CN106566852A - Sodium glutamate separation and purification method - Google Patents
Sodium glutamate separation and purification method Download PDFInfo
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- CN106566852A CN106566852A CN201610789756.8A CN201610789756A CN106566852A CN 106566852 A CN106566852 A CN 106566852A CN 201610789756 A CN201610789756 A CN 201610789756A CN 106566852 A CN106566852 A CN 106566852A
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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
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/14—Glutamic acid; Glutamine
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- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/14—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof
- C07C227/18—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton from compounds containing already amino and carboxyl groups or derivatives thereof by reactions involving amino or carboxyl groups, e.g. hydrolysis of esters or amides, by formation of halides, salts or esters
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/38—Separation; Purification; Stabilisation; Use of additives
- C07C227/40—Separation; Purification
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Abstract
The invention discloses a sodium glutamate separation and purification method. The sodium glutamate separation and purification method comprises the following specific steps: inoculating corynebacterium glutamicum into a bouillon culture medium, carrying out culturing, uniformly coating the bottom of a culture dish with the obtained bacterial suspension, carrying out microwave processing, inoculating the bacterial suspension obtained in the previous step into a seed medium, carrying out culturing, inoculating the thallus obtained in the previous step into a fermentation medium according to the 5% of inoculum size, and carrying out fermentation; adding urea and sodium carbonate into the glutamic acid fermentation liquor, carrying out filtering and sterilizing by adopting a micro-filtration membrane, collecting the sterilized liquid, adding the sterilized liquid into a decoloring pump, adding activated carbon into the sterilized liquid, and collecting discolored liquid; and finally, adding the discolored liquid into a concentrating pan, and carrying out concentrating treatment, thus obtaining sodium glutamate coarse crystal. The invention provides the novel process for producing sodium glutamate, microwave mutation breeding is carried out on the corynebacterium glutamicum, microwave as electromagnetic wave can stimulate polar molecules such as protein and nucleotide to rapidly shake, so that the DNA structure changes, thus heritable variation occurs, and therefore, the acid production capacity of bacterial strains is greatly improved.
Description
Technical field
The invention belongs to glutamate production technical field, more particularly to a kind of sodium glutamate process for separation and purification.
Background technology
Sodium glutamate, the sodium of chemical name alpha-amido 1,3-propanedicarboxylic acid one is a kind of salt formed by sodium ion and glutamate ion.
Its Glutamic Acid is a kind of aminoacid, and sodium is a kind of metallic element.The main component of the flavouring agent monosodium glutamate commonly used in life is just
It is sodium glutamate.Fructus Lycopersici esculenti, bean product, yeast extract, some sharp cheese of fermentation, and fermentation or aminosal are produced
In the seasoning effect that product to be brought, the presence of glutamic acid is partly due to.
Glutamic acid is a kind of universal aminoacid：The self-produced glutamic acid of human body, it is mainly present in rich in egg with complex status
In the food of white matter, such as mushroom, Thallus Laminariae (Thallus Eckloniae), Fructus Lycopersici esculenti, nut, beans, meat, and most of milk product.In the food of part
Glutamic acid is with free form presence；And only this free-form glutamate, Glu can strengthen the delicate flavour of food.Fructus Lycopersici esculenti,
The bean product of fermentation, yeast extract, some sharp cheese, and fermentation or aminosal product (such as soy sauce or bean sauce) institute
In the seasoning effect that can be brought, the presence of glutamic acid is partly due to.
The content of the invention
It is an object of the invention to provide a kind of sodium glutamate process for separation and purification, by carrying out to corynebacterium glutamicum
Microwave irradiation breeding, substantially increases the acid producing ability of bacterial strain.
