to be configured to bacteria suspension standby；The culture medium is improvement Martin's agar medium；
2) Glucose Liquid, nutritive salt one and defoamer are added in seeding tank and sterilized, the rear black-koji mould suspension that accesses enters
Row seed culture；The temperature for controlling seed liquor is 28-32 DEG C, pH 5-7, and dissolved oxygen is not less than 30%；The black-koji mould suspension
Access amount be 25ml/L；The concentration of the Glucose Liquid is 180-220g/L；The nutritive salt one is by 0.18~0.22g/L's
Potassium dihydrogen phosphate, 0.18~0.22g/L magnesium sulfate, 0.60~0.65g/L diammonium hydrogen phosphate and 0.20~0.24g/L bran
Skin forms；
3) Glucose Liquid, nutritive salt two and defoamer are added in fermentation tank and sterilized, the concentration of the Glucose Liquid is
180-220g/L；The nutritive salt two is by 0.078~0.080g/L potassium dihydrogen phosphate, 0.052~0.054g/L magnesium sulfate,
0.010~0.015g/L diammonium hydrogen phosphate and 0.040~0.042g/L urea composition；When measuring grape glycosyloxy in seeding tank
Change when enzyme activity reaches 280~320U/ml and access seed liquor, the temperature for controlling zymotic fluid is 28-32 DEG C, pH 5-7, and dissolved oxygen is
20%~30%, it is 10~15pm to control aspergillus niger Hyphal length, when the glucose content in fermentation tank is less than 3g/L
Terminate fermentation；
4) zymotic fluid of acquisition is subjected to filtering and impurity removing matter, decolouring, crystallization, crystal is separated with liquid, crystal is carried out
Dry, finally obtain sodium gluconate finished product.
Further, in step 2), the sterilizing methods is sterilize 20min under conditions of 115-120 DEG C, the defoaming
Agent is polyethers defoamer, and the addition of the defoamer is 0.25-0.30ml/L.Defoamer hydrophily in the present invention compared with
It is good, easily sprawled in foaming media, defoaming capacity is strong and after by high temperature sterilization, can recover emulsified state automatically, will not ferment
The separation of liquid reclaimed water oil, it is demulsified and influences to use.This defoamer have can environmental protection, efficiently, without the advantage such as color spot, hygroscopicity be low.
Further, in step 3), the access amount of the seed liquor is 10-15%.The quality of aspergillus niger seed liquor access
Fermentation results can be directly influenced with quantity, reached to a certain degree when measuring glucose oxidase in seeding tank and live in the present invention
During 280~320U/ml, the seed liquor for accessing 10-15% carries out fermenting and producing, and the thalline total amount and concentration of access disclosure satisfy that hair
The fermenting and producing requirement of fermentation tank, the seed for providing that sufficient metabolism is vigorous, vitality is strong can be produced for fermentation of Aspergillus niger, culture transferring is extremely
It can be mushroomed out after fermentation tank, lag phase is short, shortens the fermentation period in fermentation tank.It is relatively short when the seed culture time
When, thalline quantity is less, and slow-growing after inoculation, and product initially forms time retardation, extends fermentation period；When seed is trained
Support the time it is relatively long when, although bacterium amount is more, inoculation after thalline fail too early, cause production capacity to decline.When seed liquor
When access amount is smaller, during fermentation ends, there is very high residual sugar amount in zymotic fluid, cause the waste of resource；When connecing for seed liquor
Enter amount it is larger when, nutriment consumes rapidly, causes thalline early ageing further to influence production performance.
Further, in step 3), the sterilizing methods is sterilize 20min under conditions of 115-120 DEG C, the defoaming
Agent is polyethers defoamer, and the addition of the defoamer is 0.25-0.30ml/L.The defoamer is polyethers defoamer,
Defoamer hydrophily is preferable, is easily sprawled in foaming media, and defoaming capacity is strong and after by high temperature sterilization, can recover to emulsify shape automatically
State, it will not separate, be demulsified in fermented liquid reclaimed water oil and influence to use.This defoamer have environmental protection, efficiently, without color spot, moisture absorption
Property advantage, its suds ability such as low it is more superior than defoaming capacity, to suppress the generation of the foam of whole fermentation process.
