CN114381386B - Culture medium for producing avermectin through fermentation - Google Patents
Culture medium for producing avermectin through fermentation Download PDFInfo
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- CN114381386B CN114381386B CN202111382342.0A CN202111382342A CN114381386B CN 114381386 B CN114381386 B CN 114381386B CN 202111382342 A CN202111382342 A CN 202111382342A CN 114381386 B CN114381386 B CN 114381386B
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- 238000000855 fermentation Methods 0.000 title claims abstract description 33
- 230000004151 fermentation Effects 0.000 title claims abstract description 33
- 239000005660 Abamectin Substances 0.000 title claims abstract description 15
- RRZXIRBKKLTSOM-XPNPUAGNSA-N avermectin B1a Chemical compound C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 RRZXIRBKKLTSOM-XPNPUAGNSA-N 0.000 title claims abstract description 12
- 239000001963 growth medium Substances 0.000 title abstract description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 52
- 240000008042 Zea mays Species 0.000 claims abstract description 33
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 33
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 33
- 235000005822 corn Nutrition 0.000 claims abstract description 33
- 239000000843 powder Substances 0.000 claims abstract description 30
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 26
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 21
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 16
- 235000019764 Soybean Meal Nutrition 0.000 claims abstract description 10
- 239000004455 soybean meal Substances 0.000 claims abstract description 10
- 229920002261 Corn starch Polymers 0.000 claims abstract description 9
- 229920002472 Starch Polymers 0.000 claims abstract description 9
- 239000008120 corn starch Substances 0.000 claims abstract description 9
- 235000019698 starch Nutrition 0.000 claims abstract description 9
- 239000008107 starch Substances 0.000 claims abstract description 9
- 239000004382 Amylase Substances 0.000 claims abstract description 8
- 102000013142 Amylases Human genes 0.000 claims abstract description 8
- 108010065511 Amylases Proteins 0.000 claims abstract description 8
- 239000003513 alkali Substances 0.000 claims abstract description 8
- 235000019418 amylase Nutrition 0.000 claims abstract description 8
- 229910000019 calcium carbonate Inorganic materials 0.000 claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 8
- 239000013530 defoamer Substances 0.000 claims abstract description 6
- 244000068988 Glycine max Species 0.000 claims description 9
- 235000010469 Glycine max Nutrition 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000011081 inoculation Methods 0.000 claims description 5
- 230000008030 elimination Effects 0.000 claims description 4
- 238000003379 elimination reaction Methods 0.000 claims description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 229910052698 phosphorus Inorganic materials 0.000 claims description 3
- 239000011574 phosphorus Substances 0.000 claims description 3
- 238000011049 filling Methods 0.000 claims description 2
- 230000012010 growth Effects 0.000 abstract description 9
- 241001468227 Streptomyces avermitilis Species 0.000 abstract description 8
- 230000009286 beneficial effect Effects 0.000 abstract description 8
- 241001052560 Thallis Species 0.000 abstract description 5
- IBSREHMXUMOFBB-JFUDTMANSA-N 5u8924t11h Chemical compound O1[C@@H](C)[C@H](O)[C@@H](OC)C[C@@H]1O[C@@H]1[C@@H](OC)C[C@H](O[C@@H]2C(=C/C[C@@H]3C[C@@H](C[C@@]4(O3)C=C[C@H](C)[C@@H](C(C)C)O4)OC(=O)[C@@H]3C=C(C)[C@@H](O)[C@H]4OC\C([C@@]34O)=C/C=C/[C@@H]2C)/C)O[C@H]1C.C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 IBSREHMXUMOFBB-JFUDTMANSA-N 0.000 abstract description 3
- 229950008167 abamectin Drugs 0.000 abstract description 3
- CKUAXEQHGKSLHN-UHFFFAOYSA-N [C].[N] Chemical compound [C].[N] CKUAXEQHGKSLHN-UHFFFAOYSA-N 0.000 abstract description 2
- 230000003698 anagen phase Effects 0.000 abstract description 2
- 230000015572 biosynthetic process Effects 0.000 abstract description 2
- 150000001720 carbohydrates Chemical class 0.000 abstract description 2
- 238000003786 synthesis reaction Methods 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 description 5
- 238000009472 formulation Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 125000001477 organic nitrogen group Chemical group 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000002518 antifoaming agent Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000004904 shortening Methods 0.000 description 2
- 235000017060 Arachis glabrata Nutrition 0.000 description 1
- 244000105624 Arachis hypogaea Species 0.000 description 1
- 235000010777 Arachis hypogaea Nutrition 0.000 description 1
- 235000018262 Arachis monticola Nutrition 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 1
- 230000000895 acaricidal effect Effects 0.000 description 1
- 239000000642 acaricide Substances 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 235000020232 peanut Nutrition 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000001238 wet grinding Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
