CN112592947A - Clean production and fermentation method of neomycin sulfate - Google Patents
Clean production and fermentation method of neomycin sulfate Download PDFInfo
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- CN112592947A CN112592947A CN202011423599.1A CN202011423599A CN112592947A CN 112592947 A CN112592947 A CN 112592947A CN 202011423599 A CN202011423599 A CN 202011423599A CN 112592947 A CN112592947 A CN 112592947A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 90
- 230000004151 fermentation Effects 0.000 title claims abstract description 90
- PGBHMTALBVVCIT-VCIWKGPPSA-N framycetin Chemical compound N[C@@H]1[C@@H](O)[C@H](O)[C@H](CN)O[C@@H]1O[C@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](N)C[C@@H](N)[C@@H]2O)O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CN)O2)N)O[C@@H]1CO PGBHMTALBVVCIT-VCIWKGPPSA-N 0.000 title claims abstract description 28
- 229940053050 neomycin sulfate Drugs 0.000 title claims abstract description 28
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 18
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims abstract description 60
- 239000001963 growth medium Substances 0.000 claims abstract description 40
- 239000002351 wastewater Substances 0.000 claims abstract description 37
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 30
- 235000019270 ammonium chloride Nutrition 0.000 claims abstract description 30
- 239000010802 sludge Substances 0.000 claims abstract description 25
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 15
- 239000007640 basal medium Substances 0.000 claims abstract description 7
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 20
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 20
- 239000011347 resin Substances 0.000 claims description 16
- 229920005989 resin Polymers 0.000 claims description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 239000000725 suspension Substances 0.000 claims description 15
- 229920006395 saturated elastomer Polymers 0.000 claims description 11
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 239000003599 detergent Substances 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 238000001179 sorption measurement Methods 0.000 claims description 10
- 244000068988 Glycine max Species 0.000 claims description 5
- 235000010469 Glycine max Nutrition 0.000 claims description 5
- 239000007836 KH2PO4 Substances 0.000 claims description 5
- 229930193140 Neomycin Natural products 0.000 claims description 5
- 235000007164 Oryza sativa Nutrition 0.000 claims description 5
- 240000008042 Zea mays Species 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 5
- 102000004139 alpha-Amylases Human genes 0.000 claims description 5
- 108090000637 alpha-Amylases Proteins 0.000 claims description 5
- 229940024171 alpha-amylase Drugs 0.000 claims description 5
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 claims description 5
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
- 235000005822 corn Nutrition 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 5
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 235000013312 flour Nutrition 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 229960004927 neomycin Drugs 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 235000009566 rice Nutrition 0.000 claims description 5
- 239000004576 sand Substances 0.000 claims description 5
- 238000011218 seed culture Methods 0.000 claims description 5
- 229920002545 silicone oil Polymers 0.000 claims description 5
- 239000002689 soil Substances 0.000 claims description 5
- 230000001502 supplementing effect Effects 0.000 claims description 5
- 239000008399 tap water Substances 0.000 claims description 5
- 235000020679 tap water Nutrition 0.000 claims description 5
- 239000013587 production medium Substances 0.000 claims 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 abstract description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 abstract description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 abstract description 5
- 235000011130 ammonium sulphate Nutrition 0.000 abstract description 5
- 239000012535 impurity Substances 0.000 abstract description 4
- 231100000331 toxic Toxicity 0.000 abstract description 2
- 230000002588 toxic effect Effects 0.000 abstract description 2
- 238000004065 wastewater treatment Methods 0.000 abstract description 2
- 239000010865 sewage Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 241000209094 Oryza Species 0.000 description 4
- 230000002378 acidificating effect Effects 0.000 description 4
- 235000014113 dietary fatty acids Nutrition 0.000 description 4
- 230000003203 everyday effect Effects 0.000 description 4
- 239000000194 fatty acid Substances 0.000 description 4
- 229930195729 fatty acid Natural products 0.000 description 4