Further, in step 3), the method for controlling aspergillus niger Hyphal length is to pass through Olympus high power video
Microscope is every two hours measured to the Hyphal length for obtaining sample, can reduce bacterium by increasing rotating speed and reducing oxygen-supply quantity
Filament length degree.By increasing rotating speed, aspergillus niger mycelia is sheared to a certain extent, polishing can reduce Hyphal length；It is black
The fermentation process of Aspergillus is the process of extremely oxygen consumption, can reduce Hyphal length to a certain extent by reducing oxygen-supply quantity,
Light transmittance in fermentation tank is further improved by reducing aspergillus niger Hyphal length.
Further, the color of zymotic fluid is kept to be rendered as white or milky all the time described in step 3) in fermentation tank；
The sample printing opacity in zymotic fluid after filtering mycelia is kept more than 75%.Keep the color of zymotic fluid prevents for white or milky
Yellow tank and smelly tank phenomenon are produced in fermentation process, it is the yield for maintaining sodium gluconate to keep sample transmittance.
Further, the discoloration method of sodium gluconate described in step 4) is activated carbon decolorizing, and its step is will fermentation
Liquid centrifuges, and takes supernatant, after by supernatant by being decolourized in the carbon post that is made up of granular activated carbon.Using granular activated carbon
Decoloration process, the decolouring quality of value in ferment of sodium gluconate liquid is improved, and then improve the decolouring shade and quality of product, primary crystallization
Reach food grade standard.
Further, in step 4), the method for the filtering is plate-frame filtering removal of impurity method；The method of the crystallization is
105-115 DEG C is concentrated into more than 70% content, crystallisation by cooling；The method of the separation is to be divided crystal and liquid using seperator
From；The method of the drying is fluidized bed drying.Wherein plate-frame filtering removal of impurities mass-energy reaches the purpose of aseptic filtration, solid to realize
Liquid separates；Above-mentioned method for crystallising can realize that the crystal grain of formation is uniform, and crystalline form is good, and crystal purity is high；Wherein seperator
Work area is big, and disposal ability is strong, can realize that crystal is rapidly separated with liquid, and good separating effect；Wherein use fluid bed
Crystal is dried, its rate of drying is high, and the thermal efficiency is high, is not likely to produce back-mixing, channel, glues phenomena such as wall, realizes sodium gluconate crystalline substance
The rapid draing of body.
The method of fermentation of Aspergillus niger production sodium gluconate of the present invention, at least has the advantage that：
1) method of the invention can completely inhibit caused yellow tank and smelly tank phenomenon in fermentation process；
2) method of the invention can control fermentation time in 18h or so, can be at least from the point of view of current fermentation present situation
Shorten 10% fermentation time；
3) method of the invention has the advantages of cost is low, simple and easy to do for enzyme process and other method；
4) method of the invention can improve sodium gluconate yield, lifting sodium gluconate quality.
Specific embodiment
In order that the technological means that the present invention realizes, improves feature, the purpose reached is more readily apparent, below in conjunction with implementation
Example carries out clear, complete description to the present invention.
The technical scheme in the embodiment of the present invention will be clearly and completely described below, it is clear that described implementation
Example only part of the embodiment of the present invention, rather than whole embodiments.It is common based on the embodiment in the present invention, this area
The every other embodiment that technical staff is obtained under the premise of creative work is not made, belong to the model that the present invention protects
Enclose.
