CN1266286C - Enzyme-bacterium complex biocatalyst and preparation method thereof - Google Patents
Enzyme-bacterium complex biocatalyst and preparation method thereof Download PDFInfo
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- CN1266286C CN1266286C CNB031354432A CN03135443A CN1266286C CN 1266286 C CN1266286 C CN 1266286C CN B031354432 A CNB031354432 A CN B031354432A CN 03135443 A CN03135443 A CN 03135443A CN 1266286 C CN1266286 C CN 1266286C
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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Abstract
The present invention relates to a biocatalyst of a composite body of enzymes and fungi, which is a composite body of enzymes and fungi, a high-efficiency biocatalyst using the enzymes as main components and the fungi as auxiliary components, and a compound of an organic solution and an inorganic substance solution. The biocatalyst of a composite body of enzymes and fungi is mainly used for the biologic fermentation processing of organic fertilizers, such as farmyard fertilizers and the like, and can also be used for the fermentation processing of life organic garbage. The biocatalyst of a composite body of enzymes and fungi comprises the main components of an animal enzyme extracting solution, a plant enzyme extracting solution, an inorganic salt solution and an organic substance solution. The present invention also discloses a preparation method of the biocatalyst of a composite body of enzymes and fungi. The enzyme molecular weight number of the biocatalyst of a composite body of enzymes and fungi is from 100, 000 to more than 1, 000, 000, the enzyme activity is from 20, 000u to 80, 000u, and the enzyme activity preservation rate at a room temperature is above 150. The fertilizer effects of excrement organic fertilizers obtained after the biocatalyst of a composite body of enzymes and fungi is processed are increased by 1 to 3 times, and the excrement organic fertilizers are prepared into high-quality organic fertilizers which have no ovum, no bacterium or no odour. The excrement organic fertilizers can be directly used for crops, and can also be used for the production of solid organic fertilizers and organic composite fertilizers.
Description
Technical field: the present invention's (enzyme bacteria complex) is a kind of enzyme bacteria complex, is to be the high-performance bio catalyzer of assisting based on enzyme, bacterium, and the biological fermentation that is mainly used in natural fertilizers such as farm manure is handled, and also can be used for the fermentative processing of domestic organic garbage.
Background technology: China's long-term application inorganic fertilizer, cause the hardening soil, fertility is degenerated, and agricultural product quality descends, environmental pollution, a series of ecological environment problem such as disruption of ecological balance.Therefore fertilizer or compoiste fertilizer are used in increasing consideration in pollution-free food production.The main body fecaluria fertilizer of China's organic manure resource accounts for more than 50% of organic manure resource total amount, and be used to 10% of too late organic fertilizer application total amount that as fertilizer sources uses, trace it to its cause is because the muck fertilizer efficiency is low, stink is heavy, contain a large amount of germs, parasitic ovum, even virus, that is to say unhygienic.In the prior art, many patent documentations of producing fertilizer by the enzymatic conversion organism are arranged, as application number 00122132 " process for preparing dual-acid leaf fertilizer by mixing earthworm with bacteria and enzymolysis ", application number 01136263 " method of producing organic fertilizer by refuse enzymolysis ", application number 99111462 " tangerine bar fertilizer diluted enzyme quick-effective transformation fermentation agent and preparation method thereof ".Application number 9511943 " Znzyme transformed organic fertilizer ", application number 02133485.4 " a kind of proteolytic enzyme bioid agent and preparation method thereof ".
But in these documents, not having a kind of preparation with enzyme is the special-purpose muck of effectively handling, because muck component complexity, enzyme has strict selectivity to the substrate that it acted on, a certain enzyme often can only work to a certain class material, therefore only adding the single enzyme preparation in the fecaluria biological fermentation is not have effect, is exactly poor effect.In existing document, do not find the same or analogous fertile biological enzyme formulation of handling of fecaluria that is exclusively used in as yet with the present invention.
Summary of the invention: first purpose of the present invention is to develop specially a kind of high-performance bio catalyzer enzyme bacteria complex of enzyme bacteria complex at the complicacy of fecaluria component, biochemical treatment to farm manure produces special effect, second purpose provides the technology of preparation enzyme bacteria complex, the 3rd purpose is to allow natural fertilizers such as fecaluria obtain reasonable use, allow environment be purified, lay the foundation for producing cheap fertilizer and compoiste fertilizer.
