CN104531640A - Aspergillus oryzae fermentation liquor, corn stalk sugar liquor prepared through same, and preparation method and application of corn stalk sugar liquor - Google Patents
Aspergillus oryzae fermentation liquor, corn stalk sugar liquor prepared through same, and preparation method and application of corn stalk sugar liquor Download PDFInfo
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Abstract
The invention provides aspergillus oryzae fermentation liquor and a preparation method of the aspergillus oryzae fermentation liquor. The aspergillus oryzae fermentation liquor contains manganese peroxide with the enzymatic activity being 100-10000U/L, endo-cellulase with the enzymatic activity being 150-3000U/L, beta-dextranase with enzymatic activity being 150-3000U/L, and beta-mannase with enzymatic activity being 50-2000U/L. The invention further provides a process technology of corn stalk sugar liquor. The corn stalk sugar liquor uses corn stalks as main raw materials and is prepared by adding a hydrogen dioxide solution to degrade the corn stalks through the aspergillus oryzae fermentation liquor. The process technology has the advantages of being short in production cycle, small in occupied space, low in production cost and high in sugar yield. In addition, the prepared corn stalk sugar liquor can be used as a carbon source raw material for producing butanol, ethyl alcohol and other fermentation industries.
Description
Technical field
The invention belongs to fermentation engineering and enzyme engineering two large field, particularly a kind of aspergillus oryzae fermented liquid, the maize straw liquid glucose prepared by this fermented liquid and preparation method and purposes.
Background technology
China is large agricultural country, and annual agricultural crop straw annual production exceedes 600,000,000 tons, and except being used as weaving on a small quantity, except paper industry, major part, with the form impouring environment piled up, burn, had both been wasted resource and also caused to environment and greatly endanger.
Exhausted most industrial microorganism is all organotroph (heterotroph), so the prevailing energy is from carbon source, as carbohydrate, lipid and protein.The carbon that cell is basic simultaneously is also derive from carbon source.The carbon source major part used in fermentation industry is from saccharide compound.Saccharide compound main at present, from the starch of plant, therefore causes fermentation industry to there is the problem of striving grain with people.Developing rapidly due to science and technology simultaneously, increasing to the demand of the energy, the current energy mainly fossil energy, a small amount of wind energy and water energy, these energy more and more can not meet the demand of social development, therefore find the problem that the new energy also becomes extremely urgent.
The stalk of plant contains Mierocrystalline cellulose, the hemicellulose of about 25%, the xylogen of about 15% of about 30%, is the source of natural carbohydrate.Especially Mierocrystalline cellulose and hemicellulose, main component is carbohydrate, is fermentable the most natural industrial source.But there is the compound that xylogen etc. is not easily degraded in stalk, mainly because the combination of Mierocrystalline cellulose, hemicellulose and xylogen in stalk: xylogen is combined with covalent linkage form with hemicellulose, cellulosic molecule is embedded in centre, the enzyme of lignin degrading is not easily contacted with cellulosic molecule; Add the water-insoluble of xylogen, complicated chemical structure, also bring great difficulty to degraded.Up to the present stalk is as agricultural wastes, or burned or directly abandon it, both pollutes environment, and wastes resource again.Therefore, how lignin degrading develops the matter of utmost importance that stalk resource becomes agricultural sustainable development.
The method of current degrading straw mainly contains physical treatment, chemical treatment and biological treatment.Physical treatment mainly contains high pressure steam explosion, extruding puffing and microwave-heating.High pressure steam explosion is placed in encloses container by material and water, be heated to certain temperature, pressure about 4.0MPa is kept to arrive several minutes in several seconds, then bust pressure carries out explosion to material, hemicellulose and xylogen articulamentum are destroyed, make Mierocrystalline cellulose expose more active group, can fully contact with Cellulase Molecules and degrade.Extruding puffing is the modified rear input extrusion chamber that added water by stalk, rely in stalk and extrusion chamber mutually extrude between swivel nut wall and screw rod, rubbing effect, generation heat and pressure, after stalk is extruded nozzle, pressure pcl so declines, thus makes the technological operation of stalk enlarged volume.These two kinds of method compositions are higher, need large-scale equipment, and xylogen also needs process; Microwave-heating causes heat effect with non-thermal form, the collision of speeding-up ion and other molecules, rapidly rotating dipole.But this method is also just limited in testing laboratory at present, be difficult to form large-scale strong industry.Method of chemical treatment mainly contains dilute acid pretreatment, alkaline purification and ammonia treatment.Dilute acid pretreatment utilizes sulphuric acid hydrolysis glycosidic link, ester bond etc., and the reticulated structure of corn stalk fiber is loosened, but the method consumes a large amount of organic acids or mineral acid, xylogen still needs further process, is not suitable with large-scale industrial production.Alkaline purification and ammonia treatment utilize xylogen to be dissolved in basic solution, and the same with acid treatment exist problem of environmental pollution equally.Generally all there is the more difficult grasp of reaction conditions in these methods, temperature of reaction is high, and the treatment time is long, and take up room large, treatment effect is undesirable, can not remove the problems such as lignocellulose.Stalk of carrying out a biological disposal upon in recent years is more and more subject to people's attention, and bioremediation mainly comprises ensiling, microbial and enzymolysis.Ensiling is exactly under anaerobic, glucide in feed is decomposed generation lactic acid by the probioticss such as the milk-acid bacteria utilizing stalk itself contained, suppress or kill other various harmful microorganisms, as spoilage organism, mould etc., thus reaching long-term object of preserving feed.The principle of microbial and ensiling is closely similar, just pass through to add a certain amount of microbe additive as stalk fermentation live bacilli, white-rot fungi, yeast etc. before fermentation, then utilize these microorganisms to carry out decomposition to stalk to utilize, stalk is softened, the organic carbon hydrates such as Mierocrystalline cellulose wherein, hemicellulose and xylogen are converted into carbohydrate, finally fermentation becomes lactic acid and some other voltaile fatty acid, thus improves rumen microorganism to the utilization ratio of stalk.But the bioremediation cycle is oversize, occupied ground is comparatively large, and particularly aerobic biodegradation is all the more so.Enzymolysis process is that select can the single or compound enzyme of hydrocellulose, hemicellulose and xylogen, processes meeting under enzyme action condition, thus reduces fibrous substance ratio, improve the method for soluble sugar content to stalk.The key of microbiological deterioration xylogen is utilized to be the enzyme that microorganism can secrete degrading straw.
For biological process produce degrading straw zymin patent, research all less.Patent (application number: object 200610109413) is the weak point overcoming current white rot fungus degrading xylogen, and provides a kind of microbiobacterial agent of novel lignin degrading and utilize the method for above-mentioned microbial inoculum lignocellulose degradation.Microbiobacterial agent is made up of pseudomonas, bacillus amyloliquefaciens, subtilis.Further, biological degradation disclosed in prior art mainly adopts solid-state fermentation technology, limit enzymatic production, limit lignin degrading, but inevitable along with cellulose degradation during the fermentation, makes the finished product sugar yield low; Secondly, large-scale solid state fermentation is all the fermentation mode of open type, and fermentation period is very long, and therefore inevitably living contaminants, the particularly pollution of poisonous and harmful bacterium are difficult to control during the fermentation; Again, the solid state fermentation cycle is in the past oversize, and occupied ground is larger.
