CN102268380B - Nodulisporium sp. strain capable of producing high-yield cellulase and xylanase and application thereof - Google Patents
Nodulisporium sp. strain capable of producing high-yield cellulase and xylanase and application thereof Download PDFInfo
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Abstract
The invention discloses a Nodulisporium sp. strain capable of producing high-yield cellulase and xylanase and application thereof. The Nodulisporium sp. strain is Nodulisporium sp. CTB-1, and the registered number of the Nodulisporium sp. strain in Common Microorganism Center of Committee for Culture Collection of Microorganisms is CGMCC No.2482. When the Nodulisporium sp. CTB-1 CGMCC No.2482 is used for preparing an enzyme preparation, the active ingredient of the enzyme preparation is any one of the following: xylanase, cellulase, pectinase and beta-glucanase. The enzyme product prepared from the Nodulisporium sp. CTB-1 can be widely used in textile, feed, cellulosic ethanol, plant extract and other industries.
Description
Technical field
The present invention relates to more piece spore bacterium and the application thereof of a plant height cellulase-producing and zytase.
Background technology
Cellulase is the general name that degraded cellulose generates one group of enzyme of cell-oligosaccharide, cellobiose and glucose, and cellulase almost can be used for all processing industries take plant and plant kernel as raw material, and Application Areas is very extensive.The at present application and development of cellulase is just extending to rapidly many fields such as extraction, papermaking, oil production, fuel alcohol of weaving, food, feed processing, effective ingredients in plant with unprecedented speed.
Xylan is a kind of poly five-carbon sugar, and main component is the D-wood sugar, is the important component part of plant hemicellulose.Zytase is a class with the enzyme system of β-Isosorbide-5-Nitrae-wood sugar glycosidic bond in inscribe and the circumscribed mode degradation of xylan molecule, and this enzyme system also can be widely used in the fields such as weaving, papermaking, food, fodder additives, fuel alcohol.Utilizing the vegetable fibre raw material to produce in the field of alcohol, zytase finally is decomposed into wood sugar to hemicellulose, so that the yeast of assimilation wood sugar is converted into alcohol to wood sugar, increases the ethanol output capacity of cellulose raw material.Therefore, be the industrial circle of raw material in many vegetable fibre matter of utilizing, all need simultaneously the effect of cellulase and zytase, and the industrial production of zymin is still produced single enzyme system at present.
The amount of the waste water of China's agricultural wastes and paper industry discharging is all very large, only the stalk annual production just reaches 700,000,000 tons, these agricultural wastes are incinerated by the peasant mostly, and contain a large amount of recycling paper wastes in the paper waste of discharging, this is serious environment pollution not only, and has wasted precious resources.
The more piece spore has the applied research of bibliographical information few before belonging to eubacteriales, and its purposes mostly is the fermentative production taxol, does not also have the report for the production of cellulase and zytase.
Summary of the invention
The purpose of this invention is to provide more piece spore bacterium and the application thereof of a plant height cellulase-producing and zytase.
More piece spore bacterium provided by the present invention is more piece spore (Nodulisporium sp.) CTB-1, this bacterial strain is that the bacterial strain that obtains with screening and separating from the soil sample in one farmland, suburbs, Wuxi is as original strain, obtain through mutagenic and breeding repeatedly, through being accredited as more piece spore (Nodulisporium sp.).More piece spore (Nodulisporium sp.) CTB-1 bacterial strain was preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center and (is called for short CGMCC on 05 07th, 2008, the address is: the Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), its registration is numbered CGMCC No.2482.
More piece spore provided by the invention (Nodulisporium sp.) CTB-1 CGMCC No.2482 has following microbial characteristic:
Be 60-70mm at 25 ℃ of colony diameters after cultivating 7 days on the PDA flat board.Initial stage is white hypha, and the later stage can form the green spore district that produces.It is faint yellow that substratum gradually is.Conidiophore mycelia shape or doleiform, long 7.5-30 μ m, diameter 2-4 μ m, obvious scar is arranged at the conidiogenous cell top.Conidium is avette, oval to cylindricality, and 3.0-7.5 μ m * 2.0-3.5 μ m is unicellular, filbert, smooth.Give birth to or a life on the chlamydospore top, subsphaeroidal, diameter 5-12 μ m.