Embodiment 1：
An aspergillus niger is taken, scrapes upper strata spore, is accessed in culture medium, bacterium is configured to after covering with spore in 30 DEG C of cultures
Suspension is standby；The culture medium is improvement Martin's agar medium；
Pure glucose 2.0kg, wheat bran 2.2g, potassium dihydrogen phosphate 2.0g, magnesium sulfate 2.0g, phosphorus are added in 15L culture tanks
The sour ammonium 6.2g of hydrogen two, defoamer 2.6ml, 10L is settled to, 115 DEG C sterilize 20 minutes, are then down to 36 DEG C, add 250ml bacterium and hang
Liquid, the temperature for controlling seed liquor are 30 DEG C；The pH for keeping seed liquor is 5.2；Dissolved oxygen (DO) >=30% is controlled, it is small to cultivate 20
When.
(one grade fermemtation tank) adds pure glucose 5.4kg, potassium dihydrogen phosphate 2.13g, magnesium sulfate in 50L fermentation tanks
1.43g, urea 1.11g, diammonium hydrogen phosphate 0.032g, defoamer 7.02ml, 27L is settled to, 115 DEG C sterilize 20 minutes, then
37 DEG C are down to, seed liquor is accessed, inoculum concentration 12%, with 350g/L NaOH adjustment control PH5.2, starts to ferment, control hair
The temperature of zymotic fluid is 30 DEG C, the control of aspergillus niger Hyphal length is kept dissolved oxygen 20%~30% in the range of 10~15pm,
Every 2 hours to bacterial type, light transmittance, be measured, terminate to ferment when the glucose content in fermentation tank is less than 3g/L.Portion
Divided data and control operation such as table 1：
The partial data of 1 embodiment of table 1 and control operation
3rd step, filtering：Plate-frame filtering removes mycelium.
4th step, decolourize：Zymotic fluid is decolourized using activated carbon.
5th step, crystallization：105-115 DEG C is concentrated into mass content more than 75%, crystallisation by cooling.
6th step, separation, drying：Seperator separates crystal with liquid, then fluidized bed drying, obtain sodium gluconate into
Product.Sodium gluconate purity is 98.6%, yield 95.4%, a length of 17.5h during fermentation, without yellow tank, smelly tank phenomenon, with background
The fermentation time (20h) mentioned in technology is compared, and shortens 12.5%.
Embodiment 2：
An aspergillus niger is taken, scrapes upper strata spore, is accessed in culture medium, bacterium is configured to after covering with spore in 30 DEG C of cultures
Suspension is standby；The culture medium is improvement Martin's agar medium；
Pure glucose 1.85kg, wheat bran 2.1g, potassium dihydrogen phosphate 1.9g, magnesium sulfate 1.9g, phosphorus are added in 15L culture tanks
The sour ammonium 6g of hydrogen two, defoamer 2.6ml, 10L is settled to, 115 DEG C sterilize 20 minutes, are then down to 36 DEG C, add 250ml bacterium and hang
Liquid, the temperature for controlling seed liquor are 30 DEG C；The pH for keeping seed liquor is 5.2, control dissolved oxygen (DO) >=30%, and culture 20 is small
When.
Second step, fermentation：(one grade fermemtation tank) adds pure glucose 5kg, potassium dihydrogen phosphate 2.11g, sulphur in 50L fermentation tanks
Sour magnesium 1.40g, urea 1.08g, diammonium hydrogen phosphate 0.28g, defoamer 7.02ml, 27L is settled to, 115 DEG C sterilize 20 minutes, so
After be down to 37 DEG C, access seed liquor, inoculum concentration 10%, with 350g/L NaOH adjustment control PH5.2, start to ferment, control
The temperature of zymotic fluid is 30 DEG C, makes the control of aspergillus niger Hyphal length in the range of 10~15pm, keep dissolved oxygen 20%~
30%, every 2 hours to bacterial type, index of refraction, be measured, terminate to send out when the glucose content in fermentation tank is less than 3g/L
Ferment.Partial data and control operation such as table 2：
The partial data of 2 embodiment of table 2 and control operation
3rd step, filtering：Plate-frame filtering removes mycelium.