The present invention is achieved by the following technical solutions：
A kind of sodium glutamate process for separation and purification, comprises the following specific steps that：
S1, by the corynebacterium glutamicum of slant culture to logarithmic (log) phase be inoculated in broth bouillon cultivate；
Supernatant is abandoned in culture medium centrifugation, the thalline for obtaining is washed 2-3 time with pH7.0 phosphate buffer solutions, by thalline
It is dissolved in pH7.0 phosphate buffer solutions and prepares bacteria suspension；
S2, obtained bacteria suspension 3-5mL in S1 is spread evenly across into culture dish bottom carries out microwave treatment, wherein microwave work(
Rate is 600-800W, and pulse frequency is 2450MHz, and microwave treatment time is 45-60s；
Thalline after microwave treatment is scattered in normal saline and configures bacteria suspension；
S3, the bacterial suspension inoculation obtained in S2 is cultivated in seed culture medium；
S4, by S3 cultivate after thalline with 5% inoculum concentration be inoculated in fermentation medium ferment, fermentation temperature is 35
DEG C, fermentation time 2-3h obtains glutami acid fermentation liquor；
Mass fraction is added to be 25% carbamide and sodium carbonate, carbamide and sodium carbonate in S5, the glutami acid fermentation liquor in S4
Mass ratio be 1:1, obtain sodium glutamate fermentation liquid；
S6, the sodium glutamate fermentation liquid for obtaining in S5 is degerming by micro-filtrate membrane filtration, collection bacteria-removing liquid, wherein mocromembrane
For 0.2 μm of film, microfiltration temperature is 30 DEG C, and operation pressure reduction is 0.22MPa；
S7, by the bacteria-removing liquid collected in S4 add decolouring pump in, in bacteria-removing liquid add mass fraction be 1% activated carbon,
Decolouring 20min；
By the bacteria-removing liquid collected after centrifugation destaining solution containing activated carbon；
S8, by the destaining solution obtained in S5 add concentration pan in, carry out concentration in 65-80 DEG C, obtain sodium glutamate
Coarse crystal.
Further, cultivation temperature is 37 DEG C in the S1, and incubation time is 14-16h, and culture rotating speed is 220r/min.
Further, centrifugal rotational speed is 3200-3700r/min in the S1, and centrifugation time is 10min.
Further, cell concentration is 1 × 10 in bacteria suspension described in S18Individual/mL, thalline is dense in bacteria suspension described in S2
Spend for 1 × 108Individual/mL.
Further, cultivation temperature is 32 DEG C in S3, and culture rotating speed is 120-140r/min, and incubation time is 24h.
Further, seed culture based component described in S3 is：Glucose 25-30g/L, K2HPO40.2-0.3g/L, urine
Plain 5-6g/L, Mg2SO41.2-1.4g/L, Semen Maydis pulp 35-40g/L, with 0.1mol/L NaOH pH to 7.0-7.2 is adjusted.
Further, fermentation medium components are described in S4：Glucose 160-180g/L, K2HPO4 2.3-3.5g/L、
Mg2SO41.2g/L, Semen Maydis pulp 12-15g/L, methionine 0.6-0.9g/L, VB1 0.05-0.1mg/L。
Further, centrifugation rate described in S6 is 1700-2100r/min, and centrifugation time is 2min.
Further, centrifugation rate described in S7 is 3000r/min, and centrifugation time is 10min.
The invention has the advantages that：
The invention provides a kind of sodium glutamate production new technique, is educated by carrying out microwave irradiation to corynebacterium glutamicum
Kind, microwave, can be with stimulating protein and nucleotide isopolarity molecule fast vibration used as a kind of electromagnetic wave so that DNA structure is sent out
Changing, so as to hereditary variation occur, substantially increases the acid producing ability of bacterial strain.
Certainly, the arbitrary product for implementing the present invention it is not absolutely required to while reaching all the above advantage.