More piece spore provided by the present invention (Nodulisporium sp.) CTB-1 CGMCC No.2482 can be used for preparing zymin.
The activeconstituents of described zymin is any one enzyme in following four kinds of enzymes: zytase, cellulase, polygalacturonase and beta-glucanase;
Described preparation zymin comprises the steps: that more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482 is carried out liquid fermenting collects fermented liquid, extracts the activeconstituents of described zymin from described fermented liquid.
In the described preparation zymin, can adopt existing zytase extracting method, cellulase extracting method, polygalacturonase extracting method or beta-glucanase extracting method to extract corresponding enzyme.
The substratum that described fermentation is used is comprised of water and following component:
1) glucose;
2) agriculture and industry waste of at least a cellulose;
3) be selected from least a raw material in following six kinds of raw materials: dregs of beans, cotton dregs, peanut meal, yeast powder, corn steep liquor and wheat bran;
4) nutritive salt;
5) trace element;
The pH=5.0 of described substratum ± 0.2.
The concentration of described each component in described substratum is as follows: described glucose 2g/L-10g/L specifically can be 2g/L-4g/L; The agriculture and industry waste total amount 5g/L-50g/L of described cellulose specifically can be 25g/L-40g/L; Described raw material total amount 5g/L-60g/L specifically can be 13g/L-32g/L; Described nutritive salt is by CaCl
2, urea, KH
2PO
4, (NH
4)
2SO
4And MgSO
4Form, in the described substratum, described CaCl
2Concentration be that the concentration of 0.6g/L, described urea is 0.6g/L, described KH
2PO
4Concentration be 4g/L, described (NH
4)
2SO
4Concentration be 2.8g/L and described MgSO
4Concentration be 0.6g/L; Described trace element is by FeSO
4, MnSO
4, ZnSO
4And CoCl
2Form FeSO described in the described substratum
4Concentration be 0.01g/L, described MnSO
4Concentration be 0.0032g/L, described ZnSO
4Concentration be 0.0028g/L and described CoCl
2Concentration be 0.0074g/L.
The agriculture and industry waste of described cellulose is rice straw powder, wheat straw powder, corn straw powder, beet pulp, wood sawdust, paper waste and maize peel.
The temperature of described fermentation is 33 ℃.
The present invention adopts agriculture and industry waste to pass through liquid fermentation and culture more piece spore (Nodulisporium sp.) CTB-1CGMCC No.2482, these four kinds of enzymes of production of cellulose enzyme, zytase, beta-glucanase and polygalacturonase, and the method has the following advantages:
1) these four kinds of enzymes of cellulase, zytase, beta-glucanase and polygalacturonase of producing can be monose with Degrading plant biomass;
2) with the main ingredient of agriculture and industry waste as fermention medium, and liquid fermentation production can directly prepare prozyme, and the production cost of zymin is reduced, and application performance improves, and also can promote simultaneously the utilization of vegetable fibre raw material, environmental contamination reduction;
3) adopt filter method to remove fermented liquid solid residue and thalline, guaranteed that enzyme fluidity matter is not subjected to the interference of these materials; Be further purified zymin by ultrafiltration and concentration, reduced drying cost; By spraying drying, organized enzyme is reclaimed;
4) enzyme product of the present invention's generation is applied in textile industry, and the front and back arrangement of cotton, linen is carried out with bathing, and shortens technological process, enhances productivity; Being applied in feedstuff industry and producting stalk fodder processing aspect can significantly improve feed and tire, decompose the anti-dietetic alimentation factor that contains in the feed, promote animal to the digestibility of nutrition; Be applied in the production aspect of cellulosic ethanol, can improve the vegetable fibre raw material utilization ratio, greatly reduce production costs, reduce lumber consumption, protection vegetation, save energy.
