CN1683521A - Agricultural antibiotic strain for producing fungus killer and its preparing process - Google Patents
Agricultural antibiotic strain for producing fungus killer and its preparing process Download PDFInfo
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- CN1683521A CN1683521A CN 200510046029 CN200510046029A CN1683521A CN 1683521 A CN1683521 A CN 1683521A CN 200510046029 CN200510046029 CN 200510046029 CN 200510046029 A CN200510046029 A CN 200510046029A CN 1683521 A CN1683521 A CN 1683521A
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Abstract
The present invention belongs to the field of farm chemical producing technology. The strain is kabicidin streptomyce with preservation number of CGMCC No. 1247, and generates tetraene antibiotic of molecular expression of C35H53NO13 and molecular weight of 695. The compound is used in preventing and controlling fungus diseases of crops. The fermentation process includes the following successive steps: 1. strain propagation via inoculating the slant seed to seed tank for culture at 26-32 deg.c pH 6.0-8.0 and pressure 0.05-0.2 MPa for 40-50 hr, and sterilizing for 30 min; 2. creating fermenting antibiotics; and 3. post-fermentation treatment. The present invention may be used in controlling several kinds one fungus diseases while ensuring human and animal safety and protecting environment.
Description
One, technical field:
The invention belongs to pesticide production technology field, particularly relate to and a kind ofly produce the bacterial strain of mycocidal agricultural antibiotic and produce agricultural antibiotic with it, and the preparation technology of this agricultural antibiotic.
Two, background technology:
Agricultural chemicals is to guarantee the farm crop stable and high yields, solves the indispensable valuable cargo of world food problem, also is an industry that can obtain tremendous economic interests and social benefit simultaneously.Yet problems such as the residual caused environmental pollution of chemical pesticide, person poultry poisoning, pathogenic bacteria or pest resistance to insecticide enhancing are on the rise, and study energetically, develop, production and applying biological agricultural chemicals, have become the new trend of global agricultural chemicals industry development.Countries in the world are taking practical steps and are reducing the use of chemical pesticide at present, once announce to cancel the registration of 91 kinds of chemical pesticides as EPA; European Union has formulated 5 years and the ten year plan that reduces half chemical pesticide usage quantity, and forbids using chemical pesticide in the agricultural-food productions such as gourd, fruit and vegetable of immediate access.China also will develop biological pesticide and green food is listed China's Agenda 21 in.Therefore, efficient, the low toxicity of research and development,, disease and pest good with the environment compatibility novel agricultural microbiotic that is difficult for producing resistance will have vast market prospect more.
Agricultural antibiotic has advantages such as more efficient, quick, safe as the important monoid in the biological pesticide than other biological pesticide.Since Japan in 1958 successfully developed miewensu and is used to prevent and treat rice blast, states such as Japan and the United States, English, method had carried out the research and development work of agricultural antibiotic in succession, and obtain very big achievement.Frequent day by day along with science and technology development and International Academic Exchange, the R﹠D work of China aspect agricultural antibiotic obtains very big achievement.Also develop some agricultural antibiotics that have independent intellectual property right, efficient, wide spectrum such as jingganmycin, middle living rhzomorph, Gongzhuling mycin, pesticide corrosion 120 etc., and all in crop pest control work, bringing into play important effect.But the share of market of China's biological pesticide only has 2%～3% at present, and its potentiality are far from obtaining sufficient development and application.
Existing agricultural antibiotic is mainly used in the prevention and control of plant diseases, pest control of land for growing field crops farm crop such as paddy rice, wheat, the biological pesticide that is used to prevent and treat particularly fungal disease on the fruit and vegetables such as cucumber downy mildew, blight, leaf muld of tomato, gray mold, capsicum epidemic disease, grapevine anthracnose etc. is then less relatively, therefore, at the important disease on the fruit and vegetables, the novel agricultural microbiotic of research and development broad-spectrum high efficacy, in China and even crucial meaning is all arranged in the world, this invention is formulated under this background.