- C12P19/62—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
- C12P19/623—Avermectin; Milbemycin; Ivermectin; C-076
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- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses a culture medium for producing avermectin by fermentation, which comprises 170-180g/L of corn starch, 15-25g/L of soybean meal, 5-15g/L of yeast powder, 2-10g/L of corn steep liquor, 1-2g/L of light calcium carbonate, 0.1-0.5g/L of solid alkali, 2-6g/L of defoamer and amylase: starch=0.0002-0.0003. The invention increases the dosage of corn steep liquor, reduces the dosage of yeast powder, means that quick-acting nitrogen sources are increased, is more beneficial to the rapid growth and propagation of thalli, shortens the logarithmic growth phase, relatively prolongs the productive phase, and is beneficial to the improvement of fermentation titer. Compared with yeast powder, the corn steep liquor also contains a certain amount of saccharides, the carbon-nitrogen ratio is also improved, more energy support can be provided for continuous synthesis of abamectin by the streptomyces avermitilis, the fermentation titer is more beneficial to increase, the corn steep liquor is turned into wealth, the application of the corn steep liquor is deeply developed, and the added value of the corn steep liquor is further improved.
Description
Technical Field
The invention relates to the technical field of fermentation, in particular to a culture medium for producing abamectin through fermentation.
Background
The avermectin is an insecticidal acaricide obtained by aeration, stirring and submerged fermentation of streptomyces avermitilis in a culture medium containing nutrient elements such as starch, soybean meal, yeast powder, corn steep liquor and the like. As a high-efficiency low-toxicity biological pesticide and a medical intermediate, the novel pesticide is gradually replaced by other pesticides in agriculture. After the industrialized production for nearly 20 years, the market is stable, the process is mature, and in the composition of the manufacturing cost, the raw materials account for about 50% of the cost. In addition to improving the production level, further reducing the cost; the use of inexpensive raw materials instead of expensive raw materials is another way of reducing production costs.
Disclosure of Invention
The invention aims to provide a culture medium for producing avermectin by fermentation aiming at the defects of the prior art.
In order to solve the problems, the invention adopts the following technical scheme:
the culture medium for producing avermectin through fermentation comprises 170-180g/L of corn starch, 15-25g/L of soybean meal, 5-15g/L of yeast powder, 2-10g/L of corn steep liquor, 1-2g/L of light calcium carbonate, 0.1-0.5g/L of solid alkali, 2-6g/L of defoaming agent and amylase: starch=0.0002-0.0003.
Further, the culture medium comprises 174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 5g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025.
Further, the corn steep liquor quality standard: the total nitrogen is more than 10%, the mass concentration of the reducing sugar is more than 2%, the mass fraction of the dissolved phosphorus is less than 1%, the dry matter is more than 45%, and the pH is 3.8-4.2.
Further, the yeast powder quality standard: total nitrogen is more than 6.7 percent, and water content is less than or equal to 8 percent.
Further, the soybean cake has the following quality standard: total nitrogen is more than 6.4% and water content is less than or equal to 8%.
Further, shaking culture conditions: 250mL shaking bottles are adopted, 40mL culture medium is filled, the rotation speed of a shaking table is 250r/min, and the culture temperature is 27-28 ℃.
Further, fermenter culture conditions: volume after elimination is 91-93 m 3 The inoculation amount is 8%, the temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, the stirring frequency is 35-42 Hz.
The beneficial effects of adopting above-mentioned technical scheme to produce lie in:
the main components of the microbial fermentation medium are a carbon source and a nitrogen source, and the nitrogen source refers to a nutrient substance for providing nitrogen for the growth and propagation of microorganisms and synthetic products. Functionally, the method is divided into a quick-acting nitrogen source and a slow-acting nitrogen source, wherein the quick-acting nitrogen source refers to substances with relatively small molecular weight and easy to be utilized by thalli, such as amino acid and ammonium sulfate; the slow-release nitrogen source refers to substances with relatively large molecular weight, such as soybean cake powder, peanut cake powder, cotton seed cake powder and the like. The quick-acting nitrogen source is mainly used for the growth and propagation of thalli in the earlier stage of fermentation, and has great influence on the concentration of the thalli; and the slow-acting nitrogen source mainly maintains the metabolism of the middle-late stage bacteria.