Embodiment
Employed experimental technique is ordinary method if no special instructions among the following embodiment.
Used material, reagent etc. if no special instructions, all can obtain from commercial channels among the following embodiment.
The raw material sources that adopt in the fermention medium among the following embodiment are as follows:
Wheat straw powder: wheat stalk crushed material, particle diameter 40 orders.
Corn stalk powder: corn stalk powder minces, particle diameter 40 orders.
Rice straw powder: rice straw powder minces, particle diameter 40 orders.
Yeast powder: the food-drink fermentation industry by-products such as beer.
Peanut meal: be the Semen arachidis hypogaeae that shells be raw material, the by product after oil is got in squeezing or lixiviate.Peanut meal is available from Shandong Jade Emperor oil and foodstuffs company limited among the following embodiment.
Dregs of beans: be a kind of byproduct that obtains behind the soybean extracting bean oil.Dregs of beans is available from Hebei Sanhe Hopefull Grain ﹠ Oil Group Co., Ltd. among the following embodiment.
Wheat bran: after the outermost epidermis of wheat, wheat are processed by fluffing machine, become flour and wheat bran two portions, wheat bran is exactly the crust of wheat.The scope of wheat bran nitrogen content is at 10-30%.
Maize peel: be the crushed material of the outer bract of corn ear, particle diameter 40 orders.
Corn steep liquor: be the by product of W-Gum processed, be about to corn grain and soak with sulfurous acid first that soak solution concentrates and makes filemot liquid is corn steep liquor.Corn steep liquor is available from continent, Shandong biotechnology (Shandong) company limited among the following embodiment.
Paper waste: the cellulosic material that the waste paper such as newspaper are made through techniques such as pulverizing, purification, floatation and ink removing, dehydration, dryings.
The nutritive salt of mentioning among the following embodiment, trace element are comprised of following material:
Nutritive salt: by CaCl
2, urea, KH
2PO
4, (NH
4)
2SO
4And MgSO
4Form;
Trace element: by FeSO
4, MnSO
4, ZnSO
4And CoCl
2Form.
Among the following embodiment, the prescription of employed seed slant medium and seed tank culture base is as follows:
Seed slant medium: Xylo-Mucine (CMC-Na) 15g/L, peptone 5g/L, agar 15g/L, CaCl
20.3g/L, urea 0.3g/L, KH
2PO
42g/L, (NH
4)
2SO
41.4g/L, MgSO
40.3g/L, FeSO
40.005g/L, MnSO
40.0016g/L, ZnSO
40.0014g/L, CoCl
20.0037g/L take water as solvent, the pH value is 6.0 ± 0.1.
Seed tank culture base: glucose 10g/L, peptone 5g/L, CaCl
20.3g/L, urea 0.3g/L, KH
2PO
42g/L, (NH
4)
2SO
41.4g/L, MgSO
40.3g/L, FeSO
40.025g/L, MnSO
40.008g/L, ZnSO
40.007g/L, CoCl
20.0185g/L, citric acid 5.7g/L, Trisodium Citrate 6.73g/L, take water as solvent, the pH value is 5.0 ± 0.2.
Among the following embodiment, related enzyme work is defined as follows:
Cellulase (CMC) vigor: adopt the DNS method, at pH=4.8, under 50 ℃ of conditions, 1ml enzyme liquid catalysis 2% Xylo-Mucine decomposes, per hour produce 1.0mg reducing sugar (with glucose meter) and be defined as a unit of activity, represent with U/mL or U/g.
Xylanase activity: adopt the DNS method, at pH=4.8, under 50 ℃ of conditions, 1ml enzyme liquid catalysis 0.5% xylan decomposes, and per hour produces 1.0mg reducing sugar (in wood sugar) and is defined as a unit of activity, represents with U/mL or U/g.
