CN114317349A - Preparation method of high-phosphorus composite amino acid nutrient solution - Google Patents
Preparation method of high-phosphorus composite amino acid nutrient solution Download PDFInfo
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- CN114317349A CN114317349A CN202111643845.9A CN202111643845A CN114317349A CN 114317349 A CN114317349 A CN 114317349A CN 202111643845 A CN202111643845 A CN 202111643845A CN 114317349 A CN114317349 A CN 114317349A
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- LPUQAYUQRXPFSQ-DFWYDOINSA-M monosodium L-glutamate Chemical compound [Na+].[O-]C(=O)[C@@H](N)CCC(O)=O LPUQAYUQRXPFSQ-DFWYDOINSA-M 0.000 description 3
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- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 2
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Abstract
The invention belongs to the technical field of biological fermentation, and discloses a preparation method of a high-phosphorus composite amino acid nutrient solution, which comprises the following steps: adding threonine mother liquor into the hydrolyzed corn steep liquor, adding other fermentation medium components, stirring uniformly, and sterilizing with high-pressure steam to obtain the high-phosphorus compound amino acid nutrient solution. The invention can improve the value of the corn steep liquor and reduce the production cost of the amino acid.
Description
Technical Field
The invention belongs to the technical field of biological fermentation, and particularly relates to a preparation method of a high-phosphorus composite amino acid nutrient solution.
Background
Glutamic acid, an acidic amino acid. The molecule contains two carboxyl groups, and the chemical name of the molecule is alpha-aminoglutaric acid. The glutamic acid is colorless crystal and has delicate flavor, is greatly contained in glutamic acid protein, has more content in animal brain, plays an important role in the protein metabolism process in organisms and participates in a plurality of important chemical reactions in animals, plants and microorganisms. Sodium glutamate, commonly known as monosodium glutamate, is the main flavor enhancer and has an enhancing effect on flavor. The sodium glutamate is widely used as a food flavoring agent, can be used independently or used together with other amino acids, and has the function of increasing the flavor when being used in food. At present, the annual output of the glutamic acid is about 200 million tons, various products are exported to more than 50 countries, the output value is about 200 hundred million, the market potential is huge, and the prospect is wide.
A large amount of yeast powder, phosphorus salt, threonine, soybean meal hydrolysate and other raw materials are needed in the glutamic acid fermentation process, and threonine plays an important role in bacterial metabolism. He can protect cell membrane, promote synthesis of phospholipid and oxidation of fatty acid. The metabolism of amino acid in the thallus is balanced, and the bacterium has irreplaceable effects in the growth and acid production stages of the glutamic acid bacteria. The phosphate is an essential element for cell growth, and plays an important role in the synthesis of cell membranes and the supply of energy for cell metabolism. The yeast powder and the soybean meal hydrolysate provide organic nitrogen sources such as amino acid and the like for cell production. Only if the nutrition proportion is balanced, the high-efficiency output and conversion of the production can be ensured. At present, the products are purchased and added into a fermentation culture medium in China, so that the requirements of strain growth and acid production are met. However, these products are expensive, and the production process belongs to the process of high energy consumption and high grain consumption, and the industry of serious environmental pollution.
At present, phosphorus salt, yeast powder, soybean meal hydrolysate, threonine and the like required by glutamic acid fermentation production are purchased outside, the price is high, and the production of the products has large pollution to the environment. Nowadays, glutamic acid production enterprises are increasingly reluctant to compete, high-cost raw material substitution is searched, and the method has strong practical significance in the aspects of reducing the production cost of the enterprises and contributing to environmental protection. Therefore, in the research, effective components of various nitrogen sources are analyzed and quantitatively detected in detail in the existing organic nitrogen source materials, and a composite energy efficient nutrient solution is prepared in various modes of hydrolysis, enzymolysis, acidolysis, compounding and the like in the existing cheap organic nitrogen source, so that part of high-price raw materials can be effectively replaced, and the stability of production indexes is ensured.