4th step, decolourize：Zymotic fluid is decolourized using activated carbon.
5th step, crystallization：105-115 DEG C is concentrated into mass content more than 75%, crystallisation by cooling.
6th step, separation, drying：Seperator separates crystal with liquid, then fluidized bed drying, obtain sodium gluconate into
Product.Sodium gluconate purity is 98.4%, yield 94.8%, a length of 18h during fermentation, without yellow tank, smelly tank phenomenon.With background skill
The fermentation time (20h) mentioned in art is compared, and shortens 10%.
Embodiment 3：
An aspergillus niger is taken, scrapes upper strata spore, is accessed in culture medium, bacterium is configured to after covering with spore in 30 DEG C of cultures
Suspension is standby；The culture medium is improvement Martin's agar medium；
Pure glucose 2.15kg, wheat bran 2.4g, potassium dihydrogen phosphate 2.2g, magnesium sulfate 2.2g, phosphorus are added in 15L culture tanks
The sour ammonium 6.4g of hydrogen two, defoamer 2.6ml, 10L is settled to, 115 DEG C sterilize 20 minutes, are then down to 36 DEG C, add 250ml bacterium and hang
Liquid, the temperature for controlling seed liquor are 30 DEG C；The pH for keeping seed liquor is 5.2, control dissolved oxygen (DO) >=30%, and culture 20 is small
When.
Second step, fermentation：In 50L fermentation tanks (one grade fermemtation tank) add pure glucose 5.8kg, potassium dihydrogen phosphate 2.06g,
Magnesium sulfate 1.46g, urea 1.13g, diammonium hydrogen phosphate 0.38g, defoamer 7.02ml, 27L is settled to, 115 DEG C sterilize 20 minutes,
Then 37 DEG C are down to, seed liquor is accessed, inoculum concentration 14%, with 350g/L NaOH adjustment control PH5.2, starts to ferment, is controlled
The temperature of zymotic fluid processed is 30 DEG C, makes the control of aspergillus niger Hyphal length in the range of 10~15pm, keep dissolved oxygen 20%~
30%, every 2 hours to bacterial type, index of refraction, be measured, terminate to send out when the glucose content in fermentation tank is less than 3g/L
Ferment.
Partial data and control operation such as table 3：
The partial data of 3 embodiment of table 3 and control operation
3rd step, filtering：Plate-frame filtering removes mycelium.
4th step, decolourize：Zymotic fluid is decolourized using activated carbon.
5th step, crystallization：105-115 DEG C is concentrated into mass content more than 75%, crystallisation by cooling.
6th step, separation, drying：Seperator separates crystal with liquid, then fluidized bed drying, obtain sodium gluconate into
Product.Sodium gluconate purity is 98.5%, yield 95.1%, a length of 18h during fermentation, without yellow tank, smelly tank phenomenon.With background skill
The fermentation time (20h) mentioned in art is compared, and shortens 10%.
Other parts in the present embodiment it is identical with embodiment 1 with regard to not one by one repeat.
The present invention is not limited to above-mentioned preferred embodiment, and anyone should learn that what is made under the enlightenment of the present invention
Structure change, it is every with it is of the invention have it is same or similar as technical scheme, belong to protection scope of the present invention.
