CN107338210B - Abamectin streptomycete synthetic medium and preparation method of fermentation liquor thereof - Google Patents
Abamectin streptomycete synthetic medium and preparation method of fermentation liquor thereof Download PDFInfo
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Abstract
The invention discloses a synthetic medium of streptomyces avermitilis, which is prepared from the following raw materials in parts by weight in a volume of 1L: 8-25 g of maltose monohydrate, 3-14 g of anhydrous glucose, 0.8-2.5 g of threonine, 0.3-0.5 g of magnesium sulfate heptahydrate, 0.005-0.015 g of ferrous sulfate heptahydrate, 0.15-0.45 g of dipotassium phosphate trihydrate, 0.01-0.02 g of manganese sulfate monohydrate, 0.3-0.6 g of sodium chloride, 4-6 g of 3- (N-morpholine) propanesulfonic acid, and the balance of water; also discloses a preparation method of the streptomyces avermitilis fermentation liquor. The synthetic culture medium of the streptomyces avermitilis has reasonable formula matching, rich nutrition and definite culture medium components, is suitable for the growth and production requirements of the streptomyces avermitilis and relevant basic scientific research, and the effective components of the avermectin B1a obtained by the method are 2.84 times of those of the clear culture medium.
Description
Technical Field
The invention belongs to the technical field of microbial culture media and fermentation, and relates to a streptomyces avermitilis synthetic culture medium and a preparation method of fermentation broth thereof.
Background
Streptomyces avermitilis (Streptomyces avermitilis) is a strain for producing avermectins, which was isolated from soil in Jinggang county of Japan in 1978, and the fermentation broth of Streptomyces avermitilis was found to have a high insect-repellent activity by fermentation culture, and then this strain was sent to America Merck for further study. In 2002, the northern research institute of Japan has intensively studied the morphology, physiological characteristics and germ line evolution of the strain, and finally renames it to Streptomyces avermectinus. The Avermectin belongs to gram-positive bacteria of Prokaryotae, Actinomycetes, Streptomycetaceae and Streptomyces.
The avermectin is a macrolide antibiotic generated by fermentation of streptomyces avermitilis, and has wide anthelmintic and insecticidal activities. The fermentation liquor of streptomyces avermitilis contains 8 avermectin components A1a/A1B, B1a/B1B, A2a/A2B and B2a/B2B, wherein the pesticidal effect is optimized with the B1 component, especially the B1a component. The action mechanism of the avermectin is unique, and different from the conventional chemical insecticide, the avermectin drug can cause Cl unrelated to a gamma-aminobutyric acid (GABA) system-The channel is open, the activity of the channel is mainly that the abamectin can generate stereoselective reaction, and the glutamic acid is regulated to control Cl-And the channel allows a large amount of negatively charged chloride ions to flow into the cell, so that the membrane potential is kept in a hyperpolarized state, and the membrane is difficult to depolarize. AVMs are not effective against tapeworms due to the lack of glutamate-controlled chloride channels and the inhibitory neurotransmitter GABA in the tapeworm body. Although the mammal is positioned in the central nervous system by the nerve mediated by gamma-aminobutyric acid, the abamectin is not easy to enter the central nervous system through a blood brain barrier, and the quantity of the medicine distributed to brain tissues of the mammal after entering the body is 100 times lower than that of invertebrate nematodes, so that the abamectin can selectively act on parasites inside and outside a host body and does not act on the host body of the animal, and the abamectin has high selectivity and high safety, is safe to human and livestock at a common dosage, does not harm natural enemies and does not destroy ecology. Due to the outstanding advantages of broad spectrum, high efficiency, no residue, high safety to animals and plants and the like, the abamectin has been widely applied in the fields of agriculture, animal husbandry, medicine and the like, and has wide market prospect and application value.
The abamectin is industrialized in China, and great economic and social benefits are obtained. However, the avermectin production strain has the problems of low fermentation unit, high production cost and the like. How to improve the yield of the abamectin and reduce the production cost is an important subject in abamectin research.