With the continuous development of the industry and the continuous application of new technologies, the production indexes in the industry are increasingly improved, and the competition among enterprises is increasingly enthusiastic. How to utilize the existing conditions to research the substitute of high-price raw materials, reduce the consumption of raw materials and improve the competitiveness and the viability of enterprises. The corn steep liquor contains rich protein, amino acid and organic phosphorus, and the current application method only utilizes a small amount of nutrition in the corn steep liquor, most of the nutrition is wasted due to the fact that thalli cannot be directly used, and the sewage treatment cost is high. The corn deep processing industry has great corn steep liquor yield, and if the nutrients which cannot be directly utilized by thalli are decomposed, the nutrients can be utilized, so that the raw material consumption and the production cost of enterprises can be reduced, the treatment cost of environmental protection can also be reduced, and the corn deep processing technology has good environmental protection benefits.
At present, the total amount of the added corn steep liquor in the amino acid fermentation process only accounts for a small part of the corn steep liquor yield, and most of the corn steep liquor is used for preparing fiber feed or compound fertilizer by spraying. Phosphorus content and other molecules provided by the corn steep liquor in amino acid fermentation have great influence on the amino acid fermentation. Therefore, as one of the very cheap and effective organic fermentation components, the method for increasing the use amount of the corn steep liquor in amino acid fermentation and adding exogenous phosphoric acid and a nitrogen source are the most economic ways for increasing the value of corn steep liquor and reducing the production cost of amino acid.
Disclosure of Invention
The invention aims to solve the technical problem of providing a preparation method of a high-phosphorus composite amino acid nutrient solution, aiming at reducing the fermentation cost and improving the utilization value of corn steep liquor.
The invention is realized by the following technical scheme.
A method of processing corn steep liquor, comprising the steps of:
the corn steep liquor is put into a hydrolysis tank for hydrolysis, and the hydrolysis process comprises the following steps: heating to 110-; cooling to 50-60 deg.C, adjusting pH to 2-3, adding acid protease, and performing enzymolysis for 8-12 hr; inactivating enzyme to obtain corn steep liquor.
Preferably, the volume ratio of the sulfuric acid to the corn steep liquor is 1: 1-5.
Preferably, the mass fraction of the sulfuric acid is 50%.
Preferably, the enzyme activity of the acid protease is 5 ten thousand u/g.
Preferably, the amount of the acidic protease added is 0.05% by mass/volume.
On the other hand, the invention also relates to a preparation method of the high-phosphorus composite amino acid nutrient solution, which comprises the following steps: adding 1-3 times volume of threonine mother liquor and other fermentation medium components into the treated corn steep liquor of any one of claims 1-5, stirring, and autoclaving to obtain high-phosphorus compound amino acid nutrient solution.
Preferably, the other fermentation medium components include glucose, inorganic salts and/or vitamins.
More preferably, the vitamin is VB1And/or biotin.
The beneficial effects of the present invention mainly include, but are not limited to, the following:
the method separates phosphorus of phytic acid in the corn steep liquor by combining methods such as a fusion heating method, a sulfuric acid hydrolysis method and a protease hydrolysis enzyme hydrolysis method for absorption and utilization by glutamic acid producing bacteria, thereby achieving the purposes of reducing cost consumption and saving raw materials.
The invention reduces the usage amount of phosphoric acid in the fermentation medium, also reduces the production cost and increases the economic benefit.
The invention increases the dissolved phosphorus in the corn steep from 0.4 per mill to more than 1 per mill; the method is applied to a glutamic acid fermentation culture medium, and can replace the phosphoric acid dosage in the culture medium on the premise of not reducing the fermentation index.
The invention improves the amino nitrogen content of the corn steep liquor by more than 6 percent, has far higher application value than the conventional corn steep liquor product, and can also replace yeast powder.
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
A preparation method of a high-phosphorus composite amino acid nutrient solution comprises the following steps: the corn steep liquor is put into a hydrolysis tank for hydrolysis, and the hydrolysis process comprises the following steps: heating to 120 ℃, preserving heat for 20min, then cooling to 70 ℃, and then adding 50% by mass of sulfuric acid, wherein the volume ratio of the sulfuric acid to the corn steep liquor is 1: 2, stirring and hydrolyzing for 30min at the speed of 100 rpm; reducing the temperature to 50 ℃, adjusting the pH value to 2.5, then adding acid protease (5 ten thousand u/g) with the addition of 0.05 percent of mass-volume ratio, carrying out enzymolysis for 12 hours, and inactivating the enzyme to obtain a corn steep liquor treated substance; adding threonine mother liquor with the same volume into the treated corn steep liquor, adding other components, stirring uniformly, and sterilizing with high-pressure steam to obtain a glutamic acid fermentation culture medium, namely the high-phosphorus composite amino acid nutrient solution.