The present invention is research and the practice of carrying out under the enzyme scientific theory instructs.Complicacy according to the fecaluria component, according to enzyme its substrate that acts on there are optionally characteristics of strictness, the enzyme that determines us to select for use must be a multi-enzyme system, so that not only each component of fecaluria is all had single-minded combination of zymoprotein and substrate (fecaluria), and has cofactor to participate in the catalytic activity of enzyme active center.Simultaneously, because fecaluria component complexity, also always there is pair multienzyme body catalytic activity to play the composition of positive effect in the substrate (fecaluria), can brings into play the tram of catalytic effect with the arrival of the group on the inducible enzyme active centre, thereby play both sides' " tacit agreement " of enzyme and substrate.In order to reduce the cost of preparation, the present invention is conceived to extract crude zyme preparation from animal vegetable tissue, in order to find the multi-enzyme system that can make enzyme-to-substrate (fecaluria) both sides " tacit agreement ", inventor of the present invention makes efficient enzyme bacteria complex biological catalyst through research and the practice in 8 years.As everyone knows, what zymin rose in biochemical reaction is katalysis, at first is that substrate (fecaluria) combines the unsettled intermediate product of generation with enzyme, and intermediate product resolves into high-quality fertilizer product and enzyme then.The catalytic efficiency of enzyme is generally 10 of organic catalyst
6～10
13Doubly, though so the content of enzyme very low, but but still a large amount of substrate (fecaluria) of catalysis react.Because enzyme bacteria complex catalyst aims is strong, enzyme activity is up to 20000～80000u, and enzyme activity reserve force (room temperature) is more than 150, (under certain condition, the enzyme that can transform micromole's substrate in 1 minute is called a unit of enzyme u).
In enzyme bacteria complex catalyzer, except that containing, also be added with reodorant at the required animals and plants zymin of fecaluria fermentation, in the process of fecaluria fermentation, can remove most foul smell.The high temperature that produces in the fecaluria fermenting process can kill worm's ovum, and simultaneously, thereby the N,O-Diacetylmuramidase of multienzyme body can dissolution of bacteria cell walls killing bacteria and virus.Therefore through the fecaluria fertilizer of enzyme bacteria complex catalyzer fermentative processing, the anosis bacterium of odorless does not have the non-harmful high-quality organic fertilizer of worm's ovum basically, has increased substantially the farm manure quality.
Because enzyme is a protein, be to rearrange polypeptide chain by different order with certain space conformation by 20 seed amino acids, each seed amino acid has different side chains, group on its side chain has nothing in common with each other, hydroxyl, carboxyl, sulfydryl etc. are arranged, and these groups play important role aspect enzyme texture image and the catalytic activity keeping.Except the catalysis of amino acid decision enzyme, the enzyme institute metal ion that belongs to conjugated protein, the enzyme catalysis function is played crucial effect equally, therefore in enzyme bacteria complex Preparation of catalysts process, a certain amount of metal ion and ferment mushroom material must be arranged.
We do not separate one by one the various enzymes in the enzyme bacteria complex catalyzer or differentiate, we think if the enzyme bacteria complex biological catalyst of our development can with fertilizer substrate " tacit agreement " such as fecaluria, have specificity and wholesomeness, just reach our purpose.
According to last surface analysis as can be known, the major ingredient in the enzyme bacteria complex catalyzer is enzyme (as the animals and plants zyme extract), bacterium and relevant interpolation composition, as inorganic salts, reodorant, organic solvent and PH conditioning agent etc.In adding composition, consider the nutrition that plant needs as far as possible,, add the acid salt, phosphoric acid of ammonium etc. as far as possible as in inorganic salt, adding in elements such as N.P that plant needs, the PH conditioning agent.Enzyme bacteria complex biological catalyst partly is made up of biocatalysis agent solution and plant extraction liquid two.The biocatalysis agent solution of enzyme bacteria complex is mainly formed (weight percent) by following composition:
1.. animal livers zyme extract 2～6%
2.. animal blood cell zyme extract 1.5～5%
3.. the thick liquid 2.5～6% of allicin
4. inorganic salt solution 20.5～30.5%
5. organic solution 5～8.5%
6. clear water 44.5～54%
Narrate the preparation method of each composition of biocatalysis agent solution of enzyme bacteria complex below:
1. animal livers extracting solution: use the fresh animal liver,, rub into the liver gruel, add water retting as livers such as domestic animal pig oxen and horses sheep, be heated to 90～95 ℃,, constant temperature stirs half an hour, leaves standstill 30 fens kinds, filter, get filtrate, per 500 gram animal liver can obtain the thick zyme extract of 1000ml animal livers.
