CN108774637A - A method of producing nimoctin - Google Patents
A method of producing nimoctin Download PDFInfo
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- CN108774637A CN108774637A CN201810101138.9A CN201810101138A CN108774637A CN 108774637 A CN108774637 A CN 108774637A CN 201810101138 A CN201810101138 A CN 201810101138A CN 108774637 A CN108774637 A CN 108774637A
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- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/16—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing two or more hetero rings
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Abstract
The present invention provides a kind of methods of high efficiency production nimoctin, fermenting and producing is carried out under Valine existence condition using the cyaneogriseus streptomyces of anti-Valine, solve the problems, such as that fermentation middle and later periods fermentation yield increasess slowly in traditional handicraft, fermentation period is extended, the fermentation production rate of nimoctin is significantly improved.
Description
Technical field
The present invention relates to a kind of microbial fermentation processes more particularly to a kind of sides that nimoctin fermentation yield can be improved
Method.
Background technology
Nimoctin (Nemadectin) is a kind of ten hexa-atomic macrolide antibiotics, belongs to the one of mibemycin race
Member, by cyaneogriseus streptomyces (Streptomyces cyaneogriseus sp.Noncyanogenus) fermented generation, Buddhist nun's Mack
Spit of fland can obtain the stronger antibiotic Moses gram of activity by the semi-synthetic positions C-23 introducing=N-OCH groups in its molecular structure
Spit of fland (Moxidectin).The structural formula of nimoctin is：
It has been used with anthelmintic initially as animal since moxidectin mid-term the 1980s, can efficiently kill line
Worm and epizoon etc., while having good safety to animal, be a kind of novel wide spectrum, efficiently, in the big ring of safety
Esters expelling parasite antibiotic, has been widely used for veterinary clinic at present, and the market demand increasingly increases.Therefore, it is possible to high efficiency production
The method of nimoctin is desired.
Invention content
In order to improve the production efficiency of nimoctin, the present invention provides a kind of methods of high efficiency production nimoctin.
The method of production nimoctin provided by the present invention, including：Contain cyaneogriseus streptomyces (Streptomyces
Cyaneogriseus sp.Noncyanogenus) zymotic fluid in, be added Valine, ferment.Wherein, described bluish grey
Streptomycete is the cyaneogriseus streptomyces of anti-Valine；During the fermentation, cyaneogriseus streptomyces growth factor matter in zymotic fluid is controlled
Amount concentration maintains 0.1%-8%.
In an advantageous embodiment, cyaneogriseus streptomyces (the Streptomyces cyaneogriseus
Sp.Noncyanogenus) KS12-08, the bacterial strain are deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms
The heart, preservation date on 2 15th, 2017, deposit number CGMCC No.13661.
In an advantageous embodiment, it is preferably 0.01%- that the Valine being added, which accounts for zymotic fluid weight ratio,
0.1%, more preferably 0.02%-0.08%, such as：0.04%, 0.05%, 0.06%, 0.07%.
In an advantageous embodiment, in the cyaneogriseus streptomyces fermentation process, Valine is added daily, is added daily
Valine to account for zymotic fluid weight ratio be preferably 0.01%-0.1%, more preferably 0.02%-0.08%, such as：0.04%,
0.05%, 0.06%, 0.07%.
In an advantageous embodiment, Valine is added after completing primary metabolite in zymotic fluid in cyaneogriseus streptomyces.
In an advantageous embodiment, it at least in the fermentation process after Valine is added, controls blue in zymotic fluid
Grey streptomycete growth factor mass concentration maintains 0.2%-6%, more preferably 0.4%-4%, more preferably 0.5%-2%,
More preferably 0.7%-1.5%, such as 1%, 1.2%.
In an advantageous embodiment, in the fermentation process, when cyaneogriseus streptomyces growth factor mass concentration is less than
When 1.5%, carry out adding cyaneogriseus streptomyces growth factor.