Comparative example 1
A preparation method of a high-phosphorus composite amino acid nutrient solution comprises the following steps: the corn steep liquor is put into a hydrolysis tank for hydrolysis, and the hydrolysis process comprises the following steps: adding 50% of sulfuric acid by mass, wherein the volume ratio of the sulfuric acid to the corn steep liquor is 1: 2, stirring and hydrolyzing for 30min at the speed of 100 rpm; reducing the temperature to 50 ℃, adjusting the pH value to 2.5, then adding acid protease (5 ten thousand u/g) with the addition of 0.05 percent of mass-volume ratio, carrying out enzymolysis for 12 hours, and inactivating the enzyme to obtain a corn steep liquor treated substance; adding threonine mother liquor with the same volume into the treated corn steep liquor, adding other components, stirring uniformly, and sterilizing with high-pressure steam to obtain a glutamic acid fermentation culture medium, namely the high-phosphorus composite amino acid nutrient solution.
Comparative example 2
A preparation method of a high-phosphorus composite amino acid nutrient solution comprises the following steps: the corn steep liquor is put into a hydrolysis tank for hydrolysis, and the hydrolysis process comprises the following steps: heating to 120 ℃, keeping the temperature for 20min, then cooling to 50 ℃, adjusting the pH to 2.5, then adding acid protease (5 ten thousand u/g), wherein the addition amount is 0.05 percent by mass volume ratio, the enzymolysis time is 12h, and inactivating the enzyme to obtain a corn steep liquor treatment product; adding threonine mother liquor and other components into the treated corn steep liquor, stirring uniformly, and sterilizing with high-pressure steam to obtain glutamic acid fermentation culture medium, namely the high-phosphorus compound amino acid nutrient solution.
Comparative example 3
A preparation method of a high-phosphorus composite amino acid nutrient solution comprises the following steps: the corn steep liquor is put into a hydrolysis tank for hydrolysis, and the hydrolysis process comprises the following steps: adding 50% of sulfuric acid by mass, wherein the volume ratio of the sulfuric acid to the corn steep liquor is 1: 2, stirring and hydrolyzing for 30min at the speed of 100 rpm; cooling to 50 deg.C, adjusting pH to 2.5, adding acidic protease (5 ten thousand u/g) in an amount of 0.05% by mass/volume, performing enzymolysis for 12h, heating to 120 deg.C, and keeping the temperature for 20min to obtain corn steep liquor; adding threonine mother liquor with the same volume into the treated corn steep liquor, adding other components, stirring uniformly, and sterilizing with high-pressure steam to obtain a glutamic acid fermentation culture medium, namely the high-phosphorus composite amino acid nutrient solution.
Example 2
The corn steep liquor used in the invention is the same batch of product, and the dry matter of the corn steep liquor accounts for 40%. Threonine mother liquor (wet organic matter 3.29%, COD 4.06g/L, total nitrogen 367 mg/L) obtained in a Fufeng threonine fermentation production workshop is used.
The components of different glutamic acid fermentation culture mediums are as follows:
group 1: 80g/L glucose, 45ml/L corn steep liquor treated in example 1, 45ml/L threonine mother liquor, MnSO4·H2O 3mg/L，FeSO4·7H2O 3 mg/L，VB110mg/L, biotin 7 mu g/L and pH 6.9-7.1;
group 2: glucose 80g/L, comparative example 1 Jade45ml/L of rice pulp treatment product, 45ml/L of threonine mother liquor and MnSO4·H2O 3mg/L，FeSO4·7H2O 3 mg/L，VB110mg/L, biotin 7 mu g/L and pH 6.9-7.1;
group 3: 80g/L glucose, 30ml/L corn steep liquor treated in comparative example 2, 45ml/L threonine mother liquor, MnSO4·H2O 3mg/L，FeSO4·7H2O 3 mg/L，VB110mg/L, biotin 7 mu g/L and pH 6.9-7.1;
group 4: glucose 80g/L, corn steep liquor treated product 45ml/L of comparative example 3, threonine mother liquor 45ml/L, MnSO4·H2O 3mg/L，FeSO4·7H2O 3 mg/L，VB110mg/L, biotin 7 mu g/L and pH 6.9-7.1;
group 5: 80g/L glucose, 45ml/L corn steep liquor, 45ml/L threonine mother liquor and MnSO4·H2O 3mg/L，FeSO4·7H2O 3 mg/L，VB110mg/L, biotin 7 mu g/L and pH 6.9-7.1;
the process for producing the glutamic acid by fermenting the culture medium comprises the following steps:
inoculating Corynebacterium glutamicum (CGMCC No.5481 as example) with 5% inoculum size into 500L full-automatic fermentation tank containing 300L fermentation medium for fermentation culture, wherein the inoculation concentration of thallus is OD600nmFermenting for 48h when the fermentation time is 0.8, and collecting fermentation liquor;
in the whole fermentation process, the fermentation temperature is controlled to be 38 ℃, the ventilation ratio is 1: 0.6, the stirring speed is 400r/min, the dissolved oxygen is maintained at 20 percent, glucose with the fed-batch mass percentage concentration of 80 percent is maintained at 1.5 percent of residual sugar, and the fed-batch defoaming agent is used for defoaming;
example 3
The results of the comparison of the fermentation of different media and the content of dissolved phosphorus in corn steep liquor in example 2 are shown in Table 1.