2. animal blood cell extracting solution: get animal blood such as fresh domestic animal pig oxen and horses sheep, in 3.8% citric acid three sodium solution, stir.The centrifugation slurry of dehematizing is collected red blood corpuscle, use the deionized water lyse red blood cells, and 0～4 ℃ of standing over night, per 1000 restrain fresh animal bloods can obtain the thick zyme extract of 2000ml animal blood.
3. the thick liquid of allicin: be added to 0.1-0.8M CuSO with mashed garlic
4Soaked in the solution 3～5 hours, reflux one hour is filtered, and gets filtrate, and per 100 gram garlics are obtained the thick liquid of 200ml allicin.
4. inorganic salt solution: manganese sulfate, sal epsom, ammonium acid carbonate is soluble in water, add an amount of metal-salt, add the phosphoric acid acidifying at last, inferior manganese 22.5 grams of sulfur acid in every 5000ml inorganic salt solution, sal epsom 207.5 grams, ammonium acid carbonate 700 grams, metal-salt 38.1 grams, biphosphate zinc solution 60 grams of 3% concentration, 12% phosphoric acid, 608 grams.
5. the preparation of organic solution: under the normal temperature, carbonyl diamine, glycerin diglycidyl ether and sodium polyacrylate are dissolved in the clear water, slowly add yeast, ferment then, stir, contain carbonyl diamine 52.5 grams in every 3500ml organic aqueous solution, glycerin diglycidyl ether 11.2 grams, sodium polyacrylate 7 grams.
The front is the prescription of the biocatalysis agent solution part of enzyme bacteria complex, introduces second plant extraction liquid partly below: get the choppings such as branch, leaf, root, stem of plant, mixing adds water, adds Glacial acetic acid and calcium chloride again, soaks a week, and filtrate is got in filtration.
With first partly product---biocatalysis agent solution and second part product---plant extraction liquid cooperate in 1: 2 or 2: 3 ratios, left standstill 3～5 days, promptly get enzyme bacteria complex biological catalyst.
The biocatalysis agent solution optimum formula of enzyme bacteria complex is (weight percent)
1. the animal livers enzyme extract 4.5%
2. animal blood cell enzyme extract 4.5%
3. the thick liquid 2.5% of allicin
4. inorganic salt solution 30%
5. organic solution 7%
6. clear water 51.5%
The enzyme bacteria complex biological catalyst technological process of production such as Fig. 1.
Operating process: referring to Fig. 1. enzyme bacteria complex biological catalyst technological process of production figure.
Machine 1. animal livers zyme extracts respectively; 2. animal blood cell zyme extract, the 3. thick liquid of allicin, 4. inorganic salt solution, 5. organic solution, 6. plant extraction liquid.
With liver enzyme extracting solution 45ml mix with blood cell zyme extract 45ml the 90ml composite enzyme solution, again with the 90ml composite enzyme solution, the 70ml organic solution, the thick liquid of 25ml allicin, the natural loess of 5 grams is added in the 30ml inorganic salt solution, stirs evenly, add clear water to 1000ml, left standstill 6～8 days, and carried out biochemical reaction, produce 1000ml biocatalysis agent solution, the biocatalysis agent solution of again reaction being finished cooperates by 1: 2 or 2: 3 with plant extraction liquid, stir evenly, left standstill 3～5 days, get enzyme bacteria complex solution.From enzyme bacteria complex biocatalysis agent prescription as can be known, enzyme bacteria complex biological catalyst is the enzyme bacteria complex that multi-enzyme system combines with bacterium, is that main bacterium is auxilliary with enzyme, is the compound of organic solution and mineral solution.
Embodiment:
1. animal livers extracting solution: with fresh animal liver (as pork liver) 500 grams, rub into the liver gruel, put into infusion kettle, add the 750ml clear water, stir and add 0.1ml sulfuric acid down, be heated to 90～95 ℃, constant temperature stirred 30 minutes, left standstill 30 minutes, filter, get filtrate, be diluted with water to 1000ml, per 500 gram animal liver can obtain the thick zyme extract of 1000ml animal livers.