Specific embodiment
Technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is only
The a part of embodiment of the present invention, rather than the embodiment of whole.Based on the embodiment in the present invention, those of ordinary skill in the art
The all other embodiment obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
Embodiment 1
S1, the corynebacterium glutamicum of slant culture to logarithmic (log) phase is inoculated in broth bouillon in 37 DEG C of shaking baths
Middle culture 14h, shaking speed is 220r/min；
Culture medium is abandoned into supernatant with 3200r/min centrifugation 10min, by the thalline for obtaining pH7.0 phosphate buffer solutions
Thalline is dissolved in pH7.0 phosphate buffer solutions after washing 2 times prepares bacteria suspension, wherein cell concentration is 1 × 108Individual/mL；
S2, obtained bacteria suspension 3mL in S1 is spread evenly across into culture dish bottom carries out microwave treatment, wherein culture dish is straight
Footpath is 7cm, and microwave power is 600W, and pulse frequency is 2450MHz, and microwave treatment time is 45s；
Thalline after microwave treatment is scattered in normal saline and configures bacteria suspension, cell concentration be cell concentration be 1 ×
108Individual/mL；
S3, by the bacterial suspension inoculation obtained in S2 in seed culture medium, in 32 DEG C of shaking baths with 120r/min training
Foster 24h；
Seed culture based component is：Glucose 25g/L, K2HPO40.2g/L, carbamide 5g/L, Mg2SO41.2g/L, Semen Maydiss
Slurry 35g/L, with 0.1mol/L NaOH pH to 7.0 is adjusted；
S4, by S3 cultivate after thalline with 5% inoculum concentration be inoculated in fermentation medium ferment, fermentation temperature is 35
DEG C, fermentation time 2h obtains glutami acid fermentation liquor；
Fermentation medium components are：Glucose 160g/L, K2HPO4 2.3g/L、Mg2SO41.2g/L, Semen Maydis pulp 12g/L,
Methionine 0.6g/L, VB10.05mg/L；
Mass fraction is added to be 25% carbamide and sodium carbonate, carbamide and sodium carbonate in S5, the glutami acid fermentation liquor in S4
Mass ratio be 1:1, obtain sodium glutamate fermentation liquid；
S6, by the sodium glutamate fermentation liquid for obtaining in S5 with 1700r/min be centrifuged 2min, the feed liquid after centrifugation is passed through
The degerming collection bacteria-removing liquid of micro-filtrate membrane filtration, wherein mocromembrane are 0.2 μm of film, and microfiltration temperature is 30 DEG C, and operation pressure reduction is 0.22MPa；
S7, by the bacteria-removing liquid collected in S6 add decolouring pump in, in bacteria-removing liquid add mass fraction be 1% activated carbon,
Decolouring 20min；
Bacteria-removing liquid containing activated carbon is centrifuged into 10min with 3000r/min, destaining solution is collected；
S8, by the destaining solution obtained in S7 add concentration pan in, carry out concentration in 65 DEG C, obtain sodium glutamate coarse-grain
Body.
Embodiment 2
S1, the corynebacterium glutamicum of slant culture to logarithmic (log) phase is inoculated in broth bouillon in 37 DEG C of shaking baths
Middle culture 16h, shaking speed is 220r/min；
Culture medium is abandoned into supernatant with 3700r/min centrifugation 10min, by the thalline for obtaining pH7.0 phosphate buffer solutions
Thalline is dissolved in pH7.0 phosphate buffer solutions after washing 3 times prepares bacteria suspension, wherein cell concentration is 1 × 108Individual/mL；
S2, obtained bacteria suspension 5mL in S1 is spread evenly across into culture dish bottom carries out microwave treatment, wherein culture dish is straight
Footpath is 9cm, and microwave power is 800W, and pulse frequency is 2450MHz, and microwave treatment time is 60s；
Thalline after microwave treatment is scattered in normal saline and configures bacteria suspension, cell concentration be cell concentration be 1 ×
108Individual/mL；
S3, by the bacterial suspension inoculation obtained in S2 in seed culture medium, in 32 DEG C of shaking baths with 140r/min training
Foster 24h；
Seed culture based component is：Glucose 30g/L, K2HPO40.3g/L, carbamide 6g/L, Mg2SO41.4g/L, Semen Maydiss
Slurry 40g/L, with 0.1mol/L NaOH pH to 7.2 is adjusted；
S4, by S3 cultivate after thalline with 5% inoculum concentration be inoculated in fermentation medium ferment, fermentation temperature is 35
DEG C, fermentation time 3h obtains glutami acid fermentation liquor；
Fermentation medium components are：Glucose 180g/L, K2HPO4 3.5g/L、Mg2SO41.2g/L, Semen Maydis pulp 15g/L,
Methionine 0.9g/L, VB10.1mg/L；
Mass fraction is added to be 25% carbamide and sodium carbonate, carbamide and sodium carbonate in S5, the glutami acid fermentation liquor in S4
Mass ratio be 1:1, obtain sodium glutamate fermentation liquid；
S6, by the sodium glutamate fermentation liquid for obtaining in S5 with 2100r/min be centrifuged 2min, the feed liquid after centrifugation is passed through
The degerming collection bacteria-removing liquid of micro-filtrate membrane filtration, wherein mocromembrane are 0.2 μm of film, and microfiltration temperature is 30 DEG C, and operation pressure reduction is 0.22MPa；
S7, by the bacteria-removing liquid collected in S6 add decolouring pump in, in bacteria-removing liquid add mass fraction be 1% activated carbon,
Decolouring 20min；
Bacteria-removing liquid containing activated carbon is centrifuged into 10min with 3000r/min, destaining solution is collected；
S8, by the destaining solution obtained in S7 add concentration pan in, carry out concentration in 80 DEG C, obtain sodium glutamate coarse-grain
Body.