The culture medium used for research and fermentation production of abamectin is a natural culture medium (containing bean cake powder, peanut cake powder and the like), for example: chinese patent application CN10456118A discloses a culture medium for producing doramectin by fermenting mutant streptomyces avermitilis, wherein the culture medium contains soybean cake powder; chinese patent application CN106609288A discloses a method for improving the yield of avermectin B1a in the process of producing avermectin B1a by fermentation of streptomyces avermitilis, wherein the minimal medium of the streptomyces avermitilis comprises bean cake powder. The nutrient-rich culture medium has complex components and rich carbon source and nitrogen source, and has higher titer when used for fermentation. However, due to the complex components of the bean cake powder, the peanut cake powder and the like, the quantitative and qualitative analysis of the experimental results is influenced by the difference of different production places and different batches.
The synthetic culture medium can make up for the defect, and main factors influencing fermentation are determined through research on synthetic culture components, so that the research on a fermentation metabolic mechanism is facilitated, the optimization of the fermentation culture medium is facilitated, and the actual production is promoted. In addition, the synthetic culture medium has definite chemical components and good reproducibility, and can be further applied to various omics and synthetic biology and other comprehensive related basic researches of the streptomyces avermitilis.
Disclosure of Invention
The first technical problem to be solved by the invention is to provide a streptomyces avermitilis synthetic medium. The synthetic medium of the streptomyces avermitilis has reasonable formula matching, rich nutrition and definite medium components, is suitable for the growth and production requirements of the streptomyces avermitilis, and can meet the transcriptomics of the streptomyces avermitilis and other requirements of all-round related researches relating to nutritional growth, metabolism, genetic breeding, production mechanism and the like.
The second technical problem to be solved by the invention is to provide a preparation method of the streptomyces avermitilis fermentation broth. The effective component of the abamectin B1a obtained by the method is 1.22-2.84 times of that of the clear culture medium.
In order to solve the first technical problem, the synthetic culture medium of streptomyces avermitilis of the invention is composed of the following raw materials by weight calculated by 1L volume:
8 to 25g of maltose monohydrate, 3 to 14g of anhydrous glucose, 0.8 to 2.5g of threonine,
0.3 to 0.5g of magnesium sulfate heptahydrate, 0.005 to 0.015g of ferrous sulfate heptahydrate, 0.15 to 0.45g of dipotassium hydrogen phosphate trihydrate,
0.01 to 0.02g of manganese sulfate monohydrate, 0.3 to 0.6g of sodium chloride, 4 to 6g of 3- (N-morpholine) propanesulfonic acid (MOPS for short),
the remainder being water.
Preferably, the pH value of the streptomyces avermitilis synthetic medium is 7.2-7.5.
Preferably, 19.2g of maltose monohydrate, 12.8g of anhydrous glucose, 2g of threonine, 0.5g of magnesium sulfate heptahydrate, 0.01g of ferrous sulfate heptahydrate, 0.3g of dipotassium hydrogen phosphate trihydrate, 0.15g of manganese sulfate monohydrate, 0.5g of sodium chloride and 5g of MOPS.
In order to solve the second technical problem, the invention provides a preparation method of an avermectin streptomyces fermentation liquid, which comprises the following steps:
s1, seed culture
The formula of the seed culture medium is as follows: 30g/L of corn starch, 8g/L of soybean cake powder, 1g/L of peanut cake powder, 4g/L of yeast powder, 0.03g/L of amylase and 0.003g/L of cobalt chloride; adding the rest components into corn starch after gelatinization and hydrolysis, boiling for 15min, cooling at room temperature, and filtering with 10 layers of medical gauze to remove residues, wherein the pH value is 6.86-6.90; the inclined plane is dug and inoculated in a shaking table with 240rpm at the temperature of 28 ℃ for culturing for 35-45 h; centrifuging the mixture in a centrifugal tube at room temperature of 3500-4500 rpm for 5min, discarding the supernatant, washing the mixture with isometric sterile physiological saline, centrifuging the mixture for 3 times, and suspending the mixture with the isometric sterile physiological saline;
s2, preparing a synthetic culture medium of streptomyces avermitilis
The formula of the synthetic culture medium is as follows: 8-25 g/L of maltose monohydrate, 3-14 g/L of anhydrous glucose, 0.8-2.5 g/L of threonine, 0.3-0.5 g/L of magnesium sulfate heptahydrate, 0.005-0.015 g/L of ferrous sulfate heptahydrate, 0.15-0.45 g/L of dipotassium phosphate trihydrate, 0.01-0.02 g/L of manganese sulfate monohydrate, 0.3-0.6 g/L of sodium chloride and 4-6 g/L of 3- (N-morpholine) propanesulfonic acid;
s3 liquid fermentation culture
Transferring the liquid seeds into the synthetic culture medium according to the inoculation amount of 3-10%, and continuously culturing for 7-9 days in a shaking table at 240rpm at 28 ℃ or for 7-9 days in a fermentation tank at the rotating speed of 150rpm and the dissolved oxygen of not less than 30%, thus obtaining the streptomyces avermitilis fermentation liquor containing the abamectin B1 a.
