CN101875928A - Embedding immobilization method for microbial preparation - Google Patents
Embedding immobilization method for microbial preparation Download PDFInfo
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- CN101875928A CN101875928A CN2009100827992A CN200910082799A CN101875928A CN 101875928 A CN101875928 A CN 101875928A CN 2009100827992 A CN2009100827992 A CN 2009100827992A CN 200910082799 A CN200910082799 A CN 200910082799A CN 101875928 A CN101875928 A CN 101875928A
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Abstract
The invention discloses an embedding immobilization method for a microbial preparation. 1 liter of embedding material comprises 70 to 90g of polyvinyl alcohol, 3 to 5g of sodium alginate, 7 to 9g of silicon dioxide and 0.4 to 0.6g of calcium carbonate; 200 to 250ml of microbial biomass is added into each liter of gel solution (each milliliter of the microbial biomass is 7*10<6> to 9*10<6>CFU); 4 to 6g of activated carbon and 4 to 6g of straw powder are added into microbe solution; and the immobilized grain diameter is about 10mm. The embedding immobilization method has the advantages that microbes well grow in immobilized beads; immobilization materials have no toxic or side effects on aquaculture animals; the immobilization method is simple to operate; the immobilization materials are economic and practical and easy to recycle and reuse; immobilized microbes are easy to preserve and transport and if microbes capable of decontaminating wastewater are immobilized by the method, the immobilized microbes have purification effects superior to those in a suspension state.
Description
Technical field
The present invention relates to bioengineering field, relate to a kind of immobilization method of microorganism that is used for particularly.
Background technology
Immobilization technology is since occurring the fifties in last century, through the development of decades. formed comparatively complete theory and method.Immobilization technology has had and its diverse characteristics under unbound state biological catalyst: as with convenient product separation. biological catalyst is recyclable or recycle, and biological catalyst stability improves greatly, and reaction process can obtain strict control etc.Thereby these characteristics reduce the application cost of expensive biological catalyst greatly. it is applied in large-scale commercial production, and wherein the immobilization technology of cell is the extension of enzyme immobilization technology, is one of the important content in modern biological project field.Characteristics such as immobilized cell has that growth is fast, reaction is fast, contamination resistance is strong, can use continuously, convenient product separation more and more are widely used in food, medicine, environmental protection and fermentation industry at present.
The method of immobilized cell mainly is an entrapping method, secondly is crosslinking or method that the two combines.Industrial application is maximum is exactly immobilized cell with the different shape of entrapping method preparation.Entrapping method is that to make cellular invasion enter porous carrier inner or be wrapped in cell in the gel network structure or in the semipermeability film, small molecules substrate and product can freely spread, and cell can not be diffused in the surrounding medium and goes.This method is simple to operate. and the pair cell activity influence is little, be the process for fixation the most widely of research at present, but embedded material can to a certain degree hinder substrate and oxygen diffusion, and inapplicable to macromolecule substrate.
Summary of the invention
The object of the present invention is to provide a kind of embedding immobilization method of microbial preparation.Adopt embedding immobilization method of the present invention little to the microbial growth influence, embedded material can not produce secondary pollution, and economical, simple to operate, and the convenient transportation of the microorganism after the embedding is preserved.
For achieving the above object, process for fixation provided by the invention adopts entrapping method, and its key step is:
A) with after 60-80 ℃ of water dissolution of polyvinyl alcohol usefulness, adding sodium alginate, silicon-dioxide and lime carbonate mix makes gelating soln;
B) in microbial solution, add gac and stalk powder mixing and absorption;
When c) gelating soln for the treatment of step a is cooled to below 35 ℃ and the product that obtains of step b mix, join in the boric acid solution of calcium chloride of pH=6-7, in 8-12 ℃ of following immobilization 10-30 hour, make immobilized spherule, standby after the balling-up with normal saline flushing.
In one embodiment of the invention, the content of polyvinyl alcohol, sodium alginate, silicon-dioxide and lime carbonate is respectively 70-90g/L, 3-5g/L, 7-9g/L, 0.4-0.6g/L.
In one embodiment of the invention, the microbial biomass that adds in every liter of gelating soln is 200-250mL, and every milliliter contains microorganism is 7 * 10
6-9 * 10
6CFU.The microbial inoculant amount is the total amount of all microorganisms.
In one embodiment of the invention, the add-on of gac and stalk powder is respectively 4-6g/L.
In one embodiment of the invention, the particle diameter of immobilized spherule is about 10mm.
In one embodiment of the invention, silicon-dioxide is Powdered, and its fineness is 100～200 orders.
In one embodiment of the invention, stalk powder is cut off for the stalk that will clean, and passes through drying and crushing again, and the fineness after the pulverizing is near the fineness of silicon-dioxide.
In one embodiment of the invention, gac and stalk powder needed the sterilization through 121 ℃, 30 minutes before adding microbial solution.
The present invention adds sodium alginate and not only helps to improve balling property, and makes active the raising, can also prevent adhesion phenomenon, also can play the certain protection effect to microorganism cells, can alleviate the toxic action of boric acid to microorganism; Add silicon-dioxide and can regulate particle proportion and suppress the rising phenomenon that particle is in operation, can gain in strength again and activity; Lime carbonate both can increase permeability, can form Calucium Silicate powder with silicon-dioxide again and further increase the particle operation stability.