Embodiment 3
S1, the corynebacterium glutamicum of slant culture to logarithmic (log) phase is inoculated in broth bouillon in 37 DEG C of shaking baths
Middle culture 15h, shaking speed is 220r/min；
Culture medium is abandoned into supernatant with 3500r/min centrifugation 10min, by the thalline for obtaining pH7.0 phosphate buffer solutions
Thalline is dissolved in pH7.0 phosphate buffer solutions after washing 3 times prepares bacteria suspension, wherein cell concentration is 1 × 108Individual/mL；
S2, obtained bacteria suspension 4mL in S1 is spread evenly across into culture dish bottom carries out microwave treatment, wherein culture dish is straight
Footpath is 8cm, and microwave power is 700W, and pulse frequency is 2450MHz, and microwave treatment time is 50s；
Thalline after microwave treatment is scattered in normal saline and configures bacteria suspension, cell concentration be cell concentration be 1 ×
108Individual/mL；
S3, by the bacterial suspension inoculation obtained in S2 in seed culture medium, in 32 DEG C of shaking baths with 130r/min training
Foster 24h；
Seed culture based component is：Glucose 27g/L, K2HPO40.25g/L, carbamide 5.5g/L, Mg2SO4 1.3g/L、
Semen Maydis pulp 37g/L, with 0.1mol/L NaOH pH to 7.1 is adjusted；
S4, by S3 cultivate after thalline with 5% inoculum concentration be inoculated in fermentation medium ferment, fermentation temperature is 35
DEG C, fermentation time 2.5h obtains glutami acid fermentation liquor；
Fermentation medium components are：Glucose 170g/L, K2HPO4 3.0g/L、Mg2SO41.2g/L, Semen Maydis pulp 13g/L,
Methionine 0.8g/L, VB10.75mg/L；
Mass fraction is added to be 25% carbamide and sodium carbonate, carbamide and sodium carbonate in S5, the glutami acid fermentation liquor in S4
Mass ratio be 1:1, obtain sodium glutamate fermentation liquid；
S6, by the sodium glutamate fermentation liquid for obtaining in S5 with 19000r/min be centrifuged 2min, the feed liquid after centrifugation is led to
The degerming collection bacteria-removing liquid of micro-filtrate membrane filtration is crossed, wherein mocromembrane is 0.2 μm of film, and microfiltration temperature is 30 DEG C, and operation pressure reduction is
0.22MPa；
S7, by the bacteria-removing liquid collected in S6 add decolouring pump in, in bacteria-removing liquid add mass fraction be 1% activated carbon,
Decolouring 20min；
Bacteria-removing liquid containing activated carbon is centrifuged into 10min with 3000r/min, destaining solution is collected；
S8, by the destaining solution obtained in S7 add concentration pan in, carry out concentration in 70 DEG C, obtain sodium glutamate coarse-grain
Body.
The invention provides a kind of sodium glutamate production new technique, is educated by carrying out microwave irradiation to corynebacterium glutamicum
Kind, microwave, can be with stimulating protein and nucleotide isopolarity molecule fast vibration used as a kind of electromagnetic wave so that DNA structure is sent out
Changing, so as to hereditary variation occur, substantially increases the acid producing ability of bacterial strain.
Above content is only citing made for the present invention and explanation, and affiliated those skilled in the art are to being retouched
The specific embodiment stated is made various modifications or supplements or substituted using similar mode, without departing from invention or super
More scope defined in the claims, all should belong to protection scope of the present invention.