The avermectin is synthesized after the growth and propagation of streptomyces avermitilis reaches a certain stage, the corn steep liquor is a byproduct of preparing corn starch by a wet grinding method, is an inexpensive organic nitrogen source, is rich in amino acid, polypeptide and other nutrient elements, and is beneficial to increasing the dosage of corn steep liquor, reducing the dosage of yeast powder, increasing quick-acting nitrogen sources, being more beneficial to the rapid growth and propagation of thalli, shortening the logarithmic growth phase, relatively prolonging the productive phase and improving the fermentation titer. Compared with yeast powder, the corn steep liquor also contains a certain amount of saccharides, the carbon-nitrogen ratio is also improved, more energy support can be provided for continuous synthesis of abamectin by the streptomyces avermitilis, the fermentation titer is more beneficial to increase, the corn steep liquor is turned into wealth, the application of the corn steep liquor is deeply developed, the added value of the corn steep liquor is further improved, and the production cost is reduced.
Drawings
FIG. 1 is a graph showing the comparison of the titer of example 1 and the average monthly titer in the present invention;
FIG. 2 is a graph showing the comparison of the titer of example 2 with the monthly average titer in the present invention.
Detailed Description
Embodiments of the present invention are described in further detail below with reference to the accompanying drawings and examples. The following examples are illustrative of the invention but are not intended to limit the scope of the invention.
In the description of the present invention, it should be noted that the directions or positional relationships indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
The culture medium for producing avermectin through fermentation comprises 170-180g/L of corn starch, 15-25g/L of soybean meal, 5-15g/L of yeast powder, 2-10g/L of corn steep liquor, 1-2g/L of light calcium carbonate, 0.1-0.5g/L of solid alkali, 2-6g/L of defoaming agent and amylase: starch=0.0002-0.0003.
Further preferably, the culture medium comprises 174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 5g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025.
Further, the corn steep liquor quality standard: the total nitrogen is more than 10%, the mass concentration of the reducing sugar is more than 2%, the mass fraction of the dissolved phosphorus is less than 1%, the dry matter is more than 45%, and the pH is 3.8-4.2; appearance: brown yellow, viscous, aromatic.
Further, the yeast powder quality standard: total nitrogen is more than 6.7 percent, and water content is less than or equal to 8 percent; appearance: pale yellow to yellowish orange powder.
Further, the soybean cake has the following quality standard: total nitrogen is more than 6.4 percent, and water content is less than or equal to 8 percent; appearance: light yellow, no moths and no mildewing.
Further, shaking culture conditions: 250mL shaking bottles are adopted, the filling amount of the culture medium is 40mL, the rotation speed of a shaking table is 250r/min, the culture temperature is 27-28 ℃, 3 shaking bottles are connected to each shaking bottle, and the fermentation titer is an average value of 3 bottles.
Further, fermenter culture conditions: volume after elimination is 91-93 m 3 The inoculation amount is 8%, the temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, the stirring frequency is 35-42 Hz.
The avermectin is a product of secondary fermentation of the streptomyces avermitilis, and is cooled for standby when hypha grows well in a seed tank, namely hypha is gathered into a bar-shaped ball, the bacterial concentration reaches 30% -40%, and the mixed bacteria pollution is avoided.
And calculating the feeding amount of the fermentation tank according to the proportion and the volume after elimination, and preparing materials.
Putting the prepared materials into a tank, metering the volume, cooling to 29-30 ℃ after sterilization, introducing seed liquid with the volume fraction of 8%, ventilating and stirring, sampling and microscopic examination every 8 hours, observing the conditions of sterility and hypha growth, starting to measure the initial titer for 50 hours, and testing every 24 hours until fermentation culture is finished.
The preparation method comprises mixing three nitrogen sources of soybean cake powder, corn steep liquor and yeast powder in a shaker under the condition of unchanged proportion of other raw materials to obtain L9 (3) 4 ) Performing an orthogonal test;
TABLE 1 factor level Table
Remarks: a, soybean cake powder; b corn steep liquor; c yeast powder
Table 2 L9 (3) 4 ) Results of the orthogonal test
The k value data shows that the optimal level combination is A3B2C2, namely 24g/L of soybean meal, 5g/L of corn steep liquor and 8g/L of yeast powder.