Claims (8)
- A kind of 1. method of fermentation of Aspergillus niger production sodium gluconate, it is characterised in that comprise the following steps：1) take one preservation black-koji mould, scrape upper strata spore, access culture medium in, cultivated at a temperature of 28-32 DEG C to Cover with spore, after to be configured to bacteria suspension standby；The culture medium is improvement Martin's agar medium；2) Glucose Liquid, nutritive salt one and defoamer are added in seeding tank and sterilized, the rear black-koji mould suspension that accesses is planted Son culture；The temperature for controlling seed liquor is 28-32 DEG C, pH 5-7, and dissolved oxygen is not less than 30%；The black-koji mould suspension connects It is 25ml/L to enter amount；The concentration of the Glucose Liquid is 180-220g/L；The nutritive salt one by 0.18~0.22g/L phosphoric acid Potassium dihydrogen, 0.18~0.22g/L magnesium sulfate, 0.60~0.65g/L diammonium hydrogen phosphate and 0.20~0.24g/L wheat bran group Into；3) Glucose Liquid, nutritive salt two and defoamer are added in fermentation tank and sterilized, the concentration of the Glucose Liquid is 180- 220g/L；The nutritive salt two is by 0.078~0.080g/L potassium dihydrogen phosphate, 0.052~0.054g/L magnesium sulfate, 0.010~0.015g/L diammonium hydrogen phosphate and 0.040~0.042g/L urea composition；When measuring grape glycosyloxy in seeding tank Change when enzyme activity reaches 280~320U/ml and access seed liquor, the temperature for controlling zymotic fluid is 28-32 DEG C, pH 5-7, and dissolved oxygen is 20%~30%, it is 10~15pm to control aspergillus niger Hyphal length, when the glucose content in fermentation tank is less than 3g/L Terminate fermentation；4) zymotic fluid of acquisition is subjected to filtering and impurity removing matter, decolouring, crystallization, crystal is separated with liquid, crystal is dried, Finally obtain sodium gluconate finished product.
- A kind of 2. method of fermentation of Aspergillus niger production sodium gluconate according to claim 1, it is characterised in that step 2) In, the sterilizing methods are the 20min that sterilized under conditions of 115-120 DEG C, and the defoamer is polyethers defoamer, described to disappear The addition of infusion is 0.25-0.30ml/L.
- A kind of 3. method of fermentation of Aspergillus niger production sodium gluconate according to claim 1, it is characterised in that step 3) In, the access amount 10-15% of the seed liquor.
- A kind of 4. method of fermentation of Aspergillus niger production sodium gluconate according to claim 1, it is characterised in that step 3) In, the sterilizing methods are the 20min that sterilized under conditions of 115-120 DEG C, and the defoamer is polyethers defoamer, described to disappear The addition of infusion is 0.25-0.30ml/L.
- A kind of 5. method of fermentation of Aspergillus niger production sodium gluconate according to claim 1, it is characterised in that step 3) In, it is described control aspergillus niger Hyphal length method be, by Olympus high power videomicroscopy every two hours to obtaining sample The Hyphal length of product is measured, and can reduce Hyphal length by increasing rotating speed and reducing oxygen-supply quantity.
- A kind of 6. method of fermentation of Aspergillus niger production sodium gluconate according to claim 1, it is characterised in that step 3) In, keep the color of zymotic fluid to be rendered as white or milky all the time in the fermentation tank；Keep after filtering mycelia in zymotic fluid Sample printing opacity more than 75%.
- A kind of 7. method of fermentation of Aspergillus niger production sodium gluconate according to claim 1, it is characterised in that step 4) In, the discoloration method of the sodium gluconate is activated carbon decolorizing, and its step takes supernatant for zymotic fluid is centrifuged, after will be upper Clear liquid in the carbon post that is made up of granular activated carbon by being decolourized.
- A kind of 8. method of fermentation of Aspergillus niger production sodium gluconate according to claim 1, it is characterised in that step 4) In, the method for the filtering is plate-frame filtering removal of impurity method；The method of the crystallization is concentrated into more than 70% for 105-115 DEG C and contained Amount, crystallisation by cooling；The method of the separation is to be separated crystal with liquid using seperator；The method of the drying is fluid bed Dry.
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| CN111334538A (en) * | 2020-03-20 | 2020-06-26 | 鄂州职业大学 | Method for producing gluconic acid by strengthening penicillium funiculosum fermentation glucose |
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