Summary of the invention
In order to solve the problems of the technologies described above, the invention provides a kind of new aspergillus oryzae fermented liquid, containing the enzyme needed for microorganism secretion degrading maize straws in this fermented liquid; And the present invention provides with aspergillus oryzae fermented liquid for catalyzer on the other hand, add hydrogen peroxide degradation maize straw, obtain the liquid glucose of degrading maize straws; The liquid glucose preparation method provided is with short production cycle, and occupied ground is little, and production cost is low, and liquid glucose yield is high, is applicable to suitability for industrialized production.
The concrete technical scheme of the present invention is as follows:
The invention provides a kind of aspergillus oryzae fermented liquid, this aspergillus oryzae fermented liquid contains manganese peroxidase enzymic activity 100 ~ 10000U/L, endo cellulase enzymic activity 150 ~ 3000U/L, beta-glucanase enzymic activity 150 ~ 3000U/L, 'beta '-mannase enzymic activity 50 ~ 2000U/L; Described aspergillus oryzae fermented liquid take maize straw as main raw material, and aspergillus oryzae is bacterial strain, obtains through test tube enlarged culturing, liquid submerged culture, seeding tank seed culture and liquid fermentation and culture.
Aspergillus oryzae bacterial classification used in the present invention by united States food and drug administration (FDA), U.S. feed Gong Ding association (AAFCO) and the Ministry of Agriculture of China assert can the aspergillus oryzae bacterial classification of feed level microbe additive of Direct-fed animal; And bacterial classification is in the past Phanerochaete chrysosporium, worm plan wax bacterium and some edible fungus species, bacterium and actinomycetes, except edible mushrooms, its security is all without demonstration.
The present invention's aspergillus oryzae bacterial classification used is any one the aspergillus oryzae bacterial classification through the preservation of preservation center, as the bacterial classification of the preservations such as China General Microbiological culture presevation administrative center (CCGMC), Chinese agriculture Microbiological Culture Collection administrative center (ACCC), Chinese industrial Microbiological Culture Collection administrative center (CICC), Chinese medicine Microbiological Culture Collection administrative center (CMCC), National Veterinary Culture Collection (CVCC) and microbiotic bacterial classification preservation pipe reason center.
Main raw material maize straw used in the present invention is any one maize straw, as the maize straw of conventional corn stalk, sweet corn stalk, field corn stalk, blue or green maize straw and maturation.Preferably, first maize straw through pulverizing, crosses 20-100 mesh sieve.
The present invention provides the preparation method of aspergillus oryzae fermented liquid on the other hand, and the method comprises the steps:
A. test tube enlarged culturing: aspergillus oryzae bacterial classification is inoculated in potato dextrose medium, cultivates, obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by aspergillus oryzae test tube slant spore inoculating in the shaking flask that liquid submerged culture base is housed, cultivate, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by liquid shaking bottle strain inoculation in seed tank culture base, cultivate, obtained first class seed pot bacterial classification;
D. liquid fermentation and culture: by first class seed pot strain inoculation in fermention medium, cultivate, obtained aspergillus oryzae fermented liquid.
Described potato dextrose medium source (Zhu Gejian, Wang Zhengxiang. industrial microorganism experimental technique handbook, 1994:367 page).
Further improvement, the preparation method of described aspergillus oryzae fermented liquid comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 3-8d at temperature is 25-45 DEG C, obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by aspergillus oryzae test tube slant spore inoculating in the shaking flask that liquid submerged culture base is housed, cultivates 24-72h at temperature is 20-45 DEG C, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by liquid shaking bottle strain inoculation in seed tank culture base, at temperature is 20-45 DEG C, cultivates 18-72h, obtained first class seed pot bacterial classification;
D. liquid fermentation and culture: by first class seed pot strain inoculation in fermention medium, at temperature 20-45 DEG C, cultivates 3-10d, obtained aspergillus oryzae fermented liquid.
Further improvement, the weight of every 100mL liquid submerged culture base and each composition contained by every 100mL seed tank culture base is:
Wheat bran 0.2-2g maltose 0.1-3g peptone 0.1-1g
Ferrous sulfate 0.0005-0.01g magnesium sulfate 0.005-0.2g Repone K 0.01-0.2g.
Preferably, the weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 1-8g maltose 0.1-3g wood sugar 0-3g
Peptone 0.3-3g ferrous sulfate 0.005-0.02g magnesium sulfate 0.05-0.2g
Repone K 0.01-1g wheat bran 0.1-5 Semen Maydis powder 0-1g.
The granularity of wheat bran used in the present invention is 10-100 order.
Liquid submerged culture base provided by the present invention and fermention medium are through 100-130 DEG C of sterilizing 30-120min.
Further improvement, the preparation method of described aspergillus oryzae fermented liquid comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 3-8d at temperature is 25-45 DEG C, then in 1-10 DEG C of preservation, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask that 40-150mL liquid submerged culture base is housed, the rotating speed of triangular flask is 60-200 rev/min is that 24-72h cultivated by the shaking table of 20-45 DEG C in temperature, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by inoculum size be 1-20% by liquid shaking bottle strain inoculation in seed tank culture base, be 20-45 DEG C in temperature, mixing speed is under the condition of 50-200 rev/min, cultivates 18-72h, obtained first class seed pot bacterial classification;
Described inoculum size is the volume ratio of liquid shaking bottle bacterial classification and liquid submerged culture base;
D. liquid fermentation and culture: by inoculum size 2-20% by first class seed pot strain inoculation in fermention medium, at temperature 20-45 DEG C, under mixing speed 50-200 rev/min condition, cultivate 3-10d, obtained aspergillus oryzae fermented liquid;
Described inoculum size is the volume ratio of first class seed pot bacterial classification and fermention medium.
Preferably, the preparation method of described aspergillus oryzae fermented liquid comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 3-8d at temperature is 25-45 DEG C, then in 1-10 DEG C of preservation, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask that 40-150mL liquid submerged culture base is housed, the rotating speed of triangular flask is 60-200 rev/min is that 24-72h cultivated by the shaking table of 20-45 DEG C in temperature, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by inoculum size be 1-20% by liquid shaking bottle strain inoculation in seed tank culture base, be 20-45 DEG C in temperature, mixing speed is 50-200 rev/min, and per minute air flow is under the condition of 0.2-2:1, cultivate 18-72h, obtained first class seed pot bacterial classification;
Described inoculum size is the volume ratio of liquid shaking bottle bacterial classification and liquid submerged culture base; Described air flow is volume and the liquid submerged culture base volume ratio that per minute passes into gas;
D. liquid fermentation and culture: by inoculum size 2-20% by first class seed pot strain inoculation in fermention medium, at temperature 20-45 DEG C, mixing speed 50-200 rev/min, per minute air flow is under 0.2-2:1 condition, cultivates 3-10d, obtained aspergillus oryzae fermented liquid;
Described inoculum size is the volume ratio of first class seed pot bacterial classification and fermention medium;
Described air flow is that per minute passes into the volume of gas and the volume ratio of fermention medium.
The aspergillus oryzae fermented liquid obtained by above method is light yellow to brown, containing manganese peroxidase enzymic activity 100 ~ 10000U/L fermented liquid, endo cellulase enzymic activity 150 ~ 3000U/L, beta-glucanase enzymic activity 150 ~ 3000U/L, 'beta '-mannase enzymic activity 50 ~ 2000U/L.