The effect of nitrogen source optimization on fermentation titers was verified by the following specific examples. Parallel experiments are carried out on industrialized mass production inconvenient, and average fermentation titers in the month are used as a control.
Example 1
(1) Fermentation cylinder (120 m) 3 ) Formulation of
174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 5g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025, post-consumer volume 91-93 m 3 。
(2) Inoculum size and formulation conditions
The inoculation amount is about 8m 3 The temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, stirring frequency is 35-42 Hz, and culturing time is 357h.
As shown in the data of FIG. 1, FIG. 1 is a graph showing the comparison of the titer and the average monthly titer in example 1, and the average fermentation titer of the soybean cake powder is improved by 7.8% compared with the average monthly level after the nitrogen source is optimized, namely, the soybean cake powder is prepared from 21g/L to 24g/L, the yeast powder is prepared from 10g/L to 8g/L, the corn steep liquor is prepared from 3g/L to 5 g/L. The corn steep liquor is used as a mild quick-acting organic nitrogen source, is more beneficial to the early-stage growth and propagation of the streptomyces avermitilis, is equivalent to shortening the growth and propagation period, prolonging the productive period, further improving the fermentation titer of the streptomyces avermitilis, and repeating the phenomenon after the nitrogen source in the formula is optimized, namely, the tank-placing titer of the streptomyces avermitilis is improved by about 8%.
Example 2
Example 1 shows that increasing the content of quick-acting organic nitrogen source corn steep liquor, reducing the content of soybean cake powder and yeast powder, and improving the pot-release titer by 7.8%; whether the corn steep liquor is added more or not is better, in order to further examine the influence of quick-acting organic nitrogen sources on the tank release titer of the industrialized production of the avermectin, the corn steep liquor is adjusted from 5g/L to 7g/L of the original corn steep liquor, other nitrogen sources and carbon sources are unchanged, and the test result is shown in figure 2
(1) Fermentation cylinder (120 m) 3 ) Formulation of
174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 7g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025, post-consumer volume 91-93 m 3 。
(2) Inoculum size and formulation conditions
The inoculation amount is about 8m 3 The temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, stirring frequency is 35-42 Hz, and culturing time is 357h.
As the data in FIG. 2 show, FIG. 2 is a graph of the titer of example 2 versus the average monthly titer, when the corn steep liquor mass concentration was adjusted from 5g/L to 7g/L, the discharge titer was 5% lower than the control. The method is characterized in that when the quick-acting nitrogen source in the avermectin fermentation formula is too much, mycelium is overgrown and high in fungus concentration are caused, rheological property of fermentation liquor is further influenced, energy and mass transfer in the fermentation liquor are influenced, the growth period is prolonged, the productive period is shortened, and the tank release titer is reduced.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (1)
1. A method for producing avermectin by fermentation is characterized in that the specific fermentation conditions are as follows:
shake cultivation conditions: shaking a bottle by adopting a 250mL bottle, filling 40mL of fermentation medium, rotating a shaking table at the speed of 250r/min, and culturing at the temperature of 27-28 ℃;
fermentation tank culture conditions: 91-93 m percent of volume after elimination, 8 percent of inoculation amount, 27-28 ℃ of temperature and 0.5-0.8 m of air quantity 3 Stirring frequency is 35-42 Hz per minute;
the fermentation medium comprises corn starch 174g/L, soybean meal 24g/L, yeast powder 8g/L, corn steep liquor 5g/L, light calcium carbonate 1.6g/L, solid alkali 0.3g/L, defoamer 4.3g/L and amylase: starch=0.00025;
the corn steep liquor quality standard: the total nitrogen is more than 10%, the mass concentration of the reducing sugar is more than 2%, the mass fraction of the dissolved phosphorus is less than 1%, the dry matter is more than 45%, and the pH is 3.8-4.2;
the yeast powder quality standard: total nitrogen is more than 6.7 percent, and water content is less than or equal to 8 percent;
the quality standard of the soybean cake is as follows: total nitrogen is more than 6.4% and water content is less than or equal to 8%.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101407775A (en) * | 2008-11-27 | 2009-04-15 | 中国科学院微生物研究所 | Method for preparing avermectin and special bacterial strain thereof |
| CN102634471A (en) * | 2012-04-18 | 2012-08-15 | 南京工业大学 | Abamectin B1a high-yield strain and application thereof |
| CN103882080A (en) * | 2014-03-17 | 2014-06-25 | 中国科学院成都生物研究所 | Effective method for preparing avermectin |
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