Three, summary of the invention:
1, goal of the invention: the invention provides a kind of bacterial strain and zymotechnique thereof that produces antifungal agricultural antibiotics, its purpose is effectively controlling plant fungal disease, can ensure person poultry safety and protection environment simultaneously.
2, technical scheme: the present invention is achieved through the following technical solutions:
A kind of bacterial strain that produces agricultural antibiotic, its formal name used at school are kabicidin streptomycete Streptomycesfungicidicus Yh06, and its culture presevation is numbered CGMCC NO.1247; This strain cell wall belongs to amino acid I type, and its full cell carbohydrate type belongs to sugared type C.
A kind of agricultural antibiotic is characterized in that: its chemical structural formula is:
Its molecular formula is C
35H
53NO
13, its molecular weight is 695.
The preparation technology of above-mentioned agricultural antibiotic is characterized in that: this technology is carried out as follows successively:
(A) bacterial classification expands numerous process: adopt kabicidin streptomycete Streptomyces fungicidicusYh06, add following basis and cultivate composition, percentage composition meter by weight in seeding tank:
Groundnut meal 2.0～4.0;
Soybean-cake flour 2.0～5.0;
Semen Maydis powder 1.0～4.0;
Corn steep liquor 0.4～1.2;
Yeast soaks powder 0.02～0.1;
Lime carbonate 0.1～0.3;
Sodium-chlor 0.1～0.5;
Ammonium sulfate 0.05～1.2;
Zinc sulfate 0.01～0.06;
Sal epsom 0.01～0.06;
All the other content are water, sterilize 30 minutes standby;
Culture condition: in seeding tank, cultivated 40 hours～50 hours; Its temperature is controlled at 26 ℃～30 ℃, pH value 6.0～8.0, and tank pressure is controlled at 0.05～0.2Mpa;
(B) fermentation microbiotic generative process: the nutrient solution that forms in the A step seeding tank is sent into fermentor tank ferment; Main following substratum, the percentage composition meter by weight of adding in fermentor tank:
Groundnut meal 2.0～4.0;
Soybean-cake flour 2.0～5.0;
Semen Maydis powder 1.0～4.0;
Corn steep liquor 0.2～1.0;
Yeast soaks powder 0.1～0.5;
Lime carbonate 0.1～0.3;
Sodium-chlor 0.1～0.5;
Ammonium sulfate 0.05～0.2;
Zinc sulfate 0.01～0.06;
All the other content are water, sterilize 30 minutes standby;
After above-mentioned material is put into fermentor tank, adjust the pH value before the sterilization and make it be controlled at 6.0～8.0; Tank pressure is controlled at 0.05～0.2MPa during cultivation, air flow quantity: 1: (0.2～1.0), culture temperature are 26～30 ℃, and the time is 40 hours～50 hours;
(C) fermentation aftertreatment, carry out successively according to the following steps:
1) put jar before, pipeline carries out disinfection in advance;
2) fermented liquid is heated make its temperature reach 80～100 ℃, kept 10 minutes, afterwards cooling rapidly;
3) be that the pH value that the hydrochloric acid of 6N is adjusted fermented liquid makes it be controlled at 3.5～5.0 with concentration; Again fermented liquid is filtered, filter clear liquid;
4) clear liquid is pumped in the concentration tank, in 700mmHg vacuum tightness, vapour pressure 0.02～0.06MPa, temperature is carried out concentrating under reduced pressure under 60 ℃～80 ℃ conditions, concentrated solution is tired be controlled at 15000 units/more than the ml;
5) get finished product behind adding stablizer, the auxiliary agent.
A upward jar inoculum size of being sent into fermentor tank by seeding tank is 8%～15% (V/V), and bacterial strain is 40～50 hours at the incubation time of seeding tank.
All add Semen Maydis oil 0.5～0.8/ ten thousand (V/V) and silicone defoamer 0.5～0.8/ ten thousand (V/V) in seeding tank and the fermentor tank.
In the fermentation post-processing step, stablizer adopts phenylformic acid and copper sulfate, and auxiliary agent is Toween80; Percentage composition meter phenylformic acid adds 0.1%～0.4% of concentrated solution by weight, and copper sulfate is 1%～2% of concentrated solution, and Toween80 percentage composition meter by weight is 0.1%～0.5% of a concentrated solution.
