CN113265432A - Fermentation medium and application thereof in preparation of amino acid by microbial fermentation - Google Patents
Fermentation medium and application thereof in preparation of amino acid by microbial fermentation Download PDFInfo
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- CN113265432A CN113265432A CN202110697783.3A CN202110697783A CN113265432A CN 113265432 A CN113265432 A CN 113265432A CN 202110697783 A CN202110697783 A CN 202110697783A CN 113265432 A CN113265432 A CN 113265432A
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- 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
- C12P13/18—Glutamic acid; Glutamine using biotin or its derivatives
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention belongs to the technical field of biology, and discloses a fermentation medium which is characterized by comprising a corn steep liquor hydrolysate. According to the invention, the corn steep liquor is hydrolyzed, so that the formula of the fermentation medium is optimized, and the fermentation efficiency is improved.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a fermentation medium and application thereof in preparation of amino acid through microbial fermentation.
Background
In the production process of corn starch, corn kernels need to be soaked in sulfurous acid water, 0.8m3 of soaking water is generated when 1 ton of corn is soaked, the liquid obtained after the soaking water is concentrated is corn steep liquor, and the yield of the corn steep liquor is about 13% of the corn processing amount.
At present, corn steep liquor is mainly used as a culture medium nitrogen source in the fermentation industry. However, with the rapid increase of the processing capacity of the corn in recent years, the supply of the corn steep liquor is larger than the demand, and no other large-dosage use outlet is found. The corn steep liquor contains rich protein, but the Maillard reaction is serious in the processing and concentrating process, so that the corn steep liquor has more complex components, high acidity, high viscosity and dark color. The disposal of corn steep liquor has become the most problematic by-product of corn starch processing enterprises at present.
At present, corn processing enterprises spray corn steep liquor into corn husks (spray corn husks), and the spray corn husks are dark in color, easy to absorb water, absorb moisture and mildew due to serious Maillard reaction. A significant portion of corn processing plants also burn corn steep liquor, using 60 yuan coal for every 1 ton of corn steep liquor burned. Corn starch processing enterprises are also actively looking for new uses for corn steep liquor.
The dry matter of corn steep liquor contains about 45% protein, mainly high-quality albumin and globulin. At present, the deep processing of the corn steep liquor protein on an industrial scale utilizes key technologies to research and develop new technologies, and the demands are urgent. In the production of corn processing 'penta-acid-sugar', corn starch is produced firstly, then amino acid, organic acid and starch sugar are produced by taking starch as raw material, corn steep liquor which is a byproduct is produced in the production process of the corn starch, and the processing amount of the corn steep liquor is about 13% when 1 ton of corn is processed. At present, corn steep liquor is mainly used as a culture medium nitrogen source in the fermentation industry. However, with the rapid expansion of corn processing capacity, the amount of corn steep liquor is not rapidly increased, which results in unreasonable treatment in corn processing enterprises.
At present, corn steep liquor is widely applied to the amino acid fermentation industry, and the technical problem to be solved is how to treat the corn steep liquor so as to improve the application value. Patent CN 107299121A, an amino acid fermentation method using corn steep liquor hydrolysate to replace industrial phosphate, and a method using phytase to hydrolyze corn steep liquor to produce corn steep liquor hydrolysate with free phosphorus content meeting production requirements to replace phosphorus sources such as corn steep liquor and phosphoric acid to be directly used in the fermentation process of amino acid. The invention provides a mode of using corn steep liquor as a raw material and producing corn steep liquor hydrolysate after phytase hydrolysis to replace the corn steep liquor directly used in the common amino acid fermentation production process and adding phosphoric acid as a phosphorus source, and the production index is basically consistent with the normal fermentation.
Disclosure of Invention
The invention aims to provide a fermentation medium and application thereof in preparation of amino acid by microbial fermentation, and aims to improve the utilization efficiency of corn steep liquor, increase the economic benefit of corn processing enterprises and effectively relieve the problem of relative shortage of protein resources.
The invention is realized by the following technical scheme.
A fermentation medium comprising a corn steep liquor hydrolysate.
Further, the air conditioner is provided with a fan,
the fermentation medium comprises: 80g/L glucose, 200g/L corn steep liquor hydrolysate, K2HPO4 2g/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7. mu.g/L.
Further, in the present invention,
the fermentation medium is as follows: 80g/L glucose, 200g/L corn steep liquor hydrolysate, K2HPO4 2g/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7. mu.g/L.
Preferably, the first and second electrodes are formed of a metal,
the preparation method of the fermentation medium comprises the following steps: stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium.