Manganese peroxidase enzyme testing method: get appropriate aspergillus oryzae fermented liquid, suction filtration, be enzyme liquid; Containing the Sodium.alpha.-hydroxypropionate damping fluid 3.4mL of pH 4.5 50mmol/L in 4mL reaction system, the manganese sulfate solution 0.1mL of 1.6mmol/L, enzyme liquid X mL, when distilled water (0.4-X) mL is preheated to 37 DEG C, adds the H of 1.6mmol/L
2o
2solution 0.1mL starts reaction; At 240nm UV-light place, OD value changing value in assaying reaction 4min; In control group, replace original enzyme liquid to boil deactivation 15min enzyme liquid, replace H with distilled water
2o
2solution, other reactants are constant; Per minute makes the Mn of 1 μm of ol/L
2+be converted into Mn
3+required enzyme amount is 1 enzyme activity unit (U);
Wherein X: enzyme liquid to be measured amasss (≤0.4mL).
Endo cellulase measuring method: add the carboxymethylcellulose sodium solution 1.5mL of 0.5% in test tube, add the enzyme liquid 0.5mL of suitably dilution, in 50 DEG C of water bath heat preservation 30min, add 3 ' 5 one edlefsen's reagent solution 1.5mL after taking-up immediately, boil 5min, take out, distilled water 21.5mL is added after cooling, mixing, surveys the OD value at 520nm place, compares (3 repetitions) with the enzyme liquid boiling deactivation 15min; Carboxymethylcellulose sodium solution and the enzyme liquid of above-mentioned 0.5% is replaced with the Standard glucose solution of 2mL different concns, repeat above-mentioned steps, absorbancy is measured under similarity condition, according to glucose concn and absorbancy drawing standard curve, calculate the sugar obtained by carboxymethyl cellulose according to typical curve; The enzyme amount that 1 cellulase productivity unit definition generates needed for the glucose of 1 μm of ol/L for per minute hydrolyzed carboxymethylcellulo, e is 1 enzyme activity unit (U).
Determining enzymic activity of beta-glucan method: get 2 25ml scale tool plug test tubes, adds 1.5ml 0.7% beta-glucan substrate solution (being prepared by pH value 5.6 acetic acid-sodium acetate buffer solution) respectively, puts in 55 DEG C of water-baths and be incubated 5min; Add 0.5ml in a test tube wherein and suitably dilute enzyme liquid, mixing, is placed in 55 DEG C of water-bath accurate response 30min, adds in 3ml 3 ' 5 one edlefsen's reagent to two test tube, mixing; The enzyme liquid that 0.5mL dilutes on an equal basis is added in another test tube (blank tube), two test tubes are put into 100 DEG C of water-baths simultaneously and react 10min, taking-up is placed in cold water and is cooled to room temperature, add water and be settled to 25mL, capping plug mixes, measure enzyme sample at 520nm place to the absorbancy of blank, calculate enzyme activity; Activity of beta-glucanase unit definition: under condition determination, per minute hydrolysis beta-glucan produces the enzyme amount needed for 1 μm of ol reducing sugar (with glucose meter), is defined as an enzyme activity unit (IU).
Beta-mannase activity determination method: get 2 25mL scale tool plug test tubes, adds 2mL0.5% locust bean gum solution (being prepared by pH 5.0 acetic acid-sodium acetate buffer solution) respectively, puts in 55 DEG C of water-baths and be incubated 5min.Add 0.5mL in a test tube wherein and suitably dilute enzyme liquid, mixing, is placed in 55 DEG C of water-bath accurate response 10min, adds in 2.5mL 3 ' 5 one edlefsen's reagent to two test tube, mixing; In another test tube (blank tube), add enzyme liquid that 0.5mL dilutes on an equal basis two test tubes are put into 100 DEG C of water-baths simultaneously and react 10min, taking-up is placed in cold water and is cooled to room temperature, and add water 5mL, and capping plug mixes, measure enzyme sample at 520nm place to the absorbancy of blank, calculate enzyme activity; Beta-mannase enzyme activity unit defines: 55 DEG C, under pH5.0 condition, the beta-mannase in per minute hydrolysis locust bean gum produces the enzyme amount needed for 1 μm of ol seminose, is defined as an enzyme activity unit (U).
Further improvement, described first class seed pot bacterial classification is after secondary seed tank seed culture, carry out liquid fermentation and culture again, described secondary seed tank seed culture technique is: by inoculum size be 5-20% by first class seed pot strain inoculation in seed tank culture base, be 20-45 DEG C in temperature, mixing speed is 50-200 rev/min, and per minute air flow is under the condition of 0.2-2:1, cultivate 18-72h, obtained secondary seed tank bacterial classification; Described inoculum size is the volume ratio of first class seed pot bacterial classification and liquid submerged culture base; Described air flow is that per minute passes into the volume of gas and the volume ratio of seed tank culture base.
Further improvement, described secondary seed tank bacterial classification is after three grades of seeding tank seed culture, carry out liquid fermentation and culture again, described three grades of seeding tank seed culture techniques are: by inoculum size be 5-20% by secondary seed tank strain inoculation in seed tank culture base, be 20-45 DEG C in temperature, mixing speed is 50-200 rev/min, and per minute air flow is under the condition of 0.2-2:1, cultivate 18-72h, obtained three grades of seeding tank bacterial classifications; Described inoculum size is the volume ratio of secondary seed tank bacterial classification and liquid submerged culture base; Described air flow is that per minute passes into the volume of gas and the volume ratio of seed tank culture base.
The present invention provides a kind of degrading maize straws to prepare the method for maize straw liquid glucose on the other hand, and the method comprises with aspergillus oryzae fermented liquid of the present invention for catalyzer, with H
2o
2solution is that oxygenant is degraded.
Further improvement, described method comprises the steps:
A) corn stalk powder, aspergillus oryzae fermented liquid and water are mixed, preheating;
B) 0.5-5%H is added
2o
2solution, stirs, and degraded obtains mixed solution;
C) by mixed solution suction filtration, collect filtrate, obtain maize straw liquid glucose.
Preferably, degrading maize straws is prepared the concrete grammar of maize straw liquid glucose and is:
1) corn stalk powder, aspergillus oryzae fermented liquid and water are pressed 1:0.2-5:5-50 mixing, 20-60 DEG C of insulation;
2), under mixing speed 30-300 rev/min of condition, stir;
3) in 3-10h, corn stalk powder weight 5-20 0.5-5%H doubly is at the uniform velocity added
2o
2solution is 20-60 DEG C in temperature, and under mixing speed 30-300 rev/min condition, degraded 5-20 hour, obtains mixed solution;
4) by mixed solution suction filtration, collect filtrate, obtain maize straw liquid glucose.
Preferably, the granularity of corn stalk powder is 10-100 order.
Sugar determination method: 3 ' 5-dinitrosalicylic acid system (Zhang Zhiliang, Qu Weijing.Plant physiology experiment instructs, the third edition, Higher Education Publishing House, 129-130 in 2003).
The advantage of this method is adopted to be: with short production cycle, occupied ground is little, production cost is low, sugared yield is high.
The present invention provides the maize straw liquid glucose that a kind of method preparing maize straw liquid glucose by degradation solution degrading maize straws obtains on the other hand, and the sugared yield of this liquid glucose is 20-50g/100g maize straw.
The present invention provides maize straw liquid glucose on the other hand and can be used as the carbon source raw material producing butanols, ethanol or other fermentation industries.