Preferably, in step S1, the seed culture medium formula is: 30g/L of corn starch, 8g/L of soybean cake powder, 10g/L of peanut cake powder, 4g/L of yeast powder, 0.03g/L of amylase and 0.003g/L of cobalt chloride; adding the rest components into corn starch after gelatinization and hydrolysis, boiling for 15min, cooling at room temperature, and filtering with 10 layers of medical gauze to remove residues, wherein the pH value is 6.86-6.90; the slant is dug and inoculated in a shaking table of 240rpm at the temperature of 28 ℃ for 40 h; centrifuging the mixture in a centrifugal tube at the temperature of 28 ℃ and the rpm of 3500-4500 for 5min, discarding the supernatant, washing the mixture with isometric sterile physiological saline, centrifuging the mixture for 3 times, and suspending the mixture with isometric sterile physiological saline;
preferably, in step S3, the liquid seeds are inoculated to the synthetic medium in an amount of 5-8%, and continuously cultured in a shaker at 240rpm at 28 ℃ for 7-9 days or in a fermenter at 150rpm and a dissolved oxygen of not less than 30% for 7-9 days.
Any range recited herein is intended to include the endpoints and any number between the endpoints and any subrange subsumed therein or defined therein.
The starting materials of the present invention are commercially available, unless otherwise specified, and the equipment used in the present invention may be any equipment conventionally used in the art or may be any equipment known in the art.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides a culture medium suitable for producing avermectin by fermentation of streptomyces avermitilis and a preparation method of fermentation liquor of the culture medium. The synthetic culture medium of the streptomyces avermitilis has reasonable formula matching, rich nutrition and definite culture medium components, is suitable for the growth and production requirements of the streptomyces avermitilis, can provide a foundation for various omics, synthetic biology and other comprehensive follow-up researches of the streptomyces avermitilis, and the effective components of the obtained avermectin B1a are 1.22-2.84 times of those of the existing clear culture medium.
Detailed Description
In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.
A synthetic culture medium of streptomyces avermitilis comprises the following components in percentage by weight (g/L): 8-25 parts of maltose monohydrate, 3-14 parts of anhydrous glucose, 0.8-2.5 parts of threonine, 0.3-0.5 part of magnesium sulfate heptahydrate, 0.005-0.015 part of ferrous sulfate heptahydrate, 0.15-0.45 part of dipotassium hydrogen phosphate trihydrate, 0.01-0.02 part of manganese sulfate monohydrate, 0.3-0.6 part of sodium chloride and 4-6 parts of MOPS.
A preparation method of an avermectin streptomyces fermentation liquid comprises the following steps:
s1, seed culture: the formula of the culture medium is (g/L): 30 parts of corn starch, 8 parts of soybean cake powder, 10 parts of peanut cake powder, 4 parts of yeast powder, 0.03 part of amylase and 0.003 part of cobalt chloride; gelatinizing and hydrolyzing starch, adding the rest components, boiling for 15min, cooling at room temperature, and filtering with 10 layers of medical gauze to remove residues; conditions are as follows: shaking-culturing at the initial pH of 6.86-6.90 and 240rpm at 28 ℃ for 40h, centrifuging at 4000rpm for 5min in a 50mL centrifuge tube at room temperature, discarding the supernatant, washing with an equal-volume sterile physiological saline, centrifuging for 3 times, and suspending with the equal-volume sterile physiological saline;
s2, preparing a culture medium for producing the avermectin streptomyces avermitilis strain, wherein the formula of the culture medium is (g/L): 8-25 parts of maltose monohydrate, 3-14 parts of anhydrous glucose, 0.8-2.5 parts of threonine, 0.3-0.5 part of magnesium sulfate heptahydrate, 0.005-0.015 part of ferrous sulfate heptahydrate, 0.15-0.45 part of dipotassium hydrogen phosphate trihydrate, 0.01-0.02 part of manganese sulfate monohydrate, 0.3-0.6 part of sodium chloride and 4-6 parts of MOPS;
s3, liquid fermentation culture: transferring the liquid seeds into the synthetic culture medium according to the inoculation amount of 3-10%, and continuously culturing for 7-9 days at the temperature of 28 ℃ and the rotation speed of a shaking table of 240rpm or under the condition of a 5L fermentation tank with the rotation speed of 150rpm and the dissolved oxygen of not less than 30% for 7-9 days to obtain the streptomycete fermentation liquid containing the abamectin B1a substance.