TABLE 1
| Corn steep liquor type | Dissolved phosphorus g/L | Glutamic acid g/L |
| Group 1 | 1.13 | 153 |
| Group 2 | 0.67 | 139 |
| Group 3 | 1.18 | 142 |
| Group 4 | 0.83 | 145 |
| Group 5 | 0.41 | 121 |
The invention aims at solving the problem that the corn steeping concentrated solution decomposes organic phosphorus into inorganic phosphorus in a single or combined mode of heating, adding sulfuric acid or adding enzyme and the like, and finds the optimal process route by adjusting different process parameters and the treatment sequence of hydrolysis methods (heating, adding acid, adding alkali and adding enzyme).
The corn dipping concentrated solution contains about 1% -3% of phytic acid, and because the phosphoric acid is combined with inositol, the phosphorus cannot be utilized by thalli, so that the waste of the phosphorus is caused, and the environment is also polluted. Through different hydrolysis modes, organic phosphorus is decomposed into inorganic phosphorus to be utilized by thalli, the addition of phosphoric acid in a fermentation culture medium is reduced, and the cost is reduced.
Meanwhile, the corn steep liquor contains rich protein, the bacteria can not utilize macromolecular protein in the fermentation process, only micromolecular amino acid and functional peptide can be utilized, the macromolecular protein is hydrolyzed into micromolecular amino acid and functional peptide in a hydrolysis mode, and the fermentation effect is obviously better than that of the conventional corn steep liquor product.
As shown in Table 1 above, groups 1 and 3 had the highest dissolved phosphorus content, but group 3 had no acid dilution, less the effect of the acid dilution, and group 3 had a dissolved phosphorus concentration of about 0.8g/L, much lower than group 1, at the same volume. Likewise, group 1 resulted in increased yields of fermentation products due to increased levels of dissolved phosphorus and small amino acids and functional peptides.
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 (8)
1. A method of processing corn steep liquor, comprising the steps of:
the corn steep liquor is put into a hydrolysis tank for hydrolysis, and the hydrolysis process comprises the following steps: heating to 110-; cooling to 50-60 deg.C, adjusting pH to 2-3, adding acid protease, and performing enzymolysis for 8-12 hr; inactivating enzyme to obtain corn steep liquor.
2. The method of claim 1, wherein the volume ratio of sulfuric acid to corn steep liquor is 1: 1-5.
3. The method according to claim 1, wherein the mass fraction of sulfuric acid is 50%.
4. The method according to claim 1, wherein the acidic protease has an enzymatic activity of 5 wu/g.
5. The method according to claim 1, wherein the acidic protease is added in an amount of 0.05% by mass/volume.
6. A preparation method of a high-phosphorus composite amino acid nutrient solution comprises the following steps: adding 1-3 times volume of threonine mother liquor and other fermentation medium components into the treated corn steep liquor of any one of claims 1-5, stirring, and autoclaving to obtain high-phosphorus compound amino acid nutrient solution.
7. The method of claim 6, wherein the other fermentation medium components comprise glucose, inorganic salts, and/or vitamins.
8. The method according to claim 7, wherein the vitamin is VB1And/or biotin.
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