Beneficial effect of the present invention:
The invention provides a kind of aspergillus oryzae fermented liquid, prepare the bacterial classification that uses in fermented liquid by the mechanisms such as united States food and drug administration (FDA), U.S. feed Gong Ding association (AAFCO) assert can the aspergillus oryzae bacterial classification of feed level microbe additive of Direct-fed animal; And containing the necessary enzyme of xylogen in degrading maize straws in this aspergillus oryzae fermented liquid, solve the technical problem that xylogen is not easily degraded; The degradation solution lignin degrading utilizing aspergillus oryzae fermented liquid of the present invention to prepare, utilizes enzyme engineering technology, and add the xylogen in substrate catalytic hydrolysis maize straw and Mierocrystalline cellulose, owing to not having the interference of miscellaneous bacteria, sugared yield is high; The present invention adopts airtight liquid state fermentation, and degraded also can be carried out in retort, controls the pollution of miscellaneous bacteria; And the degradation method cycle of the present invention is short, and place takies little, be applicable to suitability for industrialized production.
Embodiment
Embodiment 1 one kinds of aspergillus oryzae fermented liquids
Containing manganese peroxidase enzymic activity 100U/L, endo cellulase enzymic activity 150U/L, beta-glucanase enzymic activity 150U/L, 'beta '-mannase enzymic activity 50U/L in described aspergillus oryzae fermented liquid; Described aspergillus oryzae fermented liquid take maize straw as main raw material, and aspergillus oryzae is bacterial strain, through the conventionally preparation that test tube enlarged culturing, liquid submerged culture, seeding tank seed culture and liquid fermentation and culture are obtained.
Embodiment 2 one kinds of aspergillus oryzae fermented liquids
Containing manganese peroxidase enzymic activity 1000U/L, endo cellulase enzymic activity 1000U/L, beta-glucanase enzymic activity 500U/L, 'beta '-mannase enzymic activity 200U/L in described aspergillus oryzae fermented liquid; Described aspergillus oryzae fermented liquid prepares by the following method:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 5d at temperature is 30 DEG C, obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by aspergillus oryzae test tube slant spore inoculating in the shaking flask that liquid submerged culture base is housed, cultivates 50h at temperature is 35 DEG C, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by liquid shaking bottle strain inoculation in seed tank culture base, at temperature is 40 DEG C, cultivates 45h, obtained first class seed pot bacterial classification;
D. liquid fermentation and culture: by first class seed pot strain inoculation in fermention medium, at temperature is 25 DEG C, cultivates 7d, obtained aspergillus oryzae fermented liquid.
Embodiment 3 one kinds of aspergillus oryzae fermented liquids
Containing manganese peroxidase enzymic activity 5000U/L, endo cellulase enzymic activity 2000U/L, beta-glucanase enzymic activity 1500U/L, 'beta '-mannase enzymic activity 1000U/L in described aspergillus oryzae fermented liquid; Described aspergillus oryzae fermented liquid prepares by the following method:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 6d at temperature is 25 DEG C, then in 7 DEG C of preservations, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by a ring aspergillus oryzae test tube slant spore inoculating in the 250mL triangular flask that 100mL liquid submerged culture base is housed, the rotating speed of triangular flask is 100 revs/min is that 48h cultivated by the shaking table of 32 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 1g maltose 1.5g peptone 0.3g
Ferrous sulfate 0.004g magnesium sulfate 0.05g Repone K 0.1g;
C. first class seed pot seed culture: by inoculum size be 8% by liquid shaking bottle strain inoculation in seed tank culture base, be 30 DEG C in temperature, mixing speed is 100 revs/min, and per minute air flow is under the condition of 0.5:1, cultivates 48h, obtained first class seed pot bacterial classification; The composition of described seed tank culture base is identical with the composition of liquid submerged culture base;
D. liquid fermentation and culture: by inoculum size 8% by first class seed pot strain inoculation in fermention medium, temperature 25 DEG C, mixing speed 150 revs/min, per minute air flow is under 1:1 condition, cultivates 5d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 5g maltose 1g wood sugar 0.4g
Peptone 1.2g ferrous sulfate 0.01g magnesium sulfate 0.13g
Repone K 0.4 wheat bran 2 Semen Maydis powder 0.3.
Embodiment 4 one kinds of aspergillus oryzae fermented liquids
Containing manganese peroxidase enzymic activity 10000U/L, endo cellulase enzymic activity 3000U/L, beta-glucanase enzymic activity 3000U/L, 'beta '-mannase enzymic activity 2000U/L in described aspergillus oryzae fermented liquid; Described aspergillus oryzae fermented liquid prepares by the following method:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 8d at temperature is 45 DEG C, then in 8 DEG C of preservations, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: liquid submerged culture base is through 125 DEG C of sterilizing 120min, cool to room temperature, by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask of 250mL that 70mL liquid submerged culture base is housed, the rotating speed of triangular flask is 150 revs/min is that 24h cultivated by the shaking table of 20 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 0.5g maltose 0.2g peptone 0.2g
Ferrous sulfate 0.03g magnesium sulfate 0.01g Repone K 0.01g;
C. first class seed pot seed culture: seed tank culture base is through 125 DEG C of sterilizing 120min, cool to room temperature, by inoculum size be 20% by liquid shaking bottle strain inoculation in seed tank culture base, it is 25 DEG C in temperature, mixing speed is 70 revs/min, per minute air flow is under the condition of 2:1, cultivates 72h, obtained first class seed pot bacterial classification; The composition of described seed tank culture base is identical with the composition of liquid submerged culture base;
D. secondary seed tank seed culture: seed tank culture base is through 125 DEG C of sterilizing 120min, cool to room temperature, by inoculum size be 10% by first class seed pot strain inoculation in seed tank culture base, it is 25 DEG C in temperature, mixing speed is 100 revs/min, per minute air flow is under the condition of 0.8:1, cultivates 56h, obtained secondary seed tank bacterial classification;
E. three grades of seeding tank seed culture: seed tank culture base is through 125 DEG C of sterilizing 120min, cool to room temperature, by inoculum size be 10% by secondary seed tank strain inoculation in seed tank culture base, it is 30 DEG C in temperature, mixing speed is 120 revs/min, per minute air flow is under the condition of 1:1, cultivates 48h, obtained three grades of seeding tank bacterial classifications;
F. liquid fermentation and culture: fermention medium through 110 DEG C of sterilizing 50min, cool to room temperature, by inoculum size 4% by three grades of seeding tank strain inoculation in fermention medium, temperature 35 DEG C, mixing speed 200 revs/min, per minute air flow is under 0.2:1 condition, cultivate 3d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 8g maltose 3g wood sugar 3g
Peptone 2g ferrous sulfate 0.02g magnesium sulfate 0.2g
Repone K 1g wheat bran 5g Semen Maydis powder 1g.
The preparation method of embodiment 5 one kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 4d at temperature is 30 DEG C, obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by aspergillus oryzae test tube slant spore inoculating in the shaking flask that liquid submerged culture base is housed, cultivates 29h at temperature is 26 DEG C, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by liquid shaking bottle strain inoculation in seed tank culture base, at temperature is 30 DEG C, cultivates 36h, obtained first class seed pot bacterial classification;
D. liquid fermentation and culture: by first class seed pot strain inoculation in fermention medium, at temperature 30 DEG C, cultivates 7d, obtained aspergillus oryzae fermented liquid;
Obtained aspergillus oryzae fermented liquid is yellow, containing manganese peroxidase enzymic activity 500U/L, and endo cellulase enzymic activity 1500U/L, beta-glucanase enzymic activity 2000U/L, 'beta '-mannase enzymic activity 500U/L.