This compound is as the fungal disease of control and treatment plant.
3, advantage and effect: the present invention has stable raw material sources, and cost is low, is fit to large-scale industrial production, mainly contains W-Gum, groundnut meal, yeast powder, glucose etc. as the raw material that is adopted in the fermentating formula.Constant product quality.When guaranteeing that raw material is stable, can control the quality of finished product by the every fermentation parameter of strict control, thus the complete quality system of formation science, this product is stable to light, heat, acid etc., and concentrated solution is preserved at normal temperatures and was not lost efficacy in 2 years.This product low toxicity to the people, animal toxicological harmless, does not have poisoning to farm crop, not skin irritation, eye etc., resistance of rainwater washing against.Control fungal diseases of plants effect is obvious.According to ground vegetables, tobacco, seeding corn and other crops field experiments such as several cities, Liaoning Province, county and Beijing, Jilin, Heilungkiang.
Four, embodiment: the present invention adopts following scheme implementation:
Embodiment 1:
A kind of bacterial strain that produces mycocidal agricultural antibiotic, its formal name used at school are kabicidin streptomycete Streptomyces fungicidicus Yh06, and its culture presevation is numbered CGMCC NO.1247; This strain cell wall belongs to amino acid I type, and its full cell carbohydrate type belongs to sugared type C.This bacterial strain is well-grown on Gause I, No. one, Ke Shi, PDA, glucose yeast agar, Wa Shi nutrient agar substratum; Aerial hyphae and sorus are white in color at early growth period on the Gause I substratum, grizzle or light gray gradually afterwards, and it is yellow or light yellow that substrate mycelium mostly is, soluble pigment yellow, light yellow; Aerial hyphae is gentle straight, terminal slight curving, and fibrillae of spores is chain; Spore ball-type or oval, smooth surface; This bacterial strain gelatine liquefication is stronger, milk is solidified and peptonize, can on Mierocrystalline cellulose, grow but do not decompose, hydrolyzed starch, do not produce class melanochrome and hydrogen sulfide, carbon sources such as glucose, fructose, sucrose, rhamnosyl, lactose, maltose, pectinose, starch, Trisodium Citrate and inositol can be utilized, raffinose can not be utilized.
Adopt this bacterial strain to produce a kind of agricultural antibiotic, its chemical structural formula is:
Its molecular formula is: C
35H
53NO
13, its molecular weight is 695, microbiotic belongs to the tetraenes microbiotic.
The zymotechnique of above-mentioned antifungal agricultural antibiotics is characterized in that: can carry out successively as follows:
(A) bacterial classification expands numerous process:
Adopt kabicidin streptomycete (Streptomyces fungicidicus Yh06), in seeding tank, add following basis and cultivate composition, percentage composition meter by weight:
Groundnut meal 4.0;
Soybean-cake flour 2.0;
Semen Maydis powder 2.0;
Corn steep liquor 0.4;
Yeast soaks powder 0.03;
Lime carbonate 0.15;
Sodium-chlor 0.15;
Ammonium sulfate 0.05;
Zinc sulfate 0.01;
Sal epsom 0.01;
All the other content are water, sterilize 30 minutes.
Culture condition: in seeding tank, cultivated 30 hours～40 hours; Its temperature is controlled at 26 ℃～30 ℃, pH value 6.0～7.0, and tank pressure is controlled at 0.05～0.2MPa,
(B) fermentation microbiotic generative process: the seed culture fluid that the A step forms is sent into fermentor tank, and last jar of inoculum size is 10%～15% (V/V); Main following substratum, the percentage composition meter by weight of adding in fermentor tank:
Groundnut meal 4.0
Soybean-cake flour 3.0;
Semen Maydis powder 1.0;
Corn steep liquor 0.2
Yeast soaks powder 0.15;
Lime carbonate 0.2;
Sodium-chlor 0.2;
Ammonium sulfate 0.05;
Zinc sulfate 0.06
All the other content are water, sterilize 30 minutes standby.