- 150000004665 fatty acids Chemical class 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 208000021959 Abnormal metabolism Diseases 0.000 description 3
- 241000187438 Streptomyces fradiae Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 230000006371 metabolic abnormality Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000020477 pH reduction Effects 0.000 description 3
- 230000004075 alteration Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000002921 fermentation waste Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
Classifications
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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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/46—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin
- C12P19/48—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin the cyclohexyl radical being substituted by two or more nitrogen atoms, e.g. destomycin, neamin
- C12P19/50—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin the cyclohexyl radical being substituted by two or more nitrogen atoms, e.g. destomycin, neamin having two saccharide radicals bound through only oxygen to adjacent ring carbon atoms of the cyclohexyl radical, e.g. ambutyrosin, ribostamycin
- C12P19/52—Preparation of O-glycosides, e.g. glucosides having an oxygen atom of the saccharide radical bound to a cyclohexyl radical, e.g. kasugamycin the cyclohexyl radical being substituted by two or more nitrogen atoms, e.g. destomycin, neamin having two saccharide radicals bound through only oxygen to adjacent ring carbon atoms of the cyclohexyl radical, e.g. ambutyrosin, ribostamycin containing three or more saccharide radicals, e.g. neomycin, lividomycin
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Activated Sludge Processes (AREA)
Abstract
The invention relates to a neomycin sulfate clean production fermentation method, wherein a neomycin sulfate clean production culture medium comprises a fermentation basal culture medium and a supplemented nitrogen source culture medium; wherein the fermentation basal culture medium comprises 0.2-0.73% (w/v) of ammonium chloride, and the supplemented nitrogen source culture medium comprises 28-35% (w/v) of ammonium chloride. The method reduces the amount of sulfate ions in the production wastewater and lightens the toxic effect of the sulfate ions on activated sludge in the subsequent wastewater treatment process under the conditions of not reducing the yield of neomycin sulfate and increasing impurities. The invention carries out fermentation by replacing ammonium sulfate in a fermentation basal medium and a supplementary material by ammonium chloride and controlling the pH value in the fermentation process. The fermented waste water is diluted and pumped into an anaerobic generator filled with sludge. The final fermentation titer and the fermentation components are equivalent to the original ammonium sulfate formula, and the sewage treatment capability of the anaerobic activated sludge is slightly superior. The invention has important significance for the fermentation and clean production of the neomycin sulfate.
Description
Technical Field
The invention relates to the technical field of neomycin sulfate, in particular to a neomycin sulfate clean production fermentation method.
Background
Neomycin sulfate fermentationSulfate ions with higher concentration exist in the wastewater, and the sulfate is reduced by microorganisms in the anaerobic activated sludge to produce S2-It has the functions of inhibiting and poisoning microbe. The main sources of sulfate ions are the medium base material in the fermentation process and ammonium sulfate in the feed.
Disclosure of Invention
In order to solve the technical problems, the invention provides a fermentation method for clean production of neomycin sulfate, ammonium chloride is used for replacing nitrogen source ammonium sulfate, the emission of sulfate ions is reduced from a fermentation source, and S is relieved2-Toxic effects on activated sludge. Meanwhile, the fermentation yield and the impurity ratio of the neomycin sulfate are not increased.
The technical scheme adopted by the invention is that,
a neomycin sulfate clean production culture medium comprises a fermentation basal culture medium and a supplemented nitrogen source culture medium; wherein the fermentation basal culture medium comprises 0.2-0.73% (w/v) of ammonium chloride, and the supplemented nitrogen source culture medium comprises 28-35% (w/v) of ammonium chloride.
Preferably, the fermentation basal medium further comprises: 1-7.0% (w/v) of soybean cake powder, 5.0-9.0% (w/v) of rice flour, 0.1-0.6% (w/v) of corn steep liquor, 0.50-3.0% (w/v) of oral sugar, 0.3-0.6% (w/v) of NaCl0.3-0.6% (w/v) of light calcium carbonate, 0.3-0.6% (w/v) of KH2PO40.01～0.05％(w/v)，Na2HPO40.02-0.10% (w/v), 0.02-0.05% (w/v) of alpha-amylase, and 0.0001-0.3% (w/v) of silicone oil.