The preparation method of embodiment 6 one kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 8d at temperature is 25 DEG C, obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by aspergillus oryzae test tube slant spore inoculating in the shaking flask that liquid submerged culture base is housed, cultivates 24h at temperature is 45 DEG C, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 0.2g maltose 1g peptone 0.5g
Ferrous sulfate 0.0005g magnesium sulfate 0.05g Repone K 0.15g;
C. first class seed pot seed culture: by liquid shaking bottle strain inoculation in seed tank culture base, at temperature is 25 DEG C, cultivates 30h, obtained first class seed pot bacterial classification; The composition of seed tank culture base is identical with the composition of liquid submerged culture base;
D. liquid fermentation and culture: by first class seed pot strain inoculation in fermention medium, at temperature 20 DEG C, cultivates 8d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 2g maltose 1g peptone 1g
Ferrous sulfate 0.012g magnesium sulfate 0.09g Repone K 0.3g wheat bran 1g;
Obtained aspergillus oryzae fermented liquid is light yellow, containing manganese peroxidase enzymic activity 7000U/L, and endo cellulase enzymic activity 500U/L, beta-glucanase enzymic activity 1000U/L, 'beta '-mannase enzymic activity 1500U/L.
The preparation method of embodiment 7 one kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 6d at temperature is 30 DEG C, then in 10 DEG C of preservations, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by a ring aspergillus oryzae test tube slant spore inoculating in the 250mL triangular flask that 120mL liquid submerged culture base is housed, the rotating speed of triangular flask is 150 revs/min is that 60h cultivated by the shaking table of 25 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 0.8g maltose 2.5g peptone 0.3g
Ferrous sulfate 0.008g magnesium sulfate 0.01g Repone K 0.13g;
C. first class seed pot seed culture: by inoculum size be 20% by liquid shaking bottle strain inoculation in seed tank culture base, be 30 DEG C in temperature, mixing speed is 50 revs/min, and per minute air flow is under the condition of 0.2:1, cultivates 18h, obtained first class seed pot bacterial classification; The weight of each composition contained by every 100mL seed tank culture base is:
Wheat bran 1.2g maltose 2.2g peptone 0.8g
Ferrous sulfate 0.003g magnesium sulfate 0.12g Repone K 0.17g;
D. liquid fermentation and culture: by inoculum size 5% by first class seed pot strain inoculation in fermention medium, temperature 40 DEG C, mixing speed 70 revs/min, per minute air flow is under 0.2:1 condition, cultivates 6d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 7g maltose 2g wood sugar 2g
Peptone 1.5g ferrous sulfate 0.015g magnesium sulfate 0.1g
Repone K 0.1g wheat bran 4g Semen Maydis powder 0.2g;
Obtained aspergillus oryzae fermented liquid is light brown, containing manganese peroxidase enzymic activity 2000U/L, and endo cellulase enzymic activity 2500U/L, beta-glucanase enzymic activity 2500U/L, 'beta '-mannase enzymic activity 100U/L.
The preparation method of embodiment 8 one kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: a ring aspergillus oryzae bacterial classification is inoculated in potato dextrose medium, cultivates 5d at temperature is 26 DEG C, then in 4 DEG C of preservations, obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: liquid submerged culture base is through 105 DEG C of sterilizing 120min, cool to room temperature, by a ring aspergillus oryzae test tube slant spore inoculating in the 250mL triangular flask that 150mL liquid submerged culture base is housed, the rotating speed of triangular flask is 65 revs/min, be that 26h cultivated by the shaking table of 43 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 2g maltose 0.1g peptone 0.1g
Ferrous sulfate 0.001g magnesium sulfate 0.005g Repone K 0.2g;
C. first class seed pot seed culture: seed tank culture base is through 105 DEG C of sterilizing 120min, cool to room temperature, by inoculum size be 1% by liquid shaking bottle strain inoculation in seed tank culture base, it is 43 DEG C in temperature, mixing speed is 190 revs/min, per minute air flow is under the condition of 0.3:1, cultivates 72h, obtained first class seed pot bacterial classification; The composition of described seed tank culture base is identical with the composition of liquid submerged culture base;
D. liquid fermentation and culture: fermention medium through 130 DEG C of sterilizing 30min, cool to room temperature, by inoculum size 2% by first class seed pot strain inoculation in fermention medium, at temperature 45 C, mixing speed 50 revs/min, per minute air flow is under 2:1 condition, cultivate 3d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 1g maltose 3g wood sugar 0.2g
Peptone 0.3g ferrous sulfate 0.005g magnesium sulfate 0.05g
Repone K 1g wheat bran 0.1g Semen Maydis powder 0.1g;
Obtained aspergillus oryzae fermented liquid is light yellow, containing manganese peroxidase enzymic activity 121U/L, and endo cellulase enzymic activity 3000U/L, beta-glucanase enzymic activity 3000U/L, 'beta '-mannase enzymic activity 2000U/L.
The preparation method of embodiment 9 one kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 3d at temperature is 45 DEG C, then in 1 DEG C of preservation, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask of 250mL that 50mL liquid submerged culture base is housed, the rotating speed of triangular flask is 175 revs/min is that 30h cultivated by the shaking table of 20 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 0.5g maltose 0.5g peptone 0.2g
Ferrous sulfate 0.006g magnesium sulfate 0.009g Repone K 0.05g;
C. first class seed pot seed culture: by inoculum size be 15% by liquid shaking bottle strain inoculation in seed tank culture base, be 45 DEG C in temperature, mixing speed is 200 revs/min, and per minute air flow is under the condition of 2:1, cultivates 35h, obtained first class seed pot bacterial classification; The weight of each composition contained by every 100mL seed tank culture base is:
Wheat bran 2g maltose 0.1g peptone 0.5g
Ferrous sulfate 0.0005g magnesium sulfate 0.12g Repone K 0.2g;
D. secondary seed tank seed culture: by inoculum size be 5% by first class seed pot strain inoculation in seed tank culture base, be 45 DEG C in temperature, mixing speed is 50 revs/min, and per minute air flow is under the condition of 0.2:1, cultivates 18h, obtained secondary seed tank bacterial classification; The weight of each composition contained by every 100mL seed tank culture base is:
Wheat bran 2g maltose 0.1g peptone 0.5g
Ferrous sulfate 0.0005g magnesium sulfate 0.12g Repone K 0.2g;
E. liquid fermentation and culture: by inoculum size 12% by secondary seed tank strain inoculation in fermention medium, temperature 25 DEG C, mixing speed 125 revs/min, per minute air flow is under 0.5:1 condition, cultivates 4d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 3g maltose 2.5g wood sugar 0.05g
Peptone 2.5g ferrous sulfate 0.01g magnesium sulfate 0.12g
Repone K 0.06g wheat bran 0.5g Semen Maydis powder 0.3g;
Obtained aspergillus oryzae fermented liquid is yellow, containing manganese peroxidase enzymic activity 6000U/L, and endo cellulase enzymic activity 700U/L, beta-glucanase enzymic activity 1200U/L, 'beta '-mannase enzymic activity 700U/L.