Polygalacturonase qualitative detection: make spore suspension, coat the flat board take pectin as sole carbon source after the dilution suitable multiple, place 28 ℃ constant incubator to cultivate 3 days, take out, spray is with 0.005% coeruleum bromocresolis solution, can see significantly that yellow look becomes circle (pectin generates free pectic acid after by the pectin enzymic hydrolysis and is acid and makes indicator discoloration), illustrates that this more piece spore bacterial strain can produce polygalacturonase, and decomposes and utilize pectin.
Beta-glucanase qualitative detection: make spore suspension, coat the Congo red nutrition of beta-glucan dull and stereotyped (GCN is dull and stereotyped) after the dilution suitable multiple, place 28 ℃ constant incubator to cultivate 3 days, can see obvious transparent circle (beta-glucan be hydrolyzed into the small molecules such as glucose can not with Congo red aobvious redness), illustrate that this more piece spore bacterial strain can produce beta-glucanase.
Separation, purifying and the evaluation of embodiment 1, more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482
1, the separation of more piece spore (Nodulisporium sp.)
Get the lower soil in farmland, one place, suburbs, Wuxi, be diluted to suitable multiple with stroke-physiological saline solution, be coated with flat board, select substratum respectively take Xylo-Mucine (CMC-Na) and xylan as sole carbon source, filter out the bacterium colony that to grow at the flat board of two kinds of single carbon sources simultaneously with photolithography, pick out 12 strain bacterium purifying preservations; With congo red staining transparent circle method, select 5 larger strain bacterium of transparent circle and colony diameter, sieve again all relatively high bacterium C1 of the strain cellulase-producing vigor that obtains and Xylanase activity through shaking flask.
2, the mutagenic and breeding of more piece spore (Nodulisporium sp.)
As the bacterium that sets out, through ultraviolet ray, nitrosoguanidine multiple mutated, obtain mutagenic fungi CTB-1 with bacterial strain C1.Concrete grammar is as follows:
With normal saline flushing starting strain C1 inclined-plane, after the absorbent cotton filtration, be mixed with 10
6The spore suspension of individual/ml places spore suspension under the UV-light (30W, 30cm), and 4min is stirred in the darkroom.Then in this spore suspension, add the 1mg/mL nitrosoguanidine, 33 ℃ of waters bath with thermostatic control, mutagenesis 40min.
Carry out the plate separation screening after the mutagenesis, select 20-30 single bacterium colony, do the shake flask fermentation test, measure and produce enzyme activity, take the product enzyme activity of zytase and cellulase as the main index that detects, select 2～3 strain enzymes to live high bacterial strain as the superior strain revision test, select the highest bacterial strain of a strain output as the starting strain of next time mutagenesis, through obtaining the bacterial strain that a plant height produces zytase and cellulase after the several times mutagenesis, with this bacterial strain called after CTB-1.
Bacterial strain CTB-1 is accredited as more piece spore (Nodulisporium sp.) through China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC), registers on the books and be numbered CGMCC No.2482 in the preservation center.Its biological characteristics is as follows:
On the PDA flat board 25 ℃ cultivate 7 days after colony diameter 60-70mm.Initial stage is white hypha, and the later stage can form the green spore district that produces.It is faint yellow that substratum gradually is.Conidiophore mycelia shape or doleiform, long 7.5-30 μ m, diameter 2-4 μ m, obvious scar is arranged at the conidiogenous cell top.Conidium is avette, oval to cylindricality, and 3.0-7.5 μ m * 2.0-3.5 μ m is unicellular, filbert, smooth.Give birth to or a life on the chlamydospore top, subsphaeroidal, diameter 5-12 μ m.