The method for performing neomycin sulfate clean production fermentation by using the neomycin sulfate clean production culture medium comprises the following steps:
(1) inoculating sand soil spores to a slant to prepare a slant spore suspension;
(2) inoculating the slant spore suspension into a seed tank filled with a seed culture medium, and performing fermentation culture to obtain a seed solution;
(3) inoculating the seed liquid into a fermentation tank filled with a fermentation basal culture medium, and fermenting by supplementing a supplemented culture medium and ammonium chloride in the fermentation process to obtain a fermentation liquid;
(4) putting the fermentation liquor into an adsorption tank, adding resin to adsorb neomycin, filtering, and collecting wastewater;
(5) collecting the filtered wastewater after adsorption, and treating the filtered wastewater by using activated sludge;
and finishing the clean production of the neomycin sulfate.
Preferably, the bevel spore suspension is obtained by culturing at 26-29 ℃ for 4-8 days in the step (1); the fermentation culture time in the step (2) is 35-40 h.
Preferably, sodium hydroxide solution is used for adjusting the pH value to 6.2-6.4 during the fermentation process in the step (3); the fermentation time is 150-180 h.
Preferably, in the step (3), the ammonia nitrogen concentration of the fermentation liquor is controlled to be 1.4-25g/100ml by feeding ammonium chloride.
Preferably, in the step (4), the saturated resin is washed by using a detergent, low-concentration ammonia water and high-concentration ammonia water in sequence, and the saturated resin is decolored to prepare the product.
More preferably, the detergent contains 0.01-0.25 mol/L hydrochloric acid and 0.01-0.1mol/L ammonium chloride; the saturated resin is washed sequentially with low-concentration ammonia water with the concentration of 0.04-0.4mol/L and high-concentration ammonia water with the concentration of 2.0-3.5 mol/L.
Further preferably, in the step 5), tap water is used for diluting the COD concentration of the wastewater to 4000mg/L, the sludge loading is 0.8-1.2L, the flow rate is 8.5-8.9L/d, the wastewater is treated by activated sludge, the wastewater is replaced once every 2-4 days, the diluted COD is measured before water enters, the treated effluent wastewater is collected every day from the fourth day after sample injection, the COD is measured, and the removal rate of the COD is calculated. Volatile acidic fatty acids (VFAs) of the effluent wastewater were measured.
The invention has the following effects:
1. ammonium chloride is used for replacing nitrogen source ammonium sulfate, so that the emission of sulfate ions is reduced from a fermentation source, and S is relieved2-Toxic effects on activated sludge. Meanwhile, the fermentation yield and the impurity ratio of the neomycin sulfate are not increased.
2. Because the toxic action borne by the activated sludge is reduced, the replacement frequency of the sludge can be further reduced, and the fermentation cost is saved to a certain extent.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention are not limited to the scope of the examples. These examples are intended to illustrate the invention only and are not intended to limit the scope of the invention.
Example 1
A neomycin sulfate clean production culture medium comprises a fermentation basal culture medium and a supplemented nitrogen source culture medium; wherein the nitrogen source feeding medium comprises 28% (w/v) ammonium chloride.
In the fermentation basal medium: soybean cake powder 4.0% (w/v), rice flour 6.0% (w/v), corn steep liquor 0.5% (w/v), oral sugar 1.8% (w/v), ammonium chloride 0.53% (w/v), NaCl0.5% (w/v), light calcium carbonate 0.4% (w/v), KH2PO40.04％(w/v)，Na2HPO40.08% (w/v), 0.04% (w/v) of alpha-amylase and 0.015% (w/v) of silicone oil.