The preparation method of embodiment 10 1 kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 5d at temperature is 26 DEG C, then in 4 DEG C of preservations, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: liquid submerged culture base is through 120 DEG C of sterilizing 60min, cool to room temperature, by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask of 250mL that 100mL liquid submerged culture base is housed, the rotating speed of triangular flask is 125 revs/min is that 56h cultivated by the shaking table of 34 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 1g maltose 1.5g peptone 0.6g
Ferrous sulfate 0.005g magnesium sulfate 0.1g Repone K 0.1g;
C. first class seed pot seed culture: seed tank culture base is through 120 DEG C of sterilizing 60min, cool to room temperature, by inoculum size be 10% by liquid shaking bottle strain inoculation in seed tank culture base, it is 34 DEG C in temperature, mixing speed is 120 revs/min, per minute air flow is under the condition of 1:1, cultivates 56h, obtained first class seed pot bacterial classification; The composition of described seed tank culture base is identical with the composition of liquid submerged culture base;
D. secondary seed tank seed culture: seed tank culture base is through 120 DEG C of sterilizing 60min, cool to room temperature, by inoculum size be 10% by first class seed pot strain inoculation in seed tank culture base, it is 34 DEG C in temperature, mixing speed is 120 revs/min, per minute air flow is under the condition of 1:1, cultivates 56h, obtained secondary seed tank bacterial classification;
E. liquid fermentation and culture: fermention medium through 120 DEG C of sterilizing 60min, cool to room temperature, by inoculum size 10% by secondary seed tank strain inoculation in fermention medium, temperature 34 DEG C, mixing speed 100 revs/min, per minute air flow is under 1:1 condition, cultivate 7d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 4g maltose 1.5g wood sugar 1.5g
Peptone 2g ferrous sulfate 0.01g magnesium sulfate 0.1g
Repone K 0.5g wheat bran 2.5g Semen Maydis powder 0.5g;
Obtained aspergillus oryzae fermented liquid is light brown, containing manganese peroxidase enzymic activity 4500U/L fermented liquid, and endo cellulase enzymic activity 1400U/L, beta-glucanase enzymic activity 1600U/L, 'beta '-mannase enzymic activity 1100U/L.
The preparation method of embodiment 11 1 kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 4d at temperature is 40 DEG C, then in 7 DEG C of preservations, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask of 250mL that 125mL liquid submerged culture base is housed, the rotating speed of triangular flask is 80 revs/min is that 35h cultivated by the shaking table of 40 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture base is:
Wheat bran 1.7g maltose 3g peptone 0.5g
Ferrous sulfate 0.0009g magnesium sulfate 0.08g Repone K 0.15g;
C. first class seed pot seed culture: by inoculum size be 7% by liquid shaking bottle strain inoculation in seed tank culture base, be 35 DEG C in temperature, mixing speed is 100 revs/min, and per minute air flow is under the condition of 1.5:1, cultivates 60h, obtained first class seed pot bacterial classification; The weight of each composition contained by every 100mL seed tank culture base is:
Wheat bran 0.7g maltose 1g peptone 0.4g
Ferrous sulfate 0.0015g magnesium sulfate 0.09g Repone K 0.1g;
D. secondary seed tank seed culture: by inoculum size be 20% by first class seed pot strain inoculation in seed tank culture base, be 30 DEG C in temperature, mixing speed is 200 revs/min, and per minute air flow is under the condition of 2:1, cultivates 72h, obtained secondary seed tank bacterial classification; The weight of each composition contained by every 100mL seed tank culture base is:
Wheat bran 1.5g maltose 2g peptone 0.1g
Ferrous sulfate 0.001g magnesium sulfate 0.07g Repone K 0.15g;
E. three grades of seeding tank seed culture: by inoculum size be 5% by secondary seed tank strain inoculation in seed tank culture base, be 45 DEG C in temperature, mixing speed is 190 revs/min, and per minute air flow is under the condition of 0.2:1, cultivate 70h, obtained three grades of seeding tank bacterial classifications; The weight of each composition contained by every 100mL seed tank culture base is:
Wheat bran 0.5g maltose 2.5g peptone 0.3g
Ferrous sulfate 0.007g magnesium sulfate 0.05g Repone K 0.05g;
F. liquid fermentation and culture: by inoculum size 18% by three grades of seeding tank strain inoculation in fermention medium, temperature 35 DEG C, mixing speed 80 revs/min, per minute air flow is under 1.5:1 condition, cultivates 9d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 6g maltose 1.7g wood sugar 0.5g
Peptone 1.3g ferrous sulfate 0.008g magnesium sulfate 0.08g
Repone K 0.7g wheat bran 3.5g Semen Maydis powder 0.7g;
Obtained aspergillus oryzae fermented liquid is light brown, containing manganese peroxidase enzymic activity 800U/L fermented liquid, and endo cellulase enzymic activity 2200U/L, beta-glucanase enzymic activity 1300U/L, 'beta '-mannase enzymic activity 1500U/L.
The preparation method of embodiment 12 1 kinds of aspergillus oryzae fermented liquids
Described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 5d at temperature is 26 DEG C, then in 4 DEG C of preservations, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: liquid submerged culture base is through 110 DEG C of sterilizing 70min, cool to room temperature, by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask of 250mL that 40mL liquid submerged culture base is housed, the rotating speed of triangular flask is 200 revs/min is that 72h cultivated by the shaking table of 21 DEG C in temperature, obtained liquid shaking bottle bacterial classification; The weight of each composition contained by every 100mL liquid submerged culture is:
Wheat bran 0.2g maltose 3g peptone 1g
Ferrous sulfate 0.01g magnesium sulfate 0.2g Repone K 0.01g;
C. first class seed pot seed culture: seed tank culture base is through 110 DEG C of sterilizing 70min, cool to room temperature, by inoculum size be 18% by liquid shaking bottle strain inoculation in seed tank culture base, it is 20 DEG C in temperature, mixing speed is 56 revs/min, per minute air flow is under the condition of 1.8:1, cultivates 20h, obtained first class seed pot bacterial classification; The composition of described seed tank culture base is identical with the composition of liquid submerged culture base;
D. secondary seed tank seed culture: seed tank culture base is through 110 DEG C of sterilizing 70min, cool to room temperature, by inoculum size be 18% by first class seed pot strain inoculation in seed tank culture base, it is 20 DEG C in temperature, mixing speed is 56 revs/min, per minute air flow is under the condition of 1.8:1, cultivates 20h, obtained secondary seed tank bacterial classification;
E. three grades of seeding tank seed culture: liquid submerged culture base is through 110 DEG C of sterilizing 70min, cool to room temperature, by inoculum size be 20% by secondary seed tank strain inoculation in seed tank culture base, it is 20 DEG C in temperature, mixing speed is 56 revs/min, per minute air flow is under the condition of 1.8:1, cultivates 20h, obtained three grades of seeding tank bacterial classifications;
F. liquid fermentation and culture: fermention medium is through 105 DEG C of sterilizing 120min, cool to room temperature, by inoculum size 20% by three grades of seeding tank strain inoculation in fermention medium, temperature 22 DEG C, mixing speed 200 revs/min, under per minute air flow 0.3:1 condition, cultivate 10d, obtained aspergillus oryzae fermented liquid;
The weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 8g maltose 0.1g wood sugar 3g
Peptone 3g ferrous sulfate 0.02g magnesium sulfate 0.2g
Repone K 0.01g wheat bran 5g Semen Maydis powder 1g;
Obtained aspergillus oryzae fermented liquid is brown, containing manganese peroxidase enzymic activity 10000U/L fermented liquid, and endo cellulase enzymic activity 150U/L, beta-glucanase enzymic activity 150U/L, 'beta '-mannase enzymic activity 50U/L.