Embodiment 2, more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482 utilize agriculture and industry waste simultaneously High Cellulase Production and zytase prozyme and enzyme activity determination thereof
1, on more piece spore (Nodulisporium sp.) the CTB-1 CGMCC No.2482 access seed slant medium with embodiment 1, after 33 ℃ of thermostat containers are cultivated 5 days, uses the stroke-physiological saline solution wash-out, be made into bacteria suspension.
2, bacteria suspension changes in the complete seeding tank of the sterilization that feeds intake, and stirs aerated culture in 33 ℃ and obtains seed liquor in 36 hours.
3, seed liquor is transferred to the complete fermentor tank (fermention medium: wheat straw powder 10g/L, corn stalk powder 15g/L, yeast powder 2g/L, peanut meal 10g/L, wheat bran 3g/L, glucose 2g/L, CaCl of sterilization that feeds intake through seed transferring pipeline
20.6g/L, urea 0.6g/L, KH
2PO
44g/L, (NH
4)
2SO
42.8g/L, MgSO
40.6g/L, FeSO
40.01g/L, MnSO
40.0032g/L, ZnSO
40.0028g/L, CoCl
20.0074g/L take water as solvent, the pH value is 5.0 ± 0.2) in, in 33 ℃ of stirring ventilations, cultivated 160 hours, obtaining the cellulose enzyme activity is that 1896U/ml and Xylanase activity are the fermented liquid of 3309U/ml.
4, fermented liquid is put into the flocculation tank through the blowing pipeline, in the flocculation tank, add 20g/L super-cell (700 orders, Inner Mongol spring bio-diatomite limited liability company), through the plate-and-frame filter press press filtration, clear liquid is squeezed into the constant temperature container for storing liquid and is carried out 8 times of ultrafiltration and concentration through 5000 molecular weight ultrafiltration apparatuss and obtain enzyme liquid work in-process.
5, enzyme liquid work in-process add the 100g/L maltodextrin, be that 170 ℃, temperature out are that 70 ℃ spraying drying is made the solid complex enzyme preparation through temperature in, its cellulase activity reaches 10.6 ten thousand U/g, Xylanase activity reaches 19.4 ten thousand U/g, through qualitative detection, also have polygalacturonase and 1,4 beta-glucanase activity.
Embodiment 3, more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482 utilize agriculture and industry waste simultaneously High Cellulase Production and zytase prozyme and enzyme activity determination thereof
1, on more piece spore (Nodulisporium sp.) the CTB-1 CGMCC No.2482 access seed slant medium with embodiment 1, after 33 ℃ of thermostat containers are cultivated 5 days, uses the stroke-physiological saline solution wash-out, be made into bacteria suspension.
2, bacteria suspension changes in the complete seeding tank of the sterilization that feeds intake, and stirs aerated culture in 33 ℃ and obtains seed liquor in 36 hours.
3, seed liquor is transferred to the complete fermentor tank (fermention medium: rice straw powder 15g/L, maize peel 25g/L, dregs of beans 10g/L, wheat bran 3g/L, glucose 2g/L, CaCl of sterilization that feeds intake through seed transferring pipeline
20.6g/L, urea 0.6g/L, KH
2PO
44g/L, (NH
4)
2SO
42.8g/L, MgSO
40.6g/L, FeSO
40.01g/L, MnSO
40.0032g/L, ZnSO
40.0028g/L, CoCl
20.0074g/L take water as solvent, the pH value is 5.0 ± 0.2) in, in 33 ℃ of stirring ventilations, cultivated 160 hours, obtaining the cellulose enzyme activity is that 1983U/ml and Xylanase activity are the fermented liquid of 3420U/ml.
4, fermented liquid is put into the flocculation tank through the blowing pipeline, in the flocculation tank, add 20g/L super-cell (700 orders, Inner Mongol spring bio-diatomite limited liability company), through the plate-and-frame filter press press filtration, clear liquid is squeezed into the constant temperature container for storing liquid and is carried out 8 times of ultrafiltration and concentration through 5000 molecular weight ultrafiltration apparatuss and obtain enzyme liquid work in-process.