Preferably, the method comprises the steps of:
(1) inoculating sand soil spores to a slant to prepare a slant spore suspension;
(2) inoculating the slant spore suspension into a seed tank filled with a seed culture medium, and performing fermentation culture to obtain a seed solution;
(3) inoculating the seed liquid into a fermentation tank filled with a fermentation basal culture medium, and fermenting by supplementing a supplemented culture medium and ammonium chloride in the fermentation process to obtain a fermentation liquid;
(4) putting the fermentation liquor into an adsorption tank, adding resin to adsorb neomycin, filtering, and collecting wastewater;
(5) collecting the filtered wastewater after adsorption, and treating the filtered wastewater by using activated sludge;
and finishing the clean production of the neomycin sulfate.
Further preferably, the slant spore suspension is obtained by culturing at 26 ℃ for 4 days in the step (1); the time of fermentation culture in the step (2) is 35 h.
Further preferably, the pH is adjusted to 6.2 by using sodium hydroxide solution during the fermentation in the step (3); the fermentation time was 150 h. Resulting in excessive pH reduction of the fermentation liquor and abnormal metabolism of the streptomyces fradiae.
Further preferably, in the step (3), the ammonia nitrogen concentration of the fermentation liquor is controlled to be 1.4g/100ml by feeding ammonium chloride.
Further preferably, in the step (4), the saturated resin is washed with a detergent, low-concentration ammonia water and high-concentration ammonia water in sequence, and the product is decolorized to prepare the product.
Still more preferably, the detergent comprises 0.01mol/L hydrochloric acid and 0.01mol/L ammonium chloride; the saturated resin was washed with a low-concentration ammonia water concentration of 0.04mol/L and a high-concentration ammonia water concentration of 2.0mol/L in this order.
Further preferably, in the step 5), tap water is used for diluting the COD concentration of the wastewater to 4000mg/L, the sludge loading is 0.8L, the flow rate is 8.5L/d, activated sludge is used for treatment, the wastewater is replaced once every 2 days, diluted COD is measured before water enters, treated effluent wastewater is collected every day from the fourth day after sample injection, COD is measured, and the removal rate of the COD is calculated. Volatile acidic fatty acids (VFAs) of the effluent wastewater were measured.
Example 2
A neomycin sulfate clean production culture medium comprises a fermentation basal culture medium and a supplemented nitrogen source culture medium; wherein the nitrogen source feeding medium comprises 35% (w/v) ammonium chloride.
In the fermentation basal medium: soybean cake powder 7.0% (w/v), rice flour 9.0% (w/v), corn steep liquor 0.6% (w/v), oral sugar 3.0% (w/v), ammonium chloride 0.73% (w/v), NaCl0.6% (w/v), light calcium carbonate 0.6% (w/v), KH2PO40.05％(w/v)，Na2HPO40.10% (w/v), alpha-amylase 0.05% (w/v), silicone oil 0.3% (w/v).
Preferably, the method comprises the steps of:
(1) inoculating sand soil spores to a slant to prepare a slant spore suspension;
(2) inoculating the slant spore suspension into a seed tank filled with a seed culture medium, and performing fermentation culture to obtain a seed solution;
(3) inoculating the seed liquid into a fermentation tank filled with a fermentation basal culture medium, and fermenting by supplementing a supplemented culture medium and ammonium chloride in the fermentation process to obtain a fermentation liquid;
(4) putting the fermentation liquor into an adsorption tank, adding resin to adsorb neomycin, filtering, and collecting wastewater;
(5) collecting the filtered wastewater after adsorption, and treating the filtered wastewater by using activated sludge;
and finishing the clean production of the neomycin sulfate.
Further preferably, the bevel spore suspension is obtained by culturing at 26-29 ℃ for 4-8 days in the step (1); the fermentation culture time in the step (2) is 35-40 h.
Further preferably, the pH is adjusted to 6.4 by using sodium hydroxide solution during the fermentation in the step (3); the fermentation time is 180 h. Resulting in excessive pH reduction of the fermentation liquor and abnormal metabolism of the streptomyces fradiae.
Further preferably, in the step (3), the ammonia nitrogen concentration of the fermentation liquor is controlled to be 25g/100ml by feeding ammonium chloride.