Preferably, the first and second electrodes are formed of a metal,
the corn steep liquor hydrolysate is prepared by the following steps:
adding 1mol/L hydrochloric acid into the corn steep liquor to prepare suspension with the solid content of 20% by mass, and homogenizing by using a high-pressure homogenizer to refine the particle size and control the particle size to be 1-10 microns; heating to 80 deg.C, and performing ultrasonic hydrolysis for 10 min; continuing hydrolyzing for 5h, cooling to 46 ℃, adding ammonia water, adjusting pH to 3.0, adding acidic protease and phytase, performing enzymolysis at 46 ℃ for 10h, inactivating enzyme at 95 ℃ for 3min, filtering with a diatomite filter, discharging residue, adding 1% by weight of active carbon, decolorizing for 45min, filtering to remove active carbon, and freeze-drying at low temperature to obtain dry powder, thus obtaining the corn steep liquor hydrolysate.
More preferably still, the first and second liquid crystal compositions are,
the pressure of the high-pressure homogenizer is 30MPa, the temperature is 32 ℃, and the time is 20 s.
More preferably still, the first and second liquid crystal compositions are,
the ultrasonic frequency is 20 kHz.
More preferably still, the first and second liquid crystal compositions are,
the addition amount of the acid protease is 200u/L, and the addition amount of the phytase is 100 u/L.
The invention also claims the application of the culture medium in the preparation of amino acid by microbial fermentation.
Compared with the prior art, the invention has the advantages of improvement and achievement:
according to the invention, the corn steep liquor is hydrolyzed, so that the formula of the fermentation medium is optimized, and the fermentation efficiency is improved;
the protease is utilized to hydrolyze the corn steep liquor, so that the molecular weight of the protein of the corn steep liquor can be reduced, and the contents of free amino acid and soluble protein are improved;
the corn steep liquor is hydrolyzed by phytase, so that organic phosphorus in the protein of the corn steep liquor can be converted into inorganic phosphorus, the inorganic phosphate in the original culture medium can be completely replaced, the cost is reduced, and the activity of the strain is improved.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present application, the technical solutions in the present application will be clearly and completely described below with reference to specific embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The corn steep liquor hydrolysis process comprises the following steps:
adding 2mol/L hydrochloric acid into the corn steep liquor to prepare suspension with the solid content of 30% by mass, and homogenizing by using a high-pressure homogenizer (the pressure is 30MPa, the temperature is 32 ℃, and the time is 20 s) to refine the particle size and control the particle size to be 1-10 microns; heating to 80 ℃, and hydrolyzing for 10min under the assistance of ultrasonic, wherein the ultrasonic frequency is 20 kHz; continuing hydrolysis for 6h, cooling to 45 ℃, adding ammonia water, adjusting pH to 3.5, adding acid protease (with the addition of 200 u/L) and phytase (100 u/L), hydrolyzing at 45 ℃, carrying out enzymolysis for 9h, inactivating enzyme at 95 ℃ for 3min, filtering with a diatomite filter, discharging residue, adding 1% by weight of active carbon, decolorizing for 30min, filtering to remove the active carbon, and carrying out low-temperature freeze drying to obtain dry powder, thus obtaining the corn steep liquor hydrolysate.
Example 2
The corn steep liquor hydrolysis process comprises the following steps:
adding 1.5mol/L hydrochloric acid into the corn steep liquor to prepare suspension with the solid content of 30% by mass, and homogenizing by using a high-pressure homogenizer (the pressure is 30MPa, the temperature is 30 ℃, and the time is 20 s) to refine the particle size and control the particle size to be 1-10 microns; heating to 85 ℃, and hydrolyzing for 12min under the assistance of ultrasonic, wherein the ultrasonic frequency is 20 kHz; continuing hydrolysis for 7h, cooling to 48 ℃, adding ammonia water, adjusting the pH value to 4.0, adding acid protease (with the addition of 200 u/L) and phytase (100 u/L), carrying out enzymolysis for 12h at the hydrolysis temperature of 48 ℃, inactivating enzyme for 3min at 95 ℃, filtering and deslagging by using a diatomite filter, adding 1% by weight of active carbon for decolorizing for 60min, filtering to remove the active carbon, and carrying out low-temperature freeze drying to obtain dry powder, thus obtaining the corn steep liquor hydrolysate.
Example 3
The corn steep liquor hydrolysis process comprises the following steps:
adding 1mol/L hydrochloric acid into the corn steep liquor to prepare suspension with the solid content of 20% by mass, and homogenizing by using a high-pressure homogenizer (the pressure is 30MPa, the temperature is 32 ℃, and the time is 20 s) to refine the particle size and control the particle size to be 1-10 microns; heating to 80 ℃, and hydrolyzing for 10min under the assistance of ultrasonic, wherein the ultrasonic frequency is 20 kHz; continuing hydrolysis for 5h, cooling to 46 ℃, adding ammonia water, adjusting pH to 3.0, adding acid protease (with the addition of 200 u/L) and phytase (100 u/L), hydrolyzing at 46 ℃, performing enzymolysis for 10h, inactivating enzyme at 95 ℃ for 3min, filtering with a diatomite filter, discharging residue, adding 1% by weight of active carbon, decoloring for 45min, filtering to remove the active carbon, and performing low-temperature freeze drying to obtain dry powder, thus obtaining the corn steep liquor hydrolysate.