2. animal blood cell extracting solution: get fresh animal blood (as pig blood) 1000 grams, add 143ml 3.8% citric acid three sodium solution, stir.The centrifugation slurry of dehematizing, collect red blood corpuscle, wash red blood corpuscle with 0.9% sodium-chlor then, salt solution is removed in centrifugation, collect red blood corpuscle, add the 1000ml deionized water, vigorous stirring 30 minutes, 0～4 ℃ of standing over night, per 1000 gram fresh animal bloods can obtain the thick zyme extract of 1000ml animal blood.
3. the thick liquid of allicin: get among the copper sulfate 200ml that mashed garlic 100 grams are added to 0.1～0.8M concentration and soaked 3～5 hours, in flask, plugged the prolong reflux one hour, cold filtration, remove filter residue, wash precipitation with 0.1M NaCl and dash to the 200ml scale with clear water for several times, per 100 gram garlics are obtained the thick liquid of 200ml allicin.
4. the preparation of organic solution: under the normal temperature,, make solution in 11.2 gram glycerin diglycidyl ethers and the clear water of 7 gram sodium polyacrylates about 3500ml, add 17.5 gram yeast and 5.25 gram ferment, intermittently stir 2～3 times earlier with 52.5 gram carbonyl diamines.Contain carbonyl diamine 52.5 gram glycerin diglycidyl ethers 11.2 grams in every 3500ml solution, sodium polyacrylate 7 grams.
5. the preparation of inorganic salt solution: elder generation packs the 5000ml clear water in the bucket, 22.5 gram manganese sulfates, 207.5 gram sal epsom, 700 gram ammonium acid carbonates are poured in the water, stirring makes dissolving fully, again metal-salt such as green vitriol 38.1 grams are added in the solution, add 60 grams, 3% primary zinc phosphate, the phosphoric acid stirring and dissolving of 608 grams 12% left standstill under the normal temperature 48 hours.Inferior manganese 22.5 grams of every 5000ml inorganic salt solution sulfur acid, sal epsom 207.5 grams, ammonium acid carbonate 700 grams, metal-salt such as green vitriol 38.1 grams, biphosphate zinc solution 60 grams of 3% concentration, phosphoric acid 608 grams of 12% concentration.
6. plant extraction liquid: plant extraction liquid is an example with the radish.Get radish root, stem and leaf 200 grams, chopping adds water 500 grams, adds each 1 gram of Glacial acetic acid and calcium chloride, soaks a week, stirs, and filters, and gets filtrate, and per 200 gram roots of plants cauline leafs make 500 milliliters of plant extraction liquids.
The above-mentioned liver crude extract 45ml for preparing is mixed with blood cell crude extract 45ml, get the 90ml composite enzyme solution, again with the 90ml composite enzyme solution, the 70ml organic solution, the thick liquid of 25ml allicin, the natural loess of 5 grams is added to and adds clear water in the 30ml inorganic salt solution to the 1000ml scale, stir evenly, left standstill 6～8 days, carry out biochemical reaction biocatalysis agent solution, biocatalysis agent solution that reaction is finished and vegetation water mix by 1: 2 or 2: 3 volume ratios, left standstill 3～5 days, product enzyme bacteria complex biological catalyst.
This preparation enzyme bacteria complex biological catalyst enzyme molecular weight reaches hundreds thousand of to more than 1,000,000, is the complex body of plurality of enzymes, and enzyme activity 20000～80000u, enzyme activity storage rate (room temperature) be more than 150, heavy metal content≤0.0005%.
Fecaluria fertilizer after the processing, fertilizer efficiency increases by 1～3 times, becomes the high-quality fertilizer, no worm's ovum, germ, no stink both can be directly used in farm crop, can be used for producing solid organic fertilizer and compoiste fertilizer again.
This preparation combines with fecaluria is fertile, 6～8 days, can finish the enzymatic hydrolysis and fermentation process.
This preparation characteristic is efficient, quick-acting, nontoxic, pollution-free, and cost is low.
Using method;
(1) 100 jin of liquid farm manure adds this biological catalyst of 200～300ml enzyme bacteria complex, stirs, and leaves standstill 6～8 days, again with 1～3 times or 1～5 times of clear water dilution back use.The general usefulness of topdressing.
(2) 100 jin of solid-state or semi-solid farm manures are handled with 300～400ml enzyme bacteria complex biological catalyst, stir, and seal use in about 8 days.With general usage.Generally make base fertilizer.