Wherein, the cyaneogriseus streptomyces growth factor is preferably one or more of monosaccharide and disaccharide or oligosaccharides.Wherein,
The monosaccharide can be one or more of preferably Su Li sugar, xylose, glucose, fructose, mannose, galactolipin；It is described
Disaccharide can be preferably one or more of sucrose, maltose, isomaltose, lactose, trehalose；The oligosaccharides is preferably
Any one or a few in oligofructose, xylo-oligosaccharide, oligomeric maltose, oligoisomaltose.
In an advantageous embodiment, in the fermentation process, fermentation temperature is preferably 20-35 DEG C, more preferably 25-33
DEG C, such as 28 DEG C, 30 DEG C.
In an advantageous embodiment, the cyaneogriseus streptomyces seed liquor is seeded in fermentation medium, is cultivated, is obtained
Obtain the zymotic fluid.
In an advantageous embodiment, on the basis of the fermentation medium total weight, the fermentation medium includes：Portugal
Grape sugar 8%, maltodextrin 4%, soybean cake powder 2.8%, yeast powder 0.5%, magnesium sulfate 0.1%, zinc sulfate 0.01%, calcium carbonate
0.4%.
It is highly preferred that when glucose quality concentration is less than 1.5% in zymotic fluid, carry out adding glucose.
In an advantageous embodiment, the cyaneogriseus streptomyces spore inoculating obtains seed liquor in seed culture medium.
In an advantageous embodiment, on the basis of the seed culture medium total weight, the seed culture medium includes：Wheat
Bud dextrin 2.0%, glucose 1.0%, yeast powder 1.0%, soybean cake powder 1.5%, magnesium sulfate 0.1%, dipotassium hydrogen phosphate
0.1%, calcium carbonate 0.2%.
In an advantageous embodiment, spore suspension is made in the cyaneogriseus streptomyces, in the culture containing Valine
It is cultivated in base, further carries out bacterial strain and culture that resistance screening obtains tolerance Valine, obtain spore.
The method of production nimoctin provided by the invention solves the middle and later periods fermentation yield that ferments in traditional handicraft and increases
Slow problem, extends fermentation period, significantly improves the fermentation production rate of nimoctin.
Specific implementation mode
Initial strain is conventional cyaneogriseus streptomyces, by initial strain by inclined-plane culture, after with slant strains spore is made
Suspension, it is 10 to be diluted to spore concentration8Dithyl sulfate solution is added into suspension by a/ml, then puts 28 DEG C of shakes in shaking table
Culture is swung, taking out 5ml spore suspensions with liquid-transfering gun is placed in the culture dish of 6cm diameters, is put into stirrer.Then by ultraviolet
Fluorescent tube irradiates mutagenesis.
Spore suspension after ultraviolet mutagenesis is diluted to certain concentration gradient under red light, takes 0.2ml to be coated with respectively
In on culture medium flat plate, 28 DEG C are protected from light culture 7-8 days, and picking single bacterium colony screens to obtain the bacterial strain of tolerance Valine, entitled
Cyaneogriseus streptomyces (Streptomyces cyaneogriseus sp.Noncyanogenus) KS12-08, during which is deposited in
State's Microbiological Culture Collection administration committee common micro-organisms center, preservation date on 2 15th, 2017, deposit number CGMCC
No.13661。
16S rDNA Sequence Detections are the results show that mutagenic strain CGMCC No.13661 belong to cyaneogriseus streptomyces.This is lured
Change bacterial strain CGMCC No.13661 fibrillaes of spores are short, enclose spiral shape in fine and close 2-3, spore is in oval to short cylindrical, surface light
It is sliding；Gram formula synthesizes in No. 1 agar medium：Gas silk is powdery, and base silk becomes later dusty blue from initially colourless；It is resistant to L- figured silk fabrics
Propylhomoserin.