Example 4
Fermentation medium 1:
glucose 80g/L, corn steep liquor hydrolyzate (obtained by preparation according to examples 1-3) 200g/L, MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7 mu g/L; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium.
Fermentation Medium 2 (replacement with corn steep liquor, and addition of K is required2HPO4）：
Glucose 80g/L, corn steep liquor, K2HPO4 2g/L，MgSO4·7H2O 50mg/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7 mu g/L; stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium.
The fermentation was carried out using the glutamic acid fermentation strain GDK-9, using the two fermentation media (corn steep liquor hydrolysate in fermentation Medium 1 prepared in example 1), respectively, the rest being the same, and after the fermentation was completed, the glutamic acid yield of fermentation Medium 1 group was 21% higher than that of fermentation Medium 2 group, and no K addition was required2HPO4。
The foregoing list is only illustrative of the preferred embodiments of the present invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (9)
1. A fermentation medium comprising a corn steep liquor hydrolysate.
2. The fermentation medium of claim 1, wherein the fermentation medium comprises: 80g/L glucose, 200g/L corn steep liquor hydrolysate, K2HPO4 2g/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7. mu.g/L.
3. The fermentation medium of claims 1-2, wherein the fermentation medium is: 80g/L glucose, 200g/L corn steep liquor hydrolysate, K2HPO4 2g/L，MnSO4·H2O 3mg/L，FeSO4·7H2O 3mg/L，VB110mg/L, biotin 7. mu.g/L.
4. The fermentation medium of claim 3, wherein the fermentation medium is prepared by: stirring the raw materials uniformly, adjusting pH to 6.5, sterilizing at 121 deg.C for 15min, and naturally cooling to obtain fermentation culture medium.
5. The fermentation medium according to any one of claims 1 to 4, wherein the corn steep liquor hydrolysate is prepared by the following steps:
adding 1mol/L hydrochloric acid into the corn steep liquor to prepare suspension with the solid content of 20% by mass, and homogenizing by using a high-pressure homogenizer to refine the particle size and control the particle size to be 1-10 microns; heating to 80 deg.C, and performing ultrasonic hydrolysis for 10 min; continuing hydrolyzing for 5h, cooling to 46 ℃, adding ammonia water, adjusting pH to 3.0, adding acidic protease and phytase, performing enzymolysis at 46 ℃ for 10h, inactivating enzyme at 95 ℃ for 3min, filtering with a diatomite filter, discharging residue, adding 1% by weight of active carbon, decolorizing for 45min, filtering to remove active carbon, and freeze-drying at low temperature to obtain dry powder, thus obtaining the corn steep liquor hydrolysate.
6. The fermentation medium of claim 5, wherein the high pressure homogenizer has a pressure of 30MPa, a temperature of 32 ℃ and a time of 20 s.
7. The fermentation medium of claim 5, wherein the ultrasonic frequency is 20 kHz.
8. The fermentation medium of claim 5, wherein the acidic protease is added in an amount of 200u/L and the phytase is added in an amount of 100 u/L.
9. Use of a culture medium according to any one of claims 1-8 for the fermentative preparation of an amino acid in a microorganism.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110541014A (en) * | 2019-10-06 | 2019-12-06 | 冯世红 | method for producing tryptophan by using fed-batch culture solution through fermentation |
| CN110592154A (en) * | 2019-10-16 | 2019-12-20 | 冯世红 | Process for producing and extracting tryptophan |
| CN111606756A (en) * | 2020-06-29 | 2020-09-01 | 黄河三角洲京博化工研究院有限公司 | Method for producing liquid fertilizer by using corn steep liquor |
| CN114317349A (en) * | 2021-12-30 | 2022-04-12 | 呼伦贝尔东北阜丰生物科技有限公司 | Preparation method of high-phosphorus composite amino acid nutrient solution |
| CN115323013A (en) * | 2022-09-22 | 2022-11-11 | 内蒙古阜丰生物科技有限公司 | Preparation method of high-quality glutamic acid nitrogen source |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110541014A (en) * | 2019-10-06 | 2019-12-06 | 冯世红 | method for producing tryptophan by using fed-batch culture solution through fermentation |
| CN110592154A (en) * | 2019-10-16 | 2019-12-20 | 冯世红 | Process for producing and extracting tryptophan |
| CN110592154B (en) * | 2019-10-16 | 2023-04-07 | 新疆阜丰生物科技有限公司 | Process for producing and extracting tryptophan |
| CN111606756A (en) * | 2020-06-29 | 2020-09-01 | 黄河三角洲京博化工研究院有限公司 | Method for producing liquid fertilizer by using corn steep liquor |
| CN114317349A (en) * | 2021-12-30 | 2022-04-12 | 呼伦贝尔东北阜丰生物科技有限公司 | Preparation method of high-phosphorus composite amino acid nutrient solution |
| CN115323013A (en) * | 2022-09-22 | 2022-11-11 | 内蒙古阜丰生物科技有限公司 | Preparation method of high-quality glutamic acid nitrogen source |
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