Example 1
Strain: the Streptomyces avermitilis (Streptomyces avermitilis)9-39 strain is provided by the institute of microorganisms of Chinese academy of sciences.
The composition of the fermentation synthetic medium is (g/L): maltose monohydrate 8, anhydrous glucose 3, threonine 0.8, magnesium sulfate heptahydrate 0.3, ferrous sulfate heptahydrate 0.005, dipotassium hydrogen phosphate trihydrate 0.15, manganese sulfate monohydrate 0.01, sodium chloride 0.3 and MOPS 4.
Firstly, activating a preserved streptomyces avermitilis strain to obtain single strain, inoculating the single strain into a 250mL triangular flask filled with 40mL of filtered seed culture medium, continuously culturing for 40h in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃, obtaining liquid seeds of streptomyces avermitilis, and cleaning and suspending; transferring the liquid seeds into the high-yield liquid culture medium according to the inoculation amount of 5 percent, and continuously culturing for 8 days in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃ to obtain the fermentation liquor of the active substances of the streptomyces avermitilis strain, wherein the yield of the abamectin in the fermentation liquor is 1.22 times that of the existing clear culture medium.
Example 2
Strain: the Streptomyces avermitilis (Streptomyces avermitilis)9-39 strain is provided by the institute of microorganisms of Chinese academy of sciences.
The composition of the fermentation synthetic medium is (g/L): maltose monohydrate 16, anhydrous glucose 8, threonine 1.6, magnesium sulfate heptahydrate 0.3, ferrous sulfate heptahydrate 0.015, dipotassium hydrogen phosphate trihydrate 0.45, manganese sulfate monohydrate 0.02, sodium chloride 0.6 and MOPS 6.
Firstly, activating a preserved streptomyces avermitilis strain to obtain single strain, inoculating the single strain into a 250mL triangular flask filled with 40mL of filtered seed culture medium, continuously culturing for 40h in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃, obtaining liquid seeds of streptomyces avermitilis, and cleaning and suspending; transferring the liquid seeds into the high-yield liquid culture medium according to the inoculation amount of 7 percent, and continuously culturing for 8 days in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃ to obtain the fermentation liquor of the active substances of the streptomyces avermitilis strain, wherein the yield of the abamectin in the fermentation liquor is 2.34 times of that of the existing clear culture medium.
Example 3
Strain: the Streptomyces avermitilis (Streptomyces avermitilis)9-39 strain is provided by the institute of microorganisms of Chinese academy of sciences.
The composition of the fermentation synthetic medium is (g/L): 19.2 parts of maltose monohydrate, 12.8 parts of anhydrous glucose, 2 parts of threonine, 0.5 part of magnesium sulfate heptahydrate, 0.01 part of ferrous sulfate heptahydrate, 0.3 part of dipotassium phosphate trihydrate, 0.15 part of manganese sulfate monohydrate, 0.5 part of sodium chloride and 5 parts of MOPS.
Firstly, activating a preserved streptomyces avermitilis strain to obtain single strain, inoculating the single strain into a 250mL triangular flask filled with 40mL of filtered seed culture medium, continuously culturing for 40h in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃, obtaining liquid seeds of streptomyces avermitilis, and cleaning and suspending; transferring the liquid seeds into the high-yield liquid culture medium according to the inoculation amount of 10%, and continuously culturing for 8 days in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃ to obtain the fermentation liquor of the active substances of the streptomyces avermitilis strain, wherein the yield of the abamectin in the fermentation liquor is 2.84 times that of the existing clear culture medium.