Further preferably, in the step (4), the saturated resin is washed with a detergent, low-concentration ammonia water and high-concentration ammonia water in sequence, and the product is decolorized to prepare the product.
Still more preferably, the detergent comprises 0.25mol/L hydrochloric acid and 0.1mol/L ammonium chloride; the saturated resin was washed successively with a low-concentration ammonia water concentration of 0.4mol/L and a high-concentration ammonia water concentration of 3.5 mol/L.
Further preferably, in the step 5), tap water is used for diluting the COD concentration of the wastewater to 4000mg/L, the sludge loading is 1.2L, the flow rate is 8.9L/d, activated sludge is used for treatment, the wastewater is replaced once every 4 days, diluted COD is measured before water enters, treated effluent wastewater is collected every day from the fourth day after sample injection, COD is measured, and the removal rate of the COD is calculated. Volatile acidic fatty acids (VFAs) of the effluent wastewater were measured.
Example 3
A neomycin sulfate clean production culture medium comprises a fermentation basal culture medium and a supplemented nitrogen source culture medium; wherein the nitrogen source feeding medium comprises 32.5% (w/v) ammonium chloride.
In the fermentation basal medium: 7.0% (w/v) of soybean cake powder, 3.6% (w/v) of rice flour, 0.5% (w/v) of corn steep liquor, 2.8% (w/v) of oral sugar, 0.73% (w/v) of ammonium chloride, 0.5% (w/v) of NaCl0.4% (w/v) of light calcium carbonate, KH2PO40.04％(w/v)，Na2HPO40.08% (w/v), 0.04% (w/v) of alpha-amylase and 0.1% (w/v) of silicone oil.
Preferably, the method comprises the steps of:
(1) inoculating sand soil spores to a slant to prepare a slant spore suspension;
(2) inoculating the slant spore suspension into a seed tank filled with a seed culture medium, and performing fermentation culture to obtain a seed solution;
(3) inoculating the seed liquid into a fermentation tank filled with a fermentation basal culture medium, and fermenting by supplementing a supplemented culture medium and ammonium chloride in the fermentation process to obtain a fermentation liquid;
(4) putting the fermentation liquor into an adsorption tank, adding resin to adsorb neomycin, filtering, and collecting wastewater;
(5) collecting the filtered wastewater after adsorption, and treating the filtered wastewater by using activated sludge;
and finishing the clean production of the neomycin sulfate.
Further preferably, the slant spore suspension is obtained by culturing at 28 ℃ for 5 days in the step (1); the time of fermentation culture in the step (2) is 38 h.
Further preferably, the pH is adjusted to 6.3 by using sodium hydroxide solution during the fermentation in the step (3); the fermentation time was 160 h. Resulting in excessive pH reduction of the fermentation liquor and abnormal metabolism of the streptomyces fradiae.
Further preferably, in the step (3), the ammonia nitrogen concentration of the fermentation liquor is controlled to be 15g/100ml by feeding ammonium chloride.
Further preferably, in the step (4), the saturated resin is washed with a detergent, low-concentration ammonia water and high-concentration ammonia water in sequence, and the product is decolorized to prepare the product.
Still more preferably, the detergent comprises 0.15mol/L hydrochloric acid and 0.08mol/L ammonium chloride; the saturated resin was washed successively with a low-concentration ammonia water concentration of 0.3mol/L and a high-concentration ammonia water concentration of 2.5 mol/L.
Further preferably, in the step 5), tap water is used for diluting the COD concentration of the wastewater to 4000mg/L, the sludge loading is 1L, the flow rate is 8.6L/d, activated sludge is used for treatment, the wastewater is replaced once every 3 days, diluted COD is measured before water enters, the treated effluent wastewater is collected every day from the fourth day after sample injection, COD is measured, and the removal rate of the COD is calculated. Volatile acidic fatty acids (VFAs) of the effluent wastewater were measured.
Example 3 was compared with comparative examples, which were cultured in the media shown in tables 1 and 2.