Embodiment 1
Step 1, cyaneogriseus streptomyces KS12-08 is inoculated in slant medium, in 28~30 DEG C of temperature, humidity 20~60%
Under the conditions of cultivate 7~8 days, obtain ripe slant strains；Slant medium includes：Starch 1.0%, glucose 0.5%, yeast
Powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%, agar 2.0%；
Step 2, appropriate spore inoculating is scraped in liquid seed culture medium from ripe slant strains, in temperature 28~30
DEG C, it is cultivated 26~30 hours under the conditions of 200~500rpm of speed of agitator, obtains ripe seed liquor；Seed culture medium includes：Wheat
Bud dextrin 2.0%, glucose 1.0%, yeast powder 1.0%, soybean cake powder 1.5%, magnesium sulfate 0.1%, dipotassium hydrogen phosphate
0.1%, calcium carbonate 0.2%；
Step 3, ripe seed liquor is seeded to according to 8~12% inoculum concentration in fermentation medium, temperature 28~
30 DEG C, 200~600rpm of speed of agitator, tank presses 0.02~0.06MPa, fermented and cultured under conditions of dissolved oxygen >=30% on tank；Hair
Ferment culture medium includes：Glucose 8%, maltodextrin 4%, soybean cake powder 2.8%, yeast powder 0.5%, magnesium sulfate 0.1%, sulfuric acid
Zinc 0.01%, calcium carbonate 0.4%；
Valine is configured to the solution of debita spissitudo, after sterilizing, in fermentation process, fills into account in zymotic fluid daily
The Valine of zymotic fluid 0.05wt%.
Embodiment 2
Step 1, cyaneogriseus streptomyces KS12-08 is inoculated in slant medium, in 28~30 DEG C of temperature, humidity 20~60%
Under the conditions of cultivate 7~8 days, obtain ripe slant strains；Slant medium includes：Starch 1.0%, glucose 0.5%, yeast
Powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%, agar 2.0%；
Step 2, appropriate spore inoculating is scraped in liquid seed culture medium from ripe slant strains, in temperature 28~30
DEG C, it is cultivated 26~30 hours under the conditions of 200~500rpm of speed of agitator, obtains ripe seed liquor；Seed culture medium includes：Wheat
Bud dextrin 2.0%, glucose 1.0%, yeast powder 1.0%, soybean cake powder 1.5%, magnesium sulfate 0.1%, dipotassium hydrogen phosphate
0.1%, calcium carbonate 0.2%；
Step 3, ripe seed liquor is seeded to according to 8~12% inoculum concentration in fermentation medium, temperature 28~
30 DEG C, 200~600rpm of speed of agitator, tank presses 0.02~0.06MPa, fermented and cultured under conditions of dissolved oxygen >=30% on tank；Hair
Ferment culture medium includes：Glucose 8%, maltodextrin 4%, soybean cake powder 2.8%, yeast powder 0.5%, magnesium sulfate 0.1%, sulfuric acid
Zinc 0.01%, calcium carbonate 0.4%；
When fermenting to 48h, the Valine of 0.05wt% is filled into zymotic fluid daily, Valine is made into concentration
The solution of 5wt% after sterilizing, daily fills into Valine solution in fermentation tank according to the 1wt% of fermentating liquid volume.
Embodiment 3
Step 1, cyaneogriseus streptomyces KS12-08 is inoculated in slant medium, in 28~30 DEG C of temperature, humidity 20~60%
Under the conditions of cultivate 7~8 days, obtain ripe slant strains；Slant medium includes：Starch 1.0%, glucose 0.5%, yeast
Powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%, agar 2.0%；
Step 2, appropriate spore inoculating is scraped in liquid seed culture medium from ripe slant strains, in temperature 28~30
DEG C, it is cultivated 26~30 hours under the conditions of 200~500rpm of speed of agitator, obtains ripe seed liquor；Seed culture medium includes：Wheat
Bud dextrin 2.0%, glucose 1.0%, yeast powder 1.0%, soybean cake powder 1.5%, magnesium sulfate 0.1%, dipotassium hydrogen phosphate
0.1%, calcium carbonate 0.2%；
Step 3, ripe seed liquor is seeded to according to 8~12% inoculum concentration in fermentation medium, temperature 28~
30 DEG C, 200~600rpm of speed of agitator, tank presses 0.02~0.06MPa, fermented and cultured under conditions of dissolved oxygen >=30% on tank；Hair
Ferment culture medium includes：Glucose 8%, maltodextrin 4%, soybean cake powder 2.8%, yeast powder 0.5%, magnesium sulfate 0.1%, sulfuric acid
Zinc 0.01%, calcium carbonate 0.4%；
When fermenting to 48h, the Valine of 0.05wt% is filled into zymotic fluid daily, Valine is made into concentration
The solution of 5wt% after sterilizing, daily fills into Valine solution in fermentation tank according to the 1wt% of fermentating liquid volume.
