Fermentation method for increasing aureomycin yield

Abstract

The invention discloses a fermentation method for increasing the yield of aureomycin, belongs to the technical field of biochemical engineering, and can solve the problems that the requirements of the fields of medicines and veterinary drugs on aureomycin are relatively large, and the yield of aureomycin needs to be increased. The method specifically comprises the following steps: inoculating a cultured seed solution into a ferriporphyrin-containing fermentation medium, and fermenting under certain fermentation conditions. By exploring the possibility of supplementing beneficial substances and carrying out fermentation optimization, the yield of the aureomycin is effectively increased, the market demand is met, and a new way is provided for efficient production of the aureomycin.

Description

Technical Field

[0001] The invention relates to the technical field of biochemical engineering, and in particular to a fermentation method for increasing the yield of chlortetracycline. Background Art

[0002] Chlortetracycline, also known as chlortetracycline, plays an important role in the medical field. Chlortetracycline has a broad antibacterial spectrum and has inhibitory effects on Gram-positive bacteria such as Staphylococcus and Pneumococcus, Gram-negative bacteria such as Escherichia coli and Shigella dysenteriae, and is also effective against mycoplasma, Chlamydia, and Rickettsia. The mechanism of action is to bind to the 30S subunit of the bacterial ribosome, hinder the entry of aminoacyl-tRNA into the A position, interfere with peptide chain elongation and protein synthesis, and inhibit bacterial growth. In clinical applications, it is made into eye ointments and eye drops to treat conjunctivitis, keratitis, and trachoma; it is made into ointments that can be used for skin problems such as impetigo, furuncle, and acne; in animal breeding, it can prevent and treat respiratory and intestinal infections in livestock and poultry, and can also promote growth.

[0003] In the field of medicine and veterinary medicine, chlortetracycline plays an irreplaceable role, and it is urgent to increase its production. From a market perspective, with the growth of population and the development of animal husbandry, the demand for chlortetracycline in the treatment of human eye and skin infections, as well as the prevention and control of livestock and poultry diseases continues to rise, and higher production can meet the urgent needs of the market. From a cost perspective, increasing production means reducing unit production costs. Under the production scale effect, equipment depreciation, labor costs, etc. are spread over more products. At the same time, the efficiency of raw material utilization is improved, waste is reduced, and the profit margin of the enterprise can be effectively increased. In the competition of the industry, the increase in production can enable enterprises to respond to the market quickly, win the trust of customers with sufficient supply, and seize market share. Moreover, this also encourages enterprises to continuously optimize production processes and promote technological innovation in the industry, so as to better serve public health and the development of animal husbandry.

[0004] Iron porphyrin is a key component of cytochromes (such as cytochrome c). In the mitochondrial respiratory chain of the cell, between complex III and complex IV of the electron transport chain, the iron porphyrin structure in cytochrome c can accept and transfer electrons. Cytochrome accepts and transfers electrons through the change in the valence state of iron ions in iron porphyrin, thereby realizing the transfer of electrons in the respiratory chain and providing power for the energy metabolism of the cell. The synthesis of biological macromolecules such as proteins, nucleic acids, lipids and polysaccharides in cells requires a lot of energy. Improving energy metabolism can provide sufficient ATP for these synthetic reactions and ensure cell growth, proliferation and repair. The present invention has found through in-depth research that the introduction of iron porphyrin can play its unique role in the fermentation process of chlortetracycline, promote bacterial energy metabolism, improve the utilization of substrates, and thus improve the level of fermentation titer; and because it can reduce the concentration of residual nutrients in the fermentation broth, it can also effectively reduce the difficulty and cost of three wastes treatment. Summary of the invention

[0005] Purpose of the invention: The purpose of the present invention is to provide a fermentation method for increasing the yield of chlortetracycline; it can solve the problem that there is a large demand for chlortetracycline in the medical and veterinary fields, while the yield of chlortetracycline needs to be improved.

[0006] Technical solution: To solve the above technical problems, according to one aspect of the present invention, more specifically, a fermentation method for increasing the yield of chlortetracycline comprises the following steps: inoculating the cultured seed liquid into a fermentation medium, and fermenting under certain fermentation conditions, wherein the fermentation medium contains iron porphyrin.

[0007] Furthermore, the inoculation amount of the seed solution is 1%-20%; the added concentration of the iron porphyrin is 0.1-100 mg / L.