Embodiment 13 degrading maize straws prepares the method for maize straw liquid glucose
Described method comprises the steps:
1) corn stalk powder is broken to 20 orders; Mix by corn stalk powder, aspergillus oryzae fermented liquid and water 1:0.2:5,60 DEG C of insulations;
2), under mixing speed 300 revs/min of conditions, stir;
3) in 10h, the 0.5%H of corn stalk powder weight 20 times is at the uniform velocity added
2o
2solution is 60 DEG C in temperature, under mixing speed 300 revs/min of conditions, degrades 20 hours, obtains mixed solution;
4) by mixed solution suction filtration, collect filtrate, obtain maize straw liquid glucose; Maize straw liquid glucose chromaticness is light yellow, and sugared yield reaches 20g/100g maize straw.
Embodiment 14 degrading maize straws prepares the method for maize straw liquid glucose
Described method comprises the steps:
1) corn stalk powder is broken to 60 orders; Mix by corn stalk powder, aspergillus oryzae fermented liquid and water 1:2.5:25,40 DEG C of insulations;
2), under mixing speed 150 revs/min of conditions, stir;
3) in 6h, the 2.5%H of corn stalk powder weight 12 times is at the uniform velocity added
2o
2solution is 40 DEG C in temperature, under mixing speed 150 revs/min of conditions, degrades 12 hours, obtains mixed solution;
4) by mixed solution suction filtration, collect filtrate, obtain maize straw liquid glucose; Maize straw liquid glucose chromaticness is yellow, and sugared yield reaches 37g/100g maize straw.
Embodiment 15 degrading maize straws prepares the method for maize straw liquid glucose
Described method comprises the steps:
1) corn stalk powder is broken to 100 orders; Mix by corn stalk powder, aspergillus oryzae fermented liquid and water 1:5:50,20 DEG C of insulations;
2), under mixing speed 30 revs/min of conditions, stir;
3) in 3h, the 5%H of corn stalk powder weight 5 times is at the uniform velocity added
2o
2solution is 20 DEG C in temperature, under mixing speed 30 revs/min of conditions, degrades 5 hours, obtains mixed solution;
4) by mixed solution suction filtration, collect filtrate, obtain maize straw liquid glucose; Maize straw liquid glucose chromaticness is brown, and sugared yield reaches 50g/100g maize straw.
The method of embodiment 16 maize straw liquid glucose fermenting alcohol
Maize straw liquid glucose is concentrated into pol and reaches 200g/L; Active Dry Yeast 500g, with 4L 2% glucose solution, 30 DEG C activate 2 hours, and the maize straw liquid glucose that the yeast after activation concentrates with 500L mixes, and 30 DEG C ferment 72 hours; After fermentation ends, 82 DEG C of air distillations, obtain the alcohol of 95%; Spirit yield is 0.14kg alcohol/kg maize straw.
The method of embodiment 17 maize straw liquid glucose fermentation butyl alcohol
Maize straw liquid glucose is concentrated into pol and reaches 80g/L; The strain of butyl alcohol producing of one ring Storage in refrigerator is inoculated into 250mL and 200mL strain of butyl alcohol producing substratum is housed (composition is: maize straw liquid glucose 500mL, peptone 0.2 gram, KH
2pO
40.5 gram, K
2hPO
40.5 gram, FeSO
47H
2o0.01 gram, MgSO
40.02 gram, NaCl 0.01 gram, water 500mL, 120 DEG C of sterilizings 30 minutes), 37 DEG C of quiescent culture 48h, by the inoculum size of 2%, use same substratum further, in seeding tank, air flow 1:1, stir 120 revs/min, temperature 37 DEG C, further enlarged culturing 1-3 time, 1 day time.By 2% inoculum size, seed culture fluid is inoculated in concentrated 121 DEG C of sterilizings stalk hydrolyzed solution of 30 minutes, static 37 DEG C, ferments 6 days, obtain the mixed fermentation liquid containing butanols, ethanol and acetone.Low-temperature atmosphere-pressure distills the butanol mixed liquid obtained containing ethanol, acetone; Mixed solution yield is 0.11kg butanol mixed liquid/1kg Dry corn stalk stalk.
The method of embodiment 18 maize straw liquid glucose fermentation production of protein feedstuff
Maize straw liquid glucose is concentrated into pol and reaches 40g/L; One ring fodder yeast strain inoculation, to liquid shaking bottle seed culture medium (composition is maize straw liquid glucose 1000mL, peptone 5g, wheat bran 1g, 120 DEG C sterilizing 30 minutes) shaking table of 150 revs/min is cultivated 36 hours at 30 DEG C.By the inoculum size of 2%, use same substratum further, in seeding tank, air flow 1:1, stirs 120 revs/min, temperature 37 DEG C, further enlarged culturing 1-4 time, 1 day time; Finally collect thalline, this thalline can as the protein fodder of domestic birds and animals.
Claims (10)
1. an aspergillus oryzae fermented liquid, it is characterized in that, described aspergillus oryzae fermented liquid contains manganese peroxidase enzymic activity 100 ~ 10000U/L, endo cellulase enzymic activity 150 ~ 3000U/L, beta-glucanase enzymic activity 150 ~ 3000U/L, 'beta '-mannase enzymic activity 50 ~ 2000U/L; Described aspergillus oryzae fermented liquid take maize straw as main raw material, and aspergillus oryzae is bacterial strain, obtains through test tube enlarged culturing, liquid submerged culture, seeding tank seed culture and liquid fermentation and culture.
2. a preparation method for aspergillus oryzae fermented liquid according to claim 1, is characterized in that, described method comprises the steps:
A. test tube enlarged culturing: aspergillus oryzae bacterial classification is inoculated in potato dextrose medium, cultivates, obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by aspergillus oryzae test tube slant spore inoculating in the shaking flask that liquid submerged culture base is housed, cultivate, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by liquid shaking bottle strain inoculation in seed tank culture base, cultivate, obtained first class seed pot bacterial classification;
D. liquid fermentation and culture: by first class seed pot strain inoculation in fermention medium, cultivate, obtained aspergillus oryzae fermented liquid.
3. preparation method as claimed in claim 2, it is characterized in that, the weight of every 100mL liquid submerged culture base and each composition contained by every 100mL seed tank culture base is:
Wheat bran 0.2-2g maltose 0.1-3g peptone 0.1-1g
Ferrous sulfate 0.0005-0.01g magnesium sulfate 0.005-0.2g
Repone K 0.01-0.2g;
Preferably, the weight of each composition contained by every 100mL fermention medium is:
Corn stalk powder 1-8g maltose 0.1-3g wood sugar 0-3g
Peptone 0.3-3g ferrous sulfate 0.005-0.02g magnesium sulfate 0.05-0.2g
Repone K 0.01-1g wheat bran 0.1-5 Semen Maydis powder 0-1g;
Preferably, the granularity of described wheat bran is 10-100 order.