(3) 10ml enzyme bacteria complex biological catalyst can be converted 13～15 kilograms of clear water, stirs to do the foliage-spray use.
(4) be that core is produced solid organic fertilizer with enzyme bacteria complex biological catalyst, with after can increasing both production and income, improve crop quality, can improve the soil again, reduce cost.Field test proves for many years, and science is used this system product, food crop volume increase about 5～10%, and vegetable crop is increased production more than 20～30%.
Claims (9)
1. biological catalyst that is mainly used in the farm manure biochemical treatment, it is characterized in that enzyme bacteria complex biological catalyst partly is made up of biocatalysis agent solution and plant extraction liquid two, the volume ratio of biocatalysis agent solution and plant extraction liquid is 1: 2 or 2: 3, the prescription of biocatalysis agent solution count by weight percentage into:
1.. animal livers zyme extract 2～6%
2. animal blood cell zyme extract 1.5～5%
3. the thick liquid 2.5～6% of allicin
4. inorganic salt solution 20.5～30.5%
5. organic solution 5～8.5%
6. clear water 44.5～54%.
2. the biological catalyst that is mainly used in the farm manure biochemical treatment according to claim 1 is characterized in that consisting of of biocatalysis agent solution:
1. the animal livers enzyme extract 4.5%
2. animal blood cell enzyme extract 4.5%
3. the thick liquid 2.5% of allicin
4. inorganic salt solution 30%
5. organic solution 7%
6. clear water 51.5%.
3. the making method that is mainly used in the biological catalyst of farm manure biochemical treatment according to claim 1 and 2, it is characterized in that technical process is: produce earlier the animal livers extracting solution respectively, animal blood cell extracting solution, organic solution, inorganic salt solution, the thick liquid of garlic, plant extraction liquid, after with animal livers extracting solution and animal blood cell extracting solution be mixed in proportion composite enzyme solution, again composite enzyme solution and organic solution are added in the prefabricated inorganic salt solution in proportion, biochemical reaction 6～8 days, the reaction finish the biocatalysis agent solution, the biocatalysis agent solution is mixed with plant extraction liquid by 1: 2 or 2: 3, and biochemical reaction 3～5 days must product enzyme bacteria complex biological catalyst.
4. the method for making that is mainly used in the biological catalyst of farm manure biochemical treatment according to claim 3, the method for making that it is characterized in that the thick zyme extract of animal livers are to be twisted into the liver gruel with the fresh animal liver, flood with clear water, under acidic conditions, heat 90～95 ℃, leave standstill, cold filtration is got filtrate.
5. the method for making that is mainly used in the biological catalyst of farm manure biochemical treatment according to claim 3, the method for making that it is characterized in that animal blood cell extracting solution is to use fresh animal blood, the leaching of adding citric acid three sodium solutions, the centrifugation slurry of dehematizing, collect red blood corpuscle, the water lyse red blood cells, 0～4 ℃ of standing over night of low temperature.
6. the method for making that is mainly used in the biological catalyst of farm manure biochemical treatment according to claim 3, the preparation method who it is characterized in that organic solution is dissolved in carbonyl diamine, glycerin diglycidyl ether and sodium polyacrylate in the clear water under the normal temperature, slowly add yeast and ferment, stir.
7. the method for making that is mainly used in the biological catalyst of farm manure biochemical treatment according to claim 3, the preparation method who it is characterized in that the thick liquid of allicin are to get mashed garlic to be added to 0.1-0.8M CuSO
4Soaked in the solution 3～5 hours, reflux one hour, cooling goes filter residue to get filtrate.
8. the method for making that is mainly used in the biological catalyst of farm manure biochemical treatment according to claim 3, the preparation method who it is characterized in that inorganic salt solution is that manganese sulfate, sal epsom, ammonium acid carbonate, green vitriol, primary zinc phosphate, phosphoric acid is soluble in water successively, mix dissolving thoroughly, leave standstill after 48 hours and take.
9. the method for making that is mainly used in the biological catalyst of farm manure biochemical treatment according to claim 3, the method for making that it is characterized in that plant extraction liquid is: get plant rhizome leaf and smash to pieces, use water retting, add Glacial acetic acid and calcium chloride, soak a week, filter, get filtrate, add a small amount of zinc sulfate and manganous sulfate.
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