Concentration of glucose in tank top fermentation liquid is measured by sampling, when concentration of glucose is reduced to 1.5% or less, starts to mend Portugal
Grape sugar juice maintains the concentration of glucose in tank top fermentation liquid between 0.5~2.0%, and 12h stops mending sugar before putting tank.
Terminate fermentation after 240~260h of fermentation, obtains the zymotic fluid containing nimoctin.
Embodiment 4
Step 1, cyaneogriseus streptomyces KS12-08 is inoculated in slant medium, in 28~30 DEG C of temperature, humidity 20~60%
Under the conditions of cultivate 7~8 days, obtain ripe slant strains；Slant medium includes：Starch 1.0%, glucose 0.5%, yeast
Powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%, agar 2.0%；
Step 2, the strain inclined plane grown is taken, the spore inoculating on the inclined-planes scraping about 0.5cm × 0.5cm to 500ml kinds
In sub- shaking flask, seed flask loading amount is 50ml, the seed flask being inoculated with 28 DEG C of cultures on the shaking table that rotating speed is 200rpm
28h。
Step 3, shake-flask seed will have been cultivated to be seeded on 5L glass fermentation tanks according to 8% inoculum concentration, the dress of fermentation tank
Amount is 2.5L, and fermentation tank culture condition is 28 DEG C, ventilatory capacity 0.8vvm, speed of agitator 200rpm of tank temperature, is passed through in fermentation process
Speed of agitator is improved, oxygen dissolving value (DO) >=30% on fermentation tank is controlled.
When fermented and cultured is to 48h, start the Valine solution for mending the 5wt% concentration to have sterilized, mends daily once, every time
25ml is mended, i.e., respectively fills into a Valine when fermentation is to 48h, 72h, 96h, 120h, 144h, 168h, 192h, 216h.
When fermented and cultured is to 150h, starts stream plus mend glucose solution, Portugal in sugared speed control zymotic fluid is mended by adjusting
Grape sugar concentration stops mending sugar between 0.5~2.0%, when 228h.
Terminate to cultivate when fermented and cultured is to 240h, obtains the zymotic fluid containing nimoctin.
Embodiment 5
Step 1, cyaneogriseus streptomyces KS12-08 is inoculated in slant medium, in 28~30 DEG C of temperature, humidity 20~60%
Under the conditions of cultivate 7~8 days, obtain ripe slant strains；Slant medium includes：Starch 1.0%, glucose 0.5%, yeast
Powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%, agar 2.0%；
Step 2, the strain inclined plane grown is taken, the spore inoculating on the inclined-planes scraping about 0.5cm × 0.5cm to 500ml kinds
In sub- shaking flask, seed flask loading amount is 50ml, the seed flask being inoculated with 28 DEG C of cultures on the shaking table that rotating speed is 200rpm
28h。
Step 3, shake-flask seed will have been cultivated to be seeded on 10L stainless steel seeding tanks according to 1% inoculum concentration, seeding tank
Loading amount is 5.0L, and seed tank culture condition is 28 DEG C, ventilatory capacity 1.0vvm, speed of agitator 200rpm of tank temperature, is led in incubation
Raising speed of agitator is crossed, oxygen dissolving value (DO) >=30% on seeding tank is controlled, the seed tank culture time is for 24 hours.
Step 4, seeding tank seed liquor will have been cultivated to be seeded on 50L fermentation tanks according to 10% inoculum concentration, fermentation tank
Loading amount is 30L, and fermentation tank culture condition is 28 DEG C, ventilatory capacity 0.8vvm, speed of agitator 200rpm of tank temperature, is passed through in incubation
Speed of agitator is improved, oxygen dissolving value (DO) >=30% on fermentation tank is controlled.