Characteristics of the present invention are:
1) though immobilization method of microorganism is a lot of at present, but still there is certain problem, for example a little less than the microorganism supporting capacity after the immobilization, the easy swelling of immobilized spherule and cause secondary pollution, permeability is poor, a series of associated problem such as microorganism is run off easily, death.A certain amount of gac and stalk powder have been added in the immobilization material of this prescription, these two kinds of materials can play the absorption microorganism respectively and the effect of the nutritive substance of microorganism growth is provided, and the supporting capacity of adding immobilized spherule behind two kinds of materials strengthens, and is difficult for advantages such as swelling.
2) activity charcoal powder of Tian Jiaing and stalk powder are cheap, and can play the advantage of recycling, environmental protection to a certain extent.
3) if the microorganism that fixedly has the function of purifying liquid waste with this method, the effect that the finished product after fixing is purified liquid waste is better than suspended state.
Embodiment
Embedded material of the present invention is made up of polyvinyl alcohol, sodium alginate, silicon-dioxide, lime carbonate.Wherein, the content of each material is respectively 70-90g/L, 3-5g/L, 7-9g/L, 0.4-0.6g/L, and adding microbial biomass in every liter of gelating soln is that (every milliliter contains microorganism is 7 * 10 to 200-250mL
6-9 * 10
6CFU), add 4-6g gac and 4-6g stalk powder and in microbial solution.The immobilization particle diameter is about 10mm.
Elaborate for an embodiment below.
Embodiment
A) stalk is rinsed well, is cut off, be placed on again in the blast drier dry, treat stalk dry out fully the back in pulper, blend powdered, then with its sterilization.
B) polyvinyl alcohol that takes by weighing 80g adds dissolving fully under 80 ℃ in the 1000mL distilled water, adds 4g sodium alginate, 8g silicon-dioxide, 0.5g lime carbonate then and mixes.
C) (every milliliter contains microorganism is 8 * 10 at the microbial solution of 250mL
6CFU) added 5g gac, 5g stalk powder mixing and absorption 10 minutes in.
When d) treating that gelating soln is cooled to 35 ℃ above-mentioned two kinds of solution are mixed, be added dropwise in the boric acid solution that contains 2% calcium chloride in (pH=6.7) with glue head dropper then, 10 ℃ of following immobilizations 24 hours, make immobilized spherule, wash 2-3 time with physiological saline after the balling-up.The bead particle diameter is about 10mm,
E) making successful immobilized spherule can preserve in 4 ℃ refrigerator.Before each the use it is shaken in 1% glucose solution after 6 hours and can use for 2-3 time with deionized water rinsing again.
Claims (10)
1. the embedding immobilization method of a microbial preparation, its key step is:
A) with after 60-80 ℃ of water dissolution of polyvinyl alcohol usefulness, adding sodium alginate, silicon-dioxide and lime carbonate mix makes gelating soln;
B) in microbial solution, add gac and stalk powder mixing and absorption;
When c) gelating soln for the treatment of step a is cooled to below 35 ℃ and the product that obtains of step b mix, join in the boric acid solution of calcium chloride of pH=6-7, in 8-12 ℃ of following immobilization 10-30 hour, make immobilized spherule, water flushing after the balling-up.
2. the embedding immobilization method of microbial preparation according to claim 1, wherein, the content of polyvinyl alcohol, sodium alginate, silicon-dioxide and lime carbonate is respectively 70-90g/L, 3-5g/L, 7-9g/L, 0.4-0.6g/L.
3. the embedding immobilization method of microbial preparation according to claim 1, wherein, the microbial biomass that adds in every liter of gelating soln is 200-250mL, every milliliter contains microorganism is 7 * 10
6-9 * 10
6CFU.
4. as the embedding immobilization method of microbial preparation as described in the claim 3, wherein, the microbial inoculant amount is the total amount of all microorganisms.
5. the embedding immobilization method of microbial preparation according to claim 1, wherein, the add-on of gac and stalk powder is respectively 4-6g/L.
6. the embedding immobilization method of microbial preparation according to claim 1, wherein, the particle diameter of immobilized spherule is about 10mm.
7. the embedding immobilization method of microbial preparation according to claim 1, wherein, silicon-dioxide is Powdered, its fineness is 100～200 orders.
8. the embedding immobilization method of microbial preparation according to claim 1, wherein, stalk powder is cut off for the stalk that will clean, and passes through drying and crushing again, and the fineness after the pulverizing is near the fineness of silicon-dioxide.
9. the embedding immobilization method of live microorganism preparation according to claim 1, wherein, gac and stalk powder needed the sterilization through 121 ℃, 30 minutes before adding microbial solution.
10. the embedding immobilization method of live microorganism preparation according to claim 1, wherein, step c uses normal saline flushing.
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| CN2009100827992A CN101875928A (en) | 2009-04-29 | 2009-04-29 | Embedding immobilization method for microbial preparation |
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| CN102583741A (en) * | 2012-03-01 | 2012-07-18 | 南京大学 | Slow-release carbon source material using sodium alginate as base material and preparation method thereof |
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- 2009-04-29 CN CN2009100827992A patent/CN101875928A/en active Pending
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Application publication date: 20101103 |