[0008] Furthermore, the inoculation amount of the seed solution is 5%-10%; the added concentration of iron porphyrin is 1-10 mg / L.

[0009] Furthermore, the inoculation amount of the seed solution is 7%; and the added concentration of iron porphyrin is 2-6 mg / L.

[0010] Furthermore, the fermentation medium is composed of: peanut cake powder 1-10, soybean cake powder 1-5, corn starch 1-15, amylase 0.01-0.1, sodium chloride 0.1-1, ammonium sulfate 0.1-5, calcium carbonate 0.1-2, yeast powder 0.1-5, corn steep liquor 0.5-5, magnesium sulfate 0.01-1, potassium dihydrogen phosphate 0.001-0.1, defoamer 0.01-0.1, and the rest is water.

[0011] Furthermore, the fermentation medium is composed of 3-8 peanut cake powder, 2-4 soybean cake powder, 5-10 corn starch, 0.02-0.05 amylase, 0.2-0.7 sodium chloride, 1-3 ammonium sulfate, 0.5-1.5 calcium carbonate, 1-3 yeast powder, 2-4 corn steep liquor, 0.02-0.08 magnesium sulfate, 0.005-0.02 potassium dihydrogen phosphate, 0.02-0.05 defoamer, and the rest is water.

[0012] Furthermore, the fermentation medium is composed of 5 g / 100 mL of peanut cake powder, 3 g of soybean cake powder, 8 g of corn starch, 0.03 g of amylase, 0.4 g of sodium chloride, 2 g of ammonium sulfate, 1 g of calcium carbonate, 2 g of yeast powder, 3 g of corn steep liquor, 0.05 g of magnesium sulfate, 0.01 g of potassium dihydrogen phosphate, 0.04 g of defoaming agent, and the rest is water.

[0013] Furthermore, the fermentation conditions are as follows: pH is controlled at 6.00±0.2, temperature is controlled at 30±1°C, and ventilation ratio is 0.8VVM.

[0014] Beneficial effect: The key technology of the present invention is to inoculate the cultured seed liquid into the fermentation medium containing iron porphyrin and ferment under certain fermentation conditions. Among them, the introduction of iron porphyrin can play a unique role in the fermentation process of chlortetracycline, promote the energy metabolism of the bacteria, improve the utilization of the substrate, and thus improve the level of fermentation titer; and because it can reduce the concentration of residual nutrients in the fermentation liquid, it can also effectively reduce the difficulty and cost of the three wastes treatment. Experiments of the present invention have shown that the yield of chlortetracycline is 55.1% higher than that of conventional technology. DETAILED DESCRIPTION

[0015] In order to make the technical solution of the present invention clearer, the present invention is further described in detail below in conjunction with specific embodiments.

[0016] Example 1

[0017] Spore culture:

[0018] The composition of the slant medium is: 5% bran; 0.01% magnesium sulfate; 0.03% potassium dihydrogen phosphate; 0.04% diammonium hydrogen phosphate; 2% agar powder, and the rest is water. Pick sandy tube spores and inoculate them on the slant medium, culture them at 35±2℃ for about 7 days until the spores mature, and add sterile water to make a spore suspension.

[0019] Seed culture:

[0020] Seed tank culture medium: corn starch 7%; soybean cake powder 5%; calcium carbonate 1%; yeast powder 2%; ammonium sulfate 1%; magnesium sulfate 0.5%; potassium dihydrogen phosphate 0.1%; defoamer 2%, the rest is water. All the above spore suspensions are inoculated into the seed tank, 35±1℃, ventilation ratio 1:1vvm, and cultured for 28 hours.

[0021] Fermentation culture:

[0022] The cultured seed liquid is inoculated into a fermentation medium without iron porphyrin at an inoculation amount of 7%, and fermentation is carried out under certain fermentation conditions. The fermentation medium is composed of 5 peanut cake powder, 3 soybean cake powder, 8 corn starch, 0.03 amylase, 0.4 sodium chloride, 2 ammonium sulfate, 1 calcium carbonate, 2 yeast powder, 3 corn steep liquor, 0.05 magnesium sulfate, 0.01 potassium dihydrogen phosphate, 0.04 defoamer, and the rest is water, and the fermentation conditions are: pH is controlled at 6.00±0.2, temperature is controlled at 30±1°C, ventilation ratio is 0.8VVM, and fermentation cycle is 120 hours.