When fermented and cultured is to 48h, start the Valine solution for mending the 5wt% concentration to have sterilized, at the uniform velocity flow feeding
Mode, the amount of filling into is 300ml/ days, and when 228h stops feed supplement.
When fermented and cultured is to 150h, starts stream plus mend glucose solution, Portugal in sugared speed control zymotic fluid is mended by adjusting
Grape sugar concentration stops mending sugar between 0.5~2.0%, when 228h.
Terminate to cultivate when fermented and cultured is to 240h, obtains the zymotic fluid containing nimoctin.
Comparative example 1
Step 1, initial cyaneogriseus streptomyces (bacterial strain that mutagenesis is penetrated without UV illumination) are inoculated in slant medium,
28~30 DEG C of temperature is cultivated 7~8 days under the conditions of humidity 20~60%, and ripe slant strains are obtained；Slant medium includes：
Starch 1.0%, glucose 0.5%, yeast powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%,
Agar 2.0%；
Step 2, the strain inclined plane grown is taken, the spore inoculating on the inclined-planes scraping about 0.5cm × 0.5cm to 500ml kinds
In sub- shaking flask, seed flask loading amount is 50ml, the seed flask being inoculated with 28 DEG C of cultures on the shaking table that rotating speed is 200rpm
28h。
Step 3, shake-flask seed will have been cultivated to be seeded on 10L stainless steel seeding tanks according to 1% inoculum concentration, seeding tank
Loading amount is 5.0L, and seed tank culture condition is 28 DEG C, ventilatory capacity 1.0vvm, speed of agitator 200rpm of tank temperature, is led in incubation
Raising speed of agitator is crossed, oxygen dissolving value (DO) >=30% on seeding tank is controlled, the seed tank culture time is for 24 hours.
Step 4, seeding tank seed liquor will have been cultivated to be seeded on 50L fermentation tanks according to 10% inoculum concentration, fermentation tank
Loading amount is 30L, and fermentation tank culture condition is 28 DEG C, ventilatory capacity 0.8vvm, speed of agitator 200rpm of tank temperature, is passed through in incubation
Speed of agitator is improved, oxygen dissolving value (DO) >=30% on fermentation tank is controlled.
When fermented and cultured is to 48h, start the Valine solution for mending the 5wt% concentration to have sterilized, at the uniform velocity flow feeding
Mode, the amount of filling into is 300ml/ days, and when 228h stops feed supplement.
Terminate to cultivate when fermented and cultured is to 240h, obtains the zymotic fluid containing nimoctin.
Comparative example 2
Step 1, initial cyaneogriseus streptomyces (bacterial strain that mutagenesis is penetrated without UV illumination) are inoculated in slant medium,
28~30 DEG C of temperature is cultivated 7~8 days under the conditions of humidity 20~60%, and ripe slant strains are obtained；Slant medium includes：
Starch 1.0%, glucose 0.5%, yeast powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%,
Agar 2.0%；
Step 2, the strain inclined plane grown is taken, the spore inoculating on the inclined-planes scraping about 0.5cm × 0.5cm to 500ml kinds
In sub- shaking flask, seed flask loading amount is 50ml, the seed flask being inoculated with 28 DEG C of cultures on the shaking table that rotating speed is 200rpm
28h。
Step 3, shake-flask seed will have been cultivated to be seeded on 10L stainless steel seeding tanks according to 1% inoculum concentration, seeding tank
Loading amount is 5.0L, and seed tank culture condition is 28 DEG C, ventilatory capacity 1.0vvm, speed of agitator 200rpm of tank temperature, is led in incubation
Raising speed of agitator is crossed, oxygen dissolving value (DO) >=30% on seeding tank is controlled, the seed tank culture time is for 24 hours.
Step 4, seeding tank seed liquor will have been cultivated to be seeded on 50L fermentation tanks according to 10% inoculum concentration, fermentation tank
Loading amount is 30L, and fermentation tank culture condition is 28 DEG C, ventilatory capacity 0.8vvm, speed of agitator 200rpm of tank temperature, is passed through in incubation
Speed of agitator is improved, oxygen dissolving value (DO) >=30% on fermentation tank is controlled.