Example 4
Strain: the Streptomyces avermitilis (Streptomyces avermitilis)9-39 strain is provided by the institute of microorganisms of Chinese academy of sciences.
The composition of the fermentation synthetic medium is (g/L): 19.2 parts of maltose monohydrate, 12.8 parts of anhydrous glucose, 2 parts of threonine, 0.5 part of magnesium sulfate heptahydrate, 0.01 part of ferrous sulfate heptahydrate, 0.3 part of dipotassium phosphate trihydrate, 0.15 part of manganese sulfate monohydrate, 0.5 part of sodium chloride and 5 parts of MOPS.
Firstly, activating a preserved streptomyces avermitilis strain to obtain single strain, inoculating the single strain into a 250mL triangular flask filled with 40mL of filtered seed culture medium, continuously culturing for 40h in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃, obtaining liquid seeds of streptomyces avermitilis, and cleaning and suspending; transferring the liquid seeds into the high-yield liquid culture medium according to the inoculation amount of 10%, and continuously culturing for 8 days in a 5L fermentation tank with the rotating speed of 150rpm and the dissolved oxygen of not less than 30% at 28 ℃ to obtain the streptomycete fermentation liquid containing the abamectin B1a substance, wherein the yield of the abamectin in the fermentation liquid is 2.74 times of that of the existing clear culture medium.
Example 5
Strain: the Streptomyces avermitilis (Streptomyces avermitilis)9-39 strain is provided by the institute of microorganisms of Chinese academy of sciences.
The composition of the fermentation synthetic medium is (g/L): 19.2 parts of maltose monohydrate, 12.8 parts of anhydrous glucose, 2 parts of threonine, 0.5 part of magnesium sulfate heptahydrate, 0.01 part of ferrous sulfate heptahydrate, 0.3 part of dipotassium phosphate trihydrate, 0.15 part of manganese sulfate monohydrate, 0.5 part of sodium chloride and 5 parts of MOPS.
Firstly, activating a preserved streptomyces avermitilis strain to obtain single strain, inoculating the single strain into a 250mL triangular flask filled with 40mL of filtered seed culture medium, continuously culturing for 40h in a shaking table at the constant temperature of 240rpm at the temperature of 28 ℃, obtaining liquid seeds of streptomyces avermitilis, and cleaning and suspending; inoculating liquid seeds into the fermentation synthetic medium according to the inoculation amount of 10%, and culturing in a shaker at 28 ℃ and at the constant temperature of 240 rpm; 5mL of fermentation liquor is respectively taken on the 2 nd, 4 th, 6 th and 8 th days of culture and centrifuged at 8000rpm at 4 ℃ for 10min to reserve thalli for transcriptomics analysis, transcription differences of streptomyces avermitilis mRNA in different periods are obtained through IlluminaHiSeq sequencing and result analysis, and a reference basis is provided for analyzing a high-yield mechanism of the streptomyces avermitilis.
It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. Not all embodiments are exhaustive. All obvious changes and modifications which are obvious to the technical scheme of the invention are covered by the protection scope of the invention.
Claims (5)
1. The synthetic culture medium for the streptomyces avermitilis is characterized by comprising the following raw materials in parts by weight in a volume of 1L:
8 to 25g of maltose monohydrate, 3 to 14g of anhydrous glucose, 0.8 to 2.5g of threonine,
0.3 to 0.5g of magnesium sulfate heptahydrate, 0.005 to 0.015g of ferrous sulfate heptahydrate, 0.15 to 0.45g of dipotassium hydrogen phosphate trihydrate,
0.01 to 0.02g of manganese sulfate monohydrate, 0.3 to 0.6g of sodium chloride, 4 to 6g of 3- (N-morpholine) propanesulfonic acid,
the remainder being water.
2. The Streptomyces avermitilis synthetic medium according to claim 1, which is characterized in that: the pH = 7.2-7.5 of the streptomyces avermitilis synthetic medium.