TABLE 1 fermentation basal Medium
TABLE 2 Nitrogen source supplemented Medium
1. Compared with the formula of the comparative example, the ratio of fermentation units to main impurities is basically equivalent, and the quality of the neomycin sulfate is basically consistent before and after change, as shown in the following table 3.
TABLE 3 comparison of fermentation quality of inventive and comparative formulations
2. The results of the activated sludge on wastewater treatment are shown in tables 4 and 5, and the COD removal rate of the formula of the invention is reduced slowly compared with that of the formula of the comparative example, and the sludge treatment capacity is slightly stable.
TABLE 4 fermentation waste water of comparative example formulation
| Number of days | Sample introduction COD mg/L | Effluent CODmg/L | COD removal rate | VFA mM |
| 1 | 12952 | - | - | - |
| 4 | - | 5750 | 55.6％ | 9.95 |
| 5 | - | 3250 | 74.9％ | 7.62 |
| 6 | 11150 | 4050 | 63.6％ | 10.58 |
| 7 | - | 5650 | 49.3％ | 21.58 |
| 8 | - | 4825 | 56.7％ | 19.26 |
| 9 | 10550 | 4797 | 54.5％ | 22.85 |
| 10 | - | 5360 | 49.2％ | 28.14 |
| 11 | 4020 | 61.9％ | 12.48 | |
| 12 | 9940 | 4000 | 59.8％ | 19.68 |
| 13 | 5620 | 43.5％ | 34.70 | |
| 14 | 5506 | 44.6％ | 37.66 | |
| 15 | 11130 | 5392 | 51.6％ | 38.93 |
| 16 | 5652 | 49.2％ | 45.07 | |
| 17 | 6550 | 41.15％ | 48.45 |
TABLE 5 example 3 formulation of fermentation wastewater
The above-described embodiments are merely preferred embodiments of the present invention, and should not be construed as limiting the present invention, and the scope of the present invention is defined by the claims, and equivalents including technical features described in the claims. I.e., equivalent alterations and modifications within the scope hereof, are also intended to be within the scope of the invention.
Claims (9)
1. A neomycin sulfate clean production culture medium is characterized by comprising a fermentation basal culture medium and a supplemented nitrogen source culture medium; wherein the fermentation basal culture medium comprises 0.2-0.73% (w/v) of ammonium chloride, and the supplemented nitrogen source culture medium comprises 28-35% (w/v) of ammonium chloride.
2. The neomycin sulfate clean production medium of claim 1, wherein said fermentation basal medium further comprises: 1-7.0% (w/v) of soybean cake powder, 5.0-9.0% (w/v) of rice flour, 0.1-0.6% (w/v) of corn steep liquor, 0.50-3.0% (w/v) of oral sugar, 0.3-0.6% (w/v) of NaCl0.3-0.6% (w/v) of light calcium carbonate, 0.3-0.6% (w/v) of KH2PO40.01~0.05%（w/v），Na2HPO40.02-0.10% (w/v), 0.02-0.05% (w/v) of alpha-amylase, and 0.0001-0.3% (w/v) of silicone oil.
3. A method for fermentation of neomycin sulfate clean production using the neomycin sulfate clean production medium of any one of claims 1-2, comprising the steps of:
(1) inoculating sand soil spores to a slant to prepare a slant spore suspension;
(2) inoculating the slant spore suspension into a seed tank filled with a seed culture medium, and performing fermentation culture to obtain a seed solution;
(3) inoculating the seed liquid into a fermentation tank filled with a fermentation basal culture medium, and fermenting by supplementing a supplemented culture medium and ammonium chloride in the fermentation process to obtain a fermentation liquid;
(4) putting the fermentation liquor into an adsorption tank, adding resin to adsorb neomycin, filtering, and collecting wastewater;
(5) collecting the filtered wastewater after adsorption, and treating the filtered wastewater by using activated sludge;
and finishing the clean production of the neomycin sulfate.