4. preparation method as claimed in claim 3, it is characterized in that, described method comprises the steps:
A. test tube enlarged culturing: be inoculated in potato dextrose medium by aspergillus oryzae bacterial classification, cultivates 3-8d at temperature is 25-45 DEG C, then in 1-10 DEG C of preservation, and obtained aspergillus oryzae test tube slant spore;
B. liquid submerged culture: by a ring aspergillus oryzae test tube slant spore inoculating in the triangular flask that 40-150mL liquid submerged culture base is housed, the rotating speed of triangular flask is 60-200 rev/min is that 24-72h cultivated by the shaking table of 20-45 DEG C in temperature, obtained liquid shaking bottle bacterial classification;
C. first class seed pot seed culture: by inoculum size be 1-20% by liquid shaking bottle strain inoculation in seed tank culture base, be 20-45 DEG C in temperature, mixing speed is 50-200 rev/min, and per minute air flow is under the condition of 0.2-2:1, cultivate 18-72h, obtained first class seed pot bacterial classification;
Described inoculum size is the volume ratio of liquid shaking bottle bacterial classification and liquid submerged culture base; Described air flow is volume and the liquid submerged culture base volume ratio that per minute passes into gas;
D. liquid fermentation and culture: by inoculum size 2-20% by first class seed pot strain inoculation in fermention medium, at temperature 20-45 DEG C, mixing speed 50-200 rev/min, per minute air flow is under 0.2-2:1 condition, cultivates 3-10d, obtained aspergillus oryzae fermented liquid;
Described inoculum size is the volume ratio of first class seed pot bacterial classification and fermention medium;
Described air flow is that per minute passes into the volume of gas and the volume ratio of fermention medium.
5. preparation method as claimed in claim 4, it is characterized in that, described first class seed pot bacterial classification is after secondary seed tank seed culture, carry out liquid fermentation and culture again, described secondary seed tank seed culture technique is: by inoculum size be 5-20% by first class seed pot strain inoculation in seed tank culture base, be 20-45 DEG C in temperature, mixing speed is 50-200 rev/min, per minute air flow is under the condition of 0.2-2:1, cultivates 18-72h, obtained secondary seed tank bacterial classification; Described inoculum size is the volume ratio of first class seed pot bacterial classification and liquid submerged culture base, and described air flow is that per minute passes into the volume of gas and the volume ratio of seed tank culture base.
6. preparation method as claimed in claim 5, it is characterized in that, described secondary seed tank bacterial classification is after three grades of seeding tank seed culture, carry out liquid fermentation and culture again, described three grades of seeding tank seed culture techniques are: by inoculum size be 5-20% by secondary seed tank strain inoculation in seed tank culture base, be 20-45 DEG C in temperature, mixing speed is 50-200 rev/min, per minute air flow is under the condition of 0.2-2:1, cultivates 18-72h, obtained three grades of seeding tank bacterial classifications; Described inoculum size is the volume ratio of secondary seed tank bacterial classification and liquid submerged culture base, and described air flow is that per minute passes into the volume of gas and the volume ratio of seed tank culture base.
7. degrading maize straws prepares a method for maize straw liquid glucose, it is characterized in that, described method comprises with aspergillus oryzae fermented liquid according to claim 1 for catalyzer, with H
2o
2solution is that oxygenant is degraded.
8. method as claimed in claim 7: it is characterized in that, described method comprises the steps:
A) corn stalk powder, aspergillus oryzae fermented liquid and water are mixed, preheating;
B) 0.5-5%H is added
2o
2solution, stirs, and degraded obtains mixed solution;
C) by mixed solution suction filtration, collect filtrate, obtain maize straw liquid glucose;
Preferably, described method comprises the steps:
1) corn stalk powder, aspergillus oryzae fermented liquid and water are pressed 1:0.2-5:5-50 mixing, 20-60 DEG C of insulation; Preferably, the granularity of described corn stalk powder is 20-100 order;
2), under mixing speed 30-300 rev/min of condition, stir;
3) in 3-10h, corn stalk powder weight 5-20 0.5-5%H doubly is at the uniform velocity added
2o
2solution is 20-60 DEG C in temperature, and under mixing speed 30-300 rev/min condition, degraded 5-20 hour, obtains mixed solution;
4) by mixed solution suction filtration, collect filtrate, obtain maize straw liquid glucose.
9. the maize straw liquid glucose prepared by method according to claim 8, is characterized in that, the sugared yield of described liquid glucose is 20-50g/100g maize straw.
10. maize straw liquid glucose as claimed in claim 9, is characterized in that, described maize straw liquid glucose can be used as the carbon source raw material producing butanols, ethanol or other fermentation industries.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017133040A1 (en) * | 2016-02-03 | 2017-08-10 | 程雪娇 | Bagasse culture medium and preparation method therefor |
| CN107772049A (en) * | 2016-08-25 | 2018-03-09 | 丹阳西联生物技术有限公司 | A kind of straw feed microbe leaven |
| CN109988717A (en) * | 2019-05-05 | 2019-07-09 | 广西壮族自治区畜牧研究所 | One Aspergillus oryzae bacterial strain and its application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103194506A (en) * | 2012-12-18 | 2013-07-10 | 中山市比克生物科技有限公司 | Method for catalytic production of fructo-oligosaccharides by aspergillus oryzae whole cells |
| CN103444988A (en) * | 2012-05-29 | 2013-12-18 | 江苏大学 | Method for Aspergillus oryzae to degrade straw to produce protein feed |
| CN103451162A (en) * | 2012-05-29 | 2013-12-18 | 江苏大学 | Method for Aspergillus oryzae secretion preparation of manganese peroxidase |
| CN103451161A (en) * | 2012-05-29 | 2013-12-18 | 江苏大学 | Method for Aspergillus oryzae to secrete lignin to prepare peroxidase |
-
2015
- 2015-01-08 CN CN201510008400.1A patent/CN104531640B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103444988A (en) * | 2012-05-29 | 2013-12-18 | 江苏大学 | Method for Aspergillus oryzae to degrade straw to produce protein feed |
| CN103451162A (en) * | 2012-05-29 | 2013-12-18 | 江苏大学 | Method for Aspergillus oryzae secretion preparation of manganese peroxidase |
| CN103451161A (en) * | 2012-05-29 | 2013-12-18 | 江苏大学 | Method for Aspergillus oryzae to secrete lignin to prepare peroxidase |
| CN103194506A (en) * | 2012-12-18 | 2013-07-10 | 中山市比克生物科技有限公司 | Method for catalytic production of fructo-oligosaccharides by aspergillus oryzae whole cells |
Non-Patent Citations (2)
| Title |
|---|
| 李敏 等: "利用米曲霉降解玉米秸秆相应面的优化", 《饲料研究》 * |
| 马光 等: "米曲霉发酵玉米秸秆产纤维素酶饲料条件的优化", 《生物技术》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017133040A1 (en) * | 2016-02-03 | 2017-08-10 | 程雪娇 | Bagasse culture medium and preparation method therefor |
| CN107772049A (en) * | 2016-08-25 | 2018-03-09 | 丹阳西联生物技术有限公司 | A kind of straw feed microbe leaven |
| CN109988717A (en) * | 2019-05-05 | 2019-07-09 | 广西壮族自治区畜牧研究所 | One Aspergillus oryzae bacterial strain and its application |
| CN109988717B (en) * | 2019-05-05 | 2021-09-21 | 广西壮族自治区畜牧研究所 | Aspergillus oryzae strain and application thereof |
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