When fermented and cultured is to 48h, start the Valine solution for mending the 5wt% concentration to have sterilized, at the uniform velocity flow feeding
Mode, the amount of filling into is 300ml/ days, and when 228h stops feed supplement.
When fermented and cultured is to 150h, starts stream plus mend glucose solution, Portugal in sugared speed control zymotic fluid is mended by adjusting
Grape sugar concentration stops mending sugar between 0.5~2.0%, when 228h.
Terminate to cultivate when fermented and cultured is to 240h, obtains the zymotic fluid containing nimoctin.
Comparative example 3
Step 1, initial cyaneogriseus streptomyces (bacterial strain that mutagenesis is penetrated without UV illumination) are inoculated in slant medium,
28~30 DEG C of temperature is cultivated 7~8 days under the conditions of humidity 20~60%, and ripe slant strains are obtained；Slant medium includes：
Starch 1.0%, glucose 0.5%, yeast powder 0.4%, magnesium sulfate 0.05%, dipotassium hydrogen phosphate 0.05%, calcium carbonate 0.1%,
Agar 2.0%；
Step 2, the strain inclined plane grown is taken, the spore inoculating on the inclined-planes scraping about 0.5cm × 0.5cm to 500ml kinds
In sub- shaking flask, seed flask loading amount is 50ml, the seed flask being inoculated with 28 DEG C of cultures on the shaking table that rotating speed is 200rpm
28h。
Step 3, shake-flask seed will have been cultivated to be seeded on 10L stainless steel seeding tanks according to 1% inoculum concentration, seeding tank
Loading amount is 5.0L, and seed tank culture condition is 28 DEG C, ventilatory capacity 1.0vvm, speed of agitator 200rpm of tank temperature, is led in incubation
Raising speed of agitator is crossed, oxygen dissolving value (DO) >=30% on seeding tank is controlled, the seed tank culture time is for 24 hours.
Step 4, seeding tank seed liquor will have been cultivated to be seeded on 50L fermentation tanks according to 10% inoculum concentration, fermentation tank
Loading amount is 30L, and fermentation tank culture condition is 28 DEG C, ventilatory capacity 0.8vvm, speed of agitator 200rpm of tank temperature, is passed through in incubation
Speed of agitator is improved, oxygen dissolving value (DO) >=30% on fermentation tank is controlled.
Terminate to cultivate when fermented and cultured is to 240h, obtains the zymotic fluid containing nimoctin.
Table 1, the present invention produce nimoctin method and comparative example Contrast on effect
Bacterial strain used herein is can be seen that by the comparison of table 1, exists in Valine, can significantly carry
High nimoctin fermentation yield, non-mutagenic strain (comparative example 1-3) instead result in nimoctin hair after Valine is added
Ferment yield declines.This is because conventional cyaneogriseus streptomyces have certain sensibility to Valine, it can be to indigo plant in the presence of Valine
Grey streptomycete generates toxicity, and strain activity is caused to decline, to which fermentation efficiency declines
It is applicant's understanding that the isopropyl in nimoctin molecule comes from Valine, therefore added in zymotic fluid
Valine can save time and nutrition needed for thalline itself synthesis Valine, to improve yield, for this purpose, the application
The cyaneogriseus streptomyces filtered out can resist Valine toxicity, after Valine is added, can carry production nimoctin
Yield.For example, 4-5 of the embodiment of the present invention is compared with comparative example 1-3, and the method that the present invention generates nimoctin, resulting yield ratio
Non- mutagenic strain maximum output improves 9-12%.
Comparative example 1 and embodiment 2,3 in initial fermentation medium it is found that add the effect of Valine not
Obviously, and after the 48h that ferments, bacterial strain primary metabolite is basically completed, and cometabolism occupies main status, adds Valine at this time
Then there are preferable effect, the effect especially added on a small quantity by several times more excellent.
Comparative example 1-5, the present invention maintain the concentration of the growth factors such as glucose during the fermentation, can also be into one
Step improves fermentation efficiency.