5, add the 100g/L maltodextrin in the enzyme liquid work in-process, be that 170 ℃, temperature out are that 70 ℃ spraying drying is made solid complex enzyme through temperature in, its cellulase activity reaches 11.8 ten thousand U/g, Xylanase activity reaches 200,000 U/g, through qualitative detection, also contain polygalacturonase and 1,4 beta-glucanase activity.
Embodiment 4, more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482 utilize agriculture and industry waste simultaneously High Cellulase Production and zytase prozyme and enzyme activity determination thereof
1, on more piece spore (Nodulisporium sp.) the CTB-1 CGMCC No.2482 access seed slant medium with embodiment 1, after 33 ℃ of thermostat containers are cultivated 5 days, uses the stroke-physiological saline solution wash-out, be made into bacteria suspension.
2, bacteria suspension changes in the complete seeding tank of the sterilization that feeds intake, and stirs aerated culture in 33 ℃ and obtains seed liquor in 36 hours.
3, seed liquor is transferred to the complete fermentor tank (fermention medium: paper waste 15g/L, corn steep liquor 20g/L, maize peel 25g/L, wheat bran 3g/L, glucose 2g/L, CaCl of sterilization that feeds intake through seed transferring pipeline
20.6g/L, urea 0.6g/L, KH
2PO
44g/L, (NH
4)
2SO
42.8g/L, MgSO
40.6g/L, FeSO
40.01g/L, MnSO
40.0032g/L, ZnSO
40.0028g/L, CoCl
20.0074g/L take water as solvent, the pH value is 5.0 ± 0.2) in, in 33 ℃ of stirring ventilations, cultivated 160 hours, obtaining the cellulose enzyme activity is that 2140U/ml and Xylanase activity are the fermented liquid of 3554U/ml.
4, fermented liquid is put into the flocculation tank through the blowing pipeline, in the flocculation tank, add 20g/L super-cell (700 orders, Inner Mongol spring bio-diatomite limited liability company), through the plate-and-frame filter press press filtration, clear liquid is squeezed into the constant temperature container for storing liquid and is carried out 8 times of ultrafiltration and concentration through 5000 molecular weight ultrafiltration apparatuss and obtain enzyme liquid work in-process.
5, add the 100g/L maltodextrin in the enzyme liquid work in-process, be that 170 ℃, temperature out are that 70 ℃ spraying drying is made solid complex enzyme through temperature in, its cellulase activity reaches 120,000 U/g, Xylanase activity reaches 210,000 U/g, through qualitative detection, also contain polygalacturonase and 1,4 beta-glucanase activity.
Embodiment 5, more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482 utilize agriculture and industry waste simultaneously High Cellulase Production and zytase prozyme and enzyme activity determination thereof
1, on more piece spore (Nodulisporium sp.) the CTB-1 CGMCC No.2482 access seed slant medium with embodiment 1, after 33 ℃ of thermostat containers are cultivated 5 days, uses the stroke-physiological saline solution wash-out, be made into bacteria suspension.
2, bacteria suspension changes in the complete seeding tank of the sterilization that feeds intake, and stirs aerated culture in 33 ℃ and obtains seed liquor in 36 hours.
3, seed liquor is transferred to the complete fermentor tank (fermention medium: rice straw powder 10g/L, corn stalk powder 15g/L, yeast powder 2g/L, dregs of beans 10g/L, corn steep liquor 10g/L, wheat bran 10g/L, glucose 4g/L, CaCl of sterilization that feeds intake through seed transferring pipeline
20.6g/L, urea 0.6g/L, KH
2PO
44g/L, (NH
4)
2SO
42.8g/L, MgSO
40.6g/L, FeSO
40.01g/L, MnSO
40.0032g/L, ZnSO
40.0028g/L, CoCl
20.0074g/L take water as solvent, the pH value is 5.0 ± 0.2) in, in 33 ℃ of stirring ventilations, cultivated 160 hours, obtaining the cellulose enzyme activity is that 1983U/ml and Xylanase activity are the fermented liquid of 3710U/ml.