4. The method according to claim 3, wherein the slant spore suspension is obtained by culturing in step (1) at 26-29 ℃ for 4-8 days; the fermentation culture time in the step (2) is 35-40 h.
5. The method according to claim 3, wherein the pH is adjusted to 6.2-6.4 by using sodium hydroxide solution during the fermentation in the step (3); the fermentation time is 150-180 h.
6. The method according to claim 3, wherein in the step (3), the ammonia nitrogen concentration of the fermentation broth is controlled to be 1.4-25g/100ml by feeding ammonium chloride.
7. The method of claim 3, wherein in the step (4), the saturated resin is washed with a detergent, low-concentration ammonia water and high-concentration ammonia water in sequence, and decolorized to prepare the product.
8. The method of claim 7, wherein the detergent comprises 0.01-0.25 mol/L hydrochloric acid and 0.01-0.1mol/L ammonium chloride; the saturated resin is washed sequentially with low-concentration ammonia water with the concentration of 0.04-0.4mol/L and high-concentration ammonia water with the concentration of 2.0-3.5 mol/L.
9. The method of claim 3, wherein: and 5) diluting the COD concentration of the wastewater to 4000mg/L by using tap water, wherein the sludge loading is 0.8-1.2L, the flow rate is 8.5-8.9L/d, and treating by using activated sludge.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101747409A (en) * | 2010-01-15 | 2010-06-23 | 中鑫东泰(莱阳)纳米基因生物技术有限公司 | Cobra-venom factor and cobra-venom neurotoxin combined separation and purification method |
| CN103074401A (en) * | 2011-12-30 | 2013-05-01 | 宜昌三峡制药有限公司 | Method for producing neomycin sulfate |
| CN104327176A (en) * | 2014-10-24 | 2015-02-04 | 张庆宇 | Extracting method of high-purity cobratoxin and pharmaceutical composition containing high-purity cobratoxin |
| CN107557413A (en) * | 2017-10-16 | 2018-01-09 | 河北圣雪大成制药有限责任公司 | A kind of method for improving the neomycinsulphate quality of production |
| CN110777181A (en) * | 2019-11-26 | 2020-02-11 | 宜昌三峡制药有限公司 | Neomycin sulfate fermentation medium and fermentation method |
| CN110846362A (en) * | 2019-11-26 | 2020-02-28 | 宜昌三峡制药有限公司 | Neomycin sulfate fermentation clean production method |
-
2020
- 2020-12-08 CN CN202011423599.1A patent/CN112592947A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101747409A (en) * | 2010-01-15 | 2010-06-23 | 中鑫东泰(莱阳)纳米基因生物技术有限公司 | Cobra-venom factor and cobra-venom neurotoxin combined separation and purification method |
| CN103074401A (en) * | 2011-12-30 | 2013-05-01 | 宜昌三峡制药有限公司 | Method for producing neomycin sulfate |
| CN104327176A (en) * | 2014-10-24 | 2015-02-04 | 张庆宇 | Extracting method of high-purity cobratoxin and pharmaceutical composition containing high-purity cobratoxin |
| CN107557413A (en) * | 2017-10-16 | 2018-01-09 | 河北圣雪大成制药有限责任公司 | A kind of method for improving the neomycinsulphate quality of production |
| CN110777181A (en) * | 2019-11-26 | 2020-02-11 | 宜昌三峡制药有限公司 | Neomycin sulfate fermentation medium and fermentation method |
| CN110846362A (en) * | 2019-11-26 | 2020-02-28 | 宜昌三峡制药有限公司 | Neomycin sulfate fermentation clean production method |
Non-Patent Citations (3)
| Title |
|---|
| 冯美卿主编: "《生物技术制药》", 31 January 2016 * |
| 刘晓烨主编: "《环境工程微生物学研究技术与方法》", 31 August 2011, 出版发行：哈尔滨：哈尔滨工业大学出版社 * |
| 熊智强等: "利用响应面法优化红谷霉素发酵培养基", 《微生物学通报》 * |
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