3. The method for preparing the streptomyces avermitilis fermentation broth by using the synthetic medium as claimed in claim 1, which is characterized by comprising the following steps:
s1, seed culture
The formula of the seed culture medium is as follows: 25-35 g/L of corn starch, 6-10 g/L of soybean cake powder, 8-12 g/L of peanut cake powder, 3-5 g/L of yeast powder, 0.02-0.04 g/L of amylase and 0.002-0.004 g/L of cobalt chloride; after gelatinization and hydrolysis of corn starch, adding the rest components, boiling for 12-18 min, cooling at room temperature, and filtering with 8-12 layers of medical gauze to remove residues, wherein the pH is = 6.86-6.90; the inclined plane digging block is inoculated in a shaking table with the temperature of 25-30 ℃ and the rpm of 200-260 for culturing for 35-45 h; centrifuging at 3500-4500 rpm for 4-6 min at 25-30 ℃ in a centrifuge tube, discarding the supernatant, washing with an isometric sterile physiological saline solution, centrifuging for 2-4 times, and suspending with the isometric sterile physiological saline solution;
s2, preparing a synthetic culture medium of streptomyces avermitilis
The formula of the synthetic culture medium is as follows: 8-25 g/L of maltose monohydrate, 3-14 g/L of anhydrous glucose, 0.8-2.5 g/L of threonine, 0.3-0.5 g/L of magnesium sulfate heptahydrate, 0.005-0.015 g/L of ferrous sulfate heptahydrate, 0.15-0.45 g/L of dipotassium phosphate trihydrate, 0.01-0.02 g/L of manganese sulfate monohydrate, 0.3-0.6 g/L of sodium chloride and 4-6 g/L of 3- (N-morpholine) propanesulfonic acid;
s3 liquid fermentation culture
Transferring the liquid seeds into the synthetic culture medium according to the inoculation amount of 3-10%, and continuously culturing for 7-9 days in a shaking table at the temperature of 25-30 ℃ and the rpm of 200-260 or continuously culturing for 7-9 days in a fermentation tank at the rotating speed of 140-160 rpm and the dissolved oxygen content of not less than 30% to obtain the streptomyces avermitilis fermentation liquid containing the abamectin B1a substance.
4. The method for preparing the streptomyces avermitilis fermentation broth according to claim 3, wherein the fermentation broth comprises the following steps:
in step S1, the seed culture medium formula is: 30g/L of corn starch, 8g/L of soybean cake powder, 10g/L of peanut cake powder, 4g/L of yeast powder, 0.03g/L of amylase and 0.003g/L of cobalt chloride; adding the rest components into corn starch after gelatinization and hydrolysis, boiling for 15min, cooling at room temperature, and filtering with 10 layers of medical gauze to remove residues, wherein the pH is = 6.86-6.90; the slant is dug and inoculated in a shaking table of 240rpm at the temperature of 28 ℃ for 40 h; and centrifuging the mixture for 5min at 3500-4500 rpm in a centrifuge tube at 28 ℃, discarding the supernatant, washing the mixture with equal volume of sterile physiological saline, centrifuging the mixture for 3 times, and suspending the mixture with equal volume of sterile physiological saline.
5. The method for preparing the streptomyces avermitilis fermentation broth according to claim 3, wherein the fermentation broth comprises the following steps:
in step S3, liquid seeds are transferred to the synthetic medium according to the inoculum size of 5% -8%, and the liquid seeds are continuously cultured for 7-9 days in a shaking table at 28 ℃ and 240rpm or continuously cultured for 7-9 days in a fermentation tank at the rotation speed of 150rpm and the dissolved oxygen content of not less than 30%.
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| CN105969710A (en) * | 2009-09-25 | 2016-09-28 | Reg生命科学有限责任公司 | Production of fatty acid derivatives |
| CN102634471B (en) * | 2012-04-18 | 2013-09-04 | 南京工业大学 | Abamectin B1a high-yield strain and application thereof |
| US9526267B2 (en) * | 2014-04-17 | 2016-12-27 | Savage River, Inc. | Nutrient-dense meat structured protein products |
| CN105063155B (en) * | 2015-09-25 | 2018-09-11 | 驻马店华中正大有限公司 | Ferment of DM culture medium and the Ferment of DM production method for utilizing the culture medium |
| CN106609288B (en) * | 2015-10-21 | 2020-11-13 | 上海国佳生化工程技术研究中心有限公司 | Method for improving industrial abamectin B by optimizing fermentation medium1aMethod of production of |
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