4, fermented liquid is put into the flocculation tank through the blowing pipeline, in the flocculation tank, add 20g/L super-cell (700 orders, Inner Mongol spring bio-diatomite limited liability company), through the plate-and-frame filter press press filtration, clear liquid is squeezed into the constant temperature container for storing liquid and is carried out 8 times of ultrafiltration and concentration through 5000 molecular weight ultrafiltration apparatuss and obtain enzyme liquid work in-process.
5, add the 100g/L maltodextrin in the enzyme liquid work in-process, be that 170 ℃, temperature out are that 70 ℃ spraying drying is made solid complex enzyme through temperature in, its cellulase activity reaches 11.3 ten thousand U/g, Xylanase activity reaches 220,000 U/g, through qualitative detection, also contain polygalacturonase and 1,4 beta-glucanase activity.
Claims (6)
1. more piece spore (Nodulisporium sp.) CTB-1 is numbered CGMCC No.2482 registering on the books of China Committee for Culture Collection of Microorganisms common micro-organisms center.
2. the application of more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482 in the preparation zymin;
The activeconstituents of described zymin be in following four kinds of enzymes any one: zytase, cellulase, polygalacturonase and beta-glucanase.
3. application according to claim 2, it is characterized in that: the described method for preparing zymin comprises the steps: that more piece spore (Nodulisporium sp.) CTB-1 CGMCC No.2482 is carried out liquid fermenting collects fermented liquid, extracts the activeconstituents of described zymin from described fermented liquid.
4. application according to claim 3 is characterized in that: the substratum that described fermentation is used is comprised of water and following component:
1) glucose;
2) at least a in rice straw powder, wheat straw powder, corn straw powder, beet pulp, wood sawdust, paper waste and the maize peel;
3) be selected from least a raw material in following six kinds of raw materials: dregs of beans, cotton dregs, peanut meal, yeast powder, corn steep liquor and wheat bran;
4) CaCl
2, urea, KH
2PO
4, (NH
4)
2SO
4And MgSO
4
5) FeSO
4, MnSO
4, ZnSO
4And CoCl
2
The pH=5.0 of described substratum ± 0.2.
5. application according to claim 4 is characterized in that: the concentration of described each component in described substratum is as follows: described glucose 2g/L-10g/L; Described 2) the total amount 5g/L-50g/L of component described in; Described 3) 5g/L-60g/L of raw material total amount described in; In the described substratum, described CaCl
2Concentration be that the concentration of 0.6g/L, described urea is 0.6g/L, described KH
2PO
4Concentration be 4g/L, described (NH
4)
2SO
4Concentration be 2.8g/L and described MgSO
4Concentration be 0.6g/L; FeSO described in the described substratum
4Concentration be 0.01g/L, described MnSO
4Concentration be 0.0032g/L, described ZnSO
4Concentration be 0.0028g/L and described CoCl
2Concentration be 0.0074g/L.
6. arbitrary described application according to claim 3-5 is characterized in that: the temperature of described fermentation is 33 ℃.
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| US20060111241A1 (en) * | 2004-11-19 | 2006-05-25 | Gerwick Ben C Iii | Methylidene mevalonates and their use as herbicides |
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2011
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060111241A1 (en) * | 2004-11-19 | 2006-05-25 | Gerwick Ben C Iii | Methylidene mevalonates and their use as herbicides |
Non-Patent Citations (2)
| Title |
|---|
| 刘晓兰等.树状多节孢发酵生产紫杉醇工艺条件的初步研究.《菌物系统》.2002,第21卷(第02期),246-251. * |
| 解玉红等.同工酶技术在树状多节孢研究中的应用.《天津理工大学学报》.2005,第21卷(第03期),38-41. * |
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| CN102268380A (en) | 2011-12-07 |
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