Claims (9)
1. a kind of sodium glutamate process for separation and purification, it is characterised in that comprise the following specific steps that：
S1, by the corynebacterium glutamicum of slant culture to logarithmic (log) phase be inoculated in broth bouillon cultivate；
Supernatant is abandoned in culture medium centrifugation, the thalline for obtaining is washed 2-3 time with pH7.0 phosphate buffer solutions, thalline is dissolved
Bacteria suspension is prepared in pH7.0 phosphate buffer solutions；
S2, obtained bacteria suspension 3-5mL in S1 is spread evenly across into culture dish bottom carries out microwave treatment, wherein microwave power is
600-800W, pulse frequency is 2450MHz, and microwave treatment time is 45-60s；
Thalline after microwave treatment is scattered in normal saline and configures bacteria suspension；
S3, the bacterial suspension inoculation obtained in S2 is cultivated in seed culture medium；
S4, by S3 cultivate after thalline with 5% inoculum concentration be inoculated in fermentation medium ferment, fermentation temperature be 35 DEG C, send out
Ferment time 2-3h, obtains glutami acid fermentation liquor；
The matter that mass fraction is 25% carbamide and sodium carbonate, carbamide and sodium carbonate is added in S5, the glutami acid fermentation liquor in S4
Amount is than being 1:1, obtain sodium glutamate fermentation liquid；
It is S6, the sodium glutamate fermentation liquid for obtaining in S5 is degerming by micro-filtrate membrane filtration, bacteria-removing liquid is collected, wherein mocromembrane is
0.2 μm of film, microfiltration temperature is 30 DEG C, and operation pressure reduction is 0.22MPa；
S7, by the bacteria-removing liquid collected in S4 add decolouring pump in, in bacteria-removing liquid add mass fraction be 1% activated carbon, decolourize
20min；
By the bacteria-removing liquid collected after centrifugation destaining solution containing activated carbon；
S8, by the destaining solution obtained in S5 add concentration pan in, carry out concentration in 65-80 DEG C, obtain sodium glutamate coarse-grain
Body.
2. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Cultivation temperature in the S1
For 37 DEG C, incubation time is 14-16h, and culture rotating speed is 220r/min.
3. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Centrifugal rotational speed in the S1
For 3200-3700r/min, centrifugation time is 10min.
4. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Described in S1 in bacteria suspension
Cell concentration is 1 × 108Individual/mL, cell concentration is 1 × 10 in bacteria suspension described in S28Individual/mL.
5. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Cultivation temperature is 32 in S3
DEG C, culture rotating speed is 120-140r/min, and incubation time is 24h.
6. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Seed culture described in S3
Based component is：Glucose 25-30g/L, K2HPO40.2-0.3g/L, carbamide 5-6g/L, Mg2SO41.2-1.4g/L, Semen Maydis pulp
35-40g/L, with 0.1mol/L NaOH pH to 7.0-7.2 is adjusted.
7. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Fermentation culture described in S4
Based component is：Glucose 160-180g/L, K2HPO4 2.3-3.5g/L、Mg2SO41.2g/L, Semen Maydis pulp 12-15g/L, first sulfur
Propylhomoserin 0.6-0.9g/L, VB1 0.05-0.1mg/L。
8. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Centrifugation rate described in S6
For 1700-2100r/min, centrifugation time is 2min.
9. a kind of sodium glutamate process for separation and purification according to claim 1, it is characterised in that：Centrifugation rate described in S7
For 3000r/min, centrifugation time is 10min.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109797176A (en) * | 2017-11-17 | 2019-05-24 | 卢松 | A kind of environment-protective process preparing monosodium glutamate |
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| CN101008021A (en) * | 2007-01-23 | 2007-08-01 | 邓毛程 | Glutamic acid fermentation production method supplied with gooey and glucose mixed carbon source matter |
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| CN101491323A (en) * | 2008-11-12 | 2009-07-29 | 山东阜丰生物科技开发有限公司 | New production technique of sodium glutamate |
| CN105039488A (en) * | 2015-09-20 | 2015-11-11 | 呼伦贝尔东北阜丰生物科技有限公司 | Method for extracting gourmet powder through concentration isoelectric point technology |
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