It should be understood that various culture mediums described in the above embodiment of the present invention, can also be carried out with other culture mediums
It substitutes, technical difficulty is not present in this to those skilled in the art.
Specific embodiments of the present invention are described in detail above, but it is intended only as example, the present invention is simultaneously unlimited
It is formed on particular embodiments described above.To those skilled in the art, it is any to the equivalent modifications that carry out of the present invention and
It substitutes also all among scope of the invention.Therefore, without departing from the spirit and scope of the invention made by impartial conversion and
Modification, all should be contained within the scope of the invention.
Claims (10)
1. a kind of method producing nimoctin, which is characterized in that including：In zymotic fluid containing cyaneogriseus streptomyces, L- is added
Valine ferments；The cyaneogriseus streptomyces are the cyaneogriseus streptomyces of anti-Valine；During the fermentation, control fermentation
Cyaneogriseus streptomyces growth factor mass concentration maintains 0.1%-8% in liquid.
2. the method for production nimoctin according to claim 1, which is characterized in that during the cyaneogriseus streptomyces are deposited in
State's Microbiological Culture Collection administration committee common micro-organisms center, preservation date on 2 15th, 2017, deposit number CGMCC
No.13661。
3. the method for production nimoctin according to claim 1, which is characterized in that the Valine being added accounts for fermentation
Liquid weight ratio is 0.01%-0.1%.
4. the method for production nimoctin according to claim 3, which is characterized in that the cyaneogriseus streptomyces fermentation process
In, Valine is added daily, and it is 0.01%-0.1% that the Valine added daily, which accounts for zymotic fluid weight ratio,.
5. the method for production nimoctin according to claim 1, which is characterized in that cyaneogriseus streptomyces are complete in zymotic fluid
At Valine is added after primary metabolite.
6. the method for production nimoctin according to claim 1, which is characterized in that the fermentation after Valine is added
In the process, it controls cyaneogriseus streptomyces growth factor mass concentration in zymotic fluid and maintains 0.5%-2%.
7. the method for production nimoctin according to claim 6, which is characterized in that in the fermentation process, when bluish grey
When streptomycete growth factor mass concentration is less than 1.5%, carry out adding cyaneogriseus streptomyces growth factor.
8. the method for the production nimoctin described according to claim 6 or 7, which is characterized in that the cyaneogriseus streptomyces growth
The factor is one or more of monosaccharide and disaccharide or oligosaccharides；Wherein, the monosaccharide be Su Li sugar, xylose, glucose, fructose,
One or more of mannose, galactolipin；The disaccharide is one in sucrose, maltose, isomaltose, lactose, trehalose
Kind is several；The oligosaccharides be oligofructose, xylo-oligosaccharide, oligomeric maltose, in oligoisomaltose any one or it is several
Kind.
9. the method for production nimoctin according to claim 1, which is characterized in that the cyaneogriseus streptomyces KS12-08
Seed liquor is seeded in fermentation medium, is cultivated, and the zymotic fluid is obtained；Using the fermentation medium total weight as base
Standard, the fermentation medium include：Glucose 8%, maltodextrin 4%, soybean cake powder 2.8%, yeast powder 0.5%, magnesium sulfate
0.1%, zinc sulfate 0.01%, calcium carbonate 0.4%.
10. the method for production nimoctin according to claim 8, which is characterized in that in the fermentation process, fermentation temperature
Degree is 20-35 DEG C.
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| CN102336796A (en) * | 2010-07-27 | 2012-02-01 | 北大方正集团有限公司 | Preparation method of nemadectin |
| CN104745654A (en) * | 2013-12-27 | 2015-07-01 | 牡丹江佰佳信生物科技有限公司 | Method and strain for preparing Nemadectin |
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| CN102336796A (en) * | 2010-07-27 | 2012-02-01 | 北大方正集团有限公司 | Preparation method of nemadectin |
| CN104745654A (en) * | 2013-12-27 | 2015-07-01 | 牡丹江佰佳信生物科技有限公司 | Method and strain for preparing Nemadectin |
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