After above-mentioned material is put into fermentor tank, adjust the pH value before the sterilization and make it be controlled at 6.0; Tank pressure is controlled at 0.05MPa during cultivation, air flow quantity: 1: 0.5, culture temperature was 28 ℃, and the time is 45 hours, in the fermentor tank with Semen Maydis oil 0.5/ ten thousand (V/V) and silicone defoamer 0.5/ ten thousand (V/V).
(C) fermentation aftertreatment, carry out successively according to the following steps:
1) put jar before, pipeline carries out disinfection in advance;
2) fermented liquid is heated make its temperature reach 100 ℃, kept 10 minutes, afterwards cooling rapidly;
3) be that the pH value that the hydrochloric acid of 6N is adjusted fermented liquid makes it be controlled at 3.5 with concentration; Again fermented liquid is filtered, filter clear liquid;
4) clear liquid is pumped in the concentration tank, in 700mmHg vacuum tightness, vapour pressure 0.02MPa, temperature is carried out concentrating under reduced pressure under 65 ℃ of conditions, concentrated solution is tired be controlled at 15000 units/more than the ml;
5) get finished product behind adding stablizer, the auxiliary agent.Stablizer adopts phenylformic acid and copper sulfate, and auxiliary agent is Toween80.Percentage composition meter phenylformic acid adds 0.2% of concentrated solution by weight, and copper sulfate is 2% of concentrated solution, and Toween80 percentage composition meter by weight is 0.1% of a concentrated solution.
Embodiment 2,
Experimental field be chosen in the peasant household of Dongling District Ying Da township, Shenyang City production peasant household protection ground tomato.This peasant household produces the history in existing 7 years of tomato, and the leaf mold of early spring, gray mold, early blight etc. have become the regular incidence evil.Test was carried out 2 times every year in 2003 and 2004.Adopt antifungal agricultural antibiotics of the present invention, do not have the generation of above-mentioned disease in these 2 years substantially.In addition, in the experimental plot in several places such as Liaoning, Shandong, implement, find that this antifungal agricultural antibiotics also has the effect that cucumber downy mildew, blight, capsicum epidemic disease, grapevine anthracnose etc. is all had treatment and prevention, illustrate its have wide spectrum, press down mycocidal effect.
Claims (7)
1, a kind of bacterial strain that produces agricultural antibiotic, its formal name used at school are kabicidin streptomycete Streptomycesfungicidicus Yh06, and its culture presevation is numbered CGMCC NO.1247; This strain cell wall belongs to amino acid I type, and its full cell carbohydrate type belongs to sugared type C.
2, a kind of agricultural antibiotic is characterized in that: its chemical structural formula is:
Its molecular formula is C
35H
53NO
13, its molecular weight is 695.
3, a kind of preparation technology of agricultural antibiotic as claimed in claim 2 is characterized in that: this technology is carried out as follows successively:
(A) bacterial classification expands numerous process: adopt kabicidin streptomycete Streptomyces fungicidicusYh06, add following basis and cultivate composition, percentage composition meter by weight in seeding tank:
Groundnut meal 2.0～4.0;
Soybean-cake flour 2.0～5.0;
Semen Maydis powder 1.0～4.0;
Corn steep liquor 0.4～1.2;
Yeast soaks powder 0.02～0.1;
Lime carbonate 0.1～0.3;
Sodium-chlor 0.1～0.5;
Ammonium sulfate 0.05～1.2;
Zinc sulfate 0.01～0.06;
Sal epsom 0.01～0.06;
All the other content are water, sterilize 30 minutes standby;
Culture condition: in seeding tank, cultivated 40 hours～50 hours; Its temperature is controlled at 26 ℃～30 ℃, pH value 6.0～8.0, and tank pressure is controlled at 0.05～0.2Mpa;
(B) fermentation microbiotic generative process: the nutrient solution that forms in the A step seeding tank is sent into fermentor tank ferment; Main following substratum, the percentage composition meter by weight of adding in fermentor tank:
Groundnut meal 2.0～4.0;
Soybean-cake flour 2.0～5.0;
Semen Maydis powder 1.0～4.0;
Corn steep liquor 0.2～1.0;
Yeast soaks powder 0.1～0.5;
Lime carbonate 0.1～0.3;
Sodium-chlor 0.1～0.5;
Ammonium sulfate 0.05～0.2;
Zinc sulfate 0.01～0.06;
All the other content are water, sterilize 30 minutes standby;
After above-mentioned material is put into fermentor tank, adjust the pH value before the sterilization and make it be controlled at 6.0～8.0; Tank pressure is controlled at 0.05～0.2MPa during cultivation, air flow quantity: 1: (0.2～1.0), culture temperature are 26～30C, and the time is 40 hours～50 hours;
(C) fermentation aftertreatment, carry out successively according to the following steps:
1) put jar before, pipeline carries out disinfection in advance;
2) fermented liquid is heated make its temperature reach 80～100 ℃, kept 10 minutes, afterwards cooling rapidly;
3) be that the pH value that the hydrochloric acid of 6N is adjusted fermented liquid makes it be controlled at 3.5～5.0 with concentration; Again fermented liquid is filtered, filter clear liquid;
4) clear liquid is pumped in the concentration tank, in 700mmHg vacuum tightness, vapour pressure 0.02～0.06MPa, temperature is carried out concentrating under reduced pressure under 60 ℃～80 ℃ conditions, concentrated solution is tired be controlled at 15000 units/more than the ml;
5) get finished product behind adding stablizer, the auxiliary agent.
4, the preparation technology of agricultural antibiotic according to claim 3 is characterized in that: a upward jar inoculum size of being sent into fermentor tank by seeding tank is 8%～15% (V/V), and bacterial strain is 40～50 hours at the incubation time of seeding tank.
5, the zymotechnique of agricultural antibiotic according to claim 3 is characterized in that: all add Semen Maydis oil 0.5～0.8/ ten thousand (V/V) and silicone defoamer 0.5～0.8/ ten thousand (V/V) in seeding tank and the fermentor tank.
6, the zymotechnique of the agricultural antibiotic of killing plant pathogenic fungi according to claim 3 is characterized in that: in the fermentation post-processing step, stablizer adopts phenylformic acid and copper sulfate, and auxiliary agent is Toween80; Percentage composition meter phenylformic acid adds 0.1%～0.4% of concentrated solution by weight, and copper sulfate is 1%～2% of concentrated solution, and Toween80 percentage composition meter by weight is 0.1%～0.5% of a concentrated solution.
7, a kind of purposes of agricultural antibiotic as claimed in claim 2 is characterized in that: this compound is as the fungal disease of control and treatment plant.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102533614A (en) * | 2012-02-21 | 2012-07-04 | 浙江大学 | Streptomyces strain and application thereof |
| CN103667114A (en) * | 2013-11-14 | 2014-03-26 | 西北农林科技大学 | Strain capable of controlling ulcer disease of actinidia and preparation method thereof |
| CN106148459A (en) * | 2015-04-27 | 2016-11-23 | 牡丹江佰佳信生物科技有限公司 | The fermentation medium of a kind of Nikemycin and fermentation process |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1260351C (en) * | 2003-04-29 | 2006-06-21 | 云南省微生物研究所 | Streptomyces antifungus |
-
2005
- 2005-03-16 CN CNB2005100460294A patent/CN100422310C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102533614A (en) * | 2012-02-21 | 2012-07-04 | 浙江大学 | Streptomyces strain and application thereof |
| CN103667114A (en) * | 2013-11-14 | 2014-03-26 | 西北农林科技大学 | Strain capable of controlling ulcer disease of actinidia and preparation method thereof |
| CN106148459A (en) * | 2015-04-27 | 2016-11-23 | 牡丹江佰佳信生物科技有限公司 | The fermentation medium of a kind of Nikemycin and fermentation process |
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| CN100422310C (en) | 2008-10-01 |
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Granted publication date: 20081001 Termination date: 20190316 |