[0023] The test results at the end of fermentation are as follows:

[0024] name Fermentation titer (u / mL) Residual sugar (g / 100mL) Residual nitrogen (mg / 100mL) Example 1 21085 1.89 648

[0025] Example 2

[0026] The difference from Example 1 is that the fermentation medium contains iron porphyrin, and the added concentration of iron porphyrin is 2 mg / L.

[0027] The test results at the end of fermentation are as follows:

[0028] name Fermentation titer (u / mL) Residual sugar (g / 100mL) Residual nitrogen (mg / 100mL) Example 2 29487 1.76 615

[0029] Example 3

[0030] The difference from Example 1 is that the fermentation medium contains iron porphyrin, and the added concentration of iron porphyrin is 4 mg / L.

[0031] The test results at the end of fermentation are as follows:

[0032] name Fermentation titer (u / mL) Residual sugar (g / 100mL) Residual nitrogen (mg / 100mL) Example 3 32695 1.62 576

[0033] Example 4

[0034] The difference from Example 1 is that the fermentation medium contains iron porphyrin, and the added concentration of iron porphyrin is 6 mg / L.

[0035] The test results at the end of fermentation are as follows:

[0036] name Fermentation titer (u / mL) Residual sugar (g / 100mL) Residual nitrogen (mg / 100mL) Example 4 30258 1.71 598

[0037] According to the above examples, it can be seen that after the addition of iron porphyrin, the residual sugar and ammonia nitrogen in the tank decreased, and the fermentation titer increased. Among them, the titer of Example 3 increased the most, which was 55.1% higher than that of Example 1. The above examples show that the present invention can effectively increase the yield of chlortetracycline, and at the same time effectively reduce the difficulty and cost of the three wastes treatment.

[0038] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A fermentation method for increasing the yield of chlortetracycline, comprising the steps of: inoculating a cultured seed solution into a fermentation medium, and fermenting under certain fermentation conditions, characterized in that: The fermentation medium contains iron porphyrin.

2. A fermentation method for increasing chlortetracycline production according to claim 1, characterized in that: The inoculation amount of the seed solution is 1%-20%; the added concentration of the iron porphyrin is 0.1-100 mg / L.

3. A fermentation method for increasing chlortetracycline production according to claim 2, characterized in that: The inoculation amount of the seed solution is 5%-10%; the added concentration of iron porphyrin is 1-10 mg / L.

4. A fermentation method for increasing chlortetracycline production according to claim 3, characterized in that: The inoculation amount of the seed solution is 7%; the added concentration of iron porphyrin is 2-6 mg / L.

5. A fermentation method for increasing chlortetracycline production according to claim 1, characterized in that: The composition of the fermentation medium is calculated in g / 100 mL: 1-10 peanut cake powder, 1-5 soybean cake powder, 1-15 corn starch, 0.01-0.1 amylase, 0.1-1 sodium chloride, 0.1-5 ammonium sulfate, 0.1-2 calcium carbonate, 0.1-5 yeast powder, 0.5-5 corn steep liquor, 0.01-1 magnesium sulfate, 0.001-0.1 potassium dihydrogen phosphate, 0.01-0.1 defoamer, and the rest is water.

6. A fermentation method for increasing chlortetracycline production according to claim 5, characterized in that: The composition of the fermentation medium is calculated in g / 100 mL: 3-8 peanut cake powder, 2-4 soybean cake powder, 5-10 corn starch, 0.02-0.05 amylase, 0.2-0.7 sodium chloride, 1-3 ammonium sulfate, 0.5-1.5 calcium carbonate, 1-3 yeast powder, 2-4 corn steep liquor, 0.02-0.08 magnesium sulfate, 0.005-0.02 potassium dihydrogen phosphate, 0.02-0.05 defoamer, and the rest is water.

7. A fermentation method for increasing chlortetracycline production according to claim 6, characterized in that: The composition of the fermentation medium is calculated in g / 100mL: 5 peanut cake powder, 3 soybean cake powder, 8 corn starch, 0.03 amylase, 0.4 sodium chloride, 2 ammonium sulfate, 1 calcium carbonate, 2 yeast powder, 3 corn steep liquor, 0.05 magnesium sulfate, 0.01 potassium dihydrogen phosphate, 0.04 defoamer, and the rest is water.

8. A fermentation method for increasing chlortetracycline production according to claim 1, characterized in that: The fermentation conditions are as follows: pH is controlled at 6.00±0.2, temperature is controlled at 30±1°C, and ventilation ratio is 0.8VVM.