CN1888061A - Composite organic-inorganic material film as immobilized cell carrier - Google Patents
Composite organic-inorganic material film as immobilized cell carrier Download PDFInfo
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- CN1888061A CN1888061A CN 200610011073 CN200610011073A CN1888061A CN 1888061 A CN1888061 A CN 1888061A CN 200610011073 CN200610011073 CN 200610011073 CN 200610011073 A CN200610011073 A CN 200610011073A CN 1888061 A CN1888061 A CN 1888061A
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Abstract
The present invention is one kind of composite organic-inorganic material film as immobilized cell carrier. The composite organic-inorganic material film is prepared through adding alumina precursor, such as boehmite, pseudo boehmite, mesoporous molecular sieve MCM-41, etc to polymer material, such as foamed polyurethane, sodium alginate, PVA, etc. for modification. It is superior to traditional immobilized cell carrier material, and has unique pore canal structure for microbe cell to habitat and to propagate. It has coating and adsorbing amount up to 3-6 billion/ml and free fermenting cell up to 0.08-0.16 billion/ml, and may be used for immobilized cell, especially alcohol fermentation to shorten fermentation period and raise alcohol yield. It has the features of high permeability, high mechanical strength, high chemical stability and capacity of being reused, and is one kind of high efficiency immobilized cell carrier.
Description
Technical field
The present invention relates to the cell immobilization carrier material, particularly a kind of composite organic-inorganic material film as immobilized cell carrier that is applied to molasses or the fermentation of starch material distillery yeast.
Background technology
Immobilized cell technology is an emerging biometric technology that grows up the sixties in 20th century, compare with traditional free cell fermentation process, has the processing efficiency height, good stability, the load of microorganisms amount is big, the incomparable advantage of free cell fermentation is arranged, so immobilized cell technology becomes the emphasis of studying over year surplus in the of nearly 10.
Kind of immobilized microorganism activity of such catalysts and fixation support and preparation method thereof has much relations, traditional fixation support material such as polyacrylamide, alginate calcium, polyvinyl chloride, diatomite, pottery etc., have that biochemical stability is poor, mass transfer effect is poor, physical strength is low, easily by biological decomposition and the higher shortcoming of cost, be difficult to meet industrial requirement.United States Patent (USP) 4983299 report is by urethane, absorption activated carbon and as the immobilization biological carrier; Chinese patent CN1105649A is by adding in the polyurethane preparation process behind gac, the foaming as fixed microorganism carrier, Chinese patent CN1478891A adds gac, polyisocyanates, water-retaining agent to improve carrier microorganisms affinity and water-absorbent in the polyether(poly)urethane foaming process, these carriers still exist adsorptive power poor, microorganism affinity, water-absorbent and stability problem still to be strengthened.Development of new fixed microorganism carrier material just requires to overcome the above shortcoming of conventional carriers, carries out modification and modification by preparation novel inorganic-inorganic, composite organic-inorganic material and to material, to regulate carrier loaded performance microorganism is carried out immobilization.Compare with above-mentioned solid support material, urethane, sodium alginate, polyvinyl alcohol etc. have advantages such as nontoxic to microorganism, that the source is abundant, and is cheap, but intensity are relatively poor as embedding carrier commonly used, and specific surface is little.Aluminum oxide and presoma thereof such as pseudo-boehmite, boehmite and mesopore molecular sieve MCM-41, excellent biological compatibility and unique pore passage structure are arranged, can provide microorganism cells suitable growing environment, after modification in the above-mentioned inorganics adding polymer substance, the complex carrier intensity enhancing, has more porous road grid structure, permeability is better, thereby microbial reproduction is speeded up, more in the unit volume, improved zymamsis efficient, and the interpolation modified high-molecular material of these inorganic materials does not appear in the newspapers.
Summary of the invention
The object of the present invention is to provide a kind of unique porous road grid structure, a kind of composite organic-inorganic material film as immobilized cell carrier of formation with unique pore passage structure with the perch of microorganism cells high-density, breeding microenvironment.
Membrane carrier of the present invention is at common polyether(poly)urethane foam, sodium alginate, polyvinyl alcohol (the polymerization degree 1750 ± 50, as follows) etc. add the alumina precursor pseudo-boehmite in the high molecular polymer material, boehmite, inorganic substance such as mesopore molecular sieve, make composite organic-inorganic material, utilize the pore passage structure of these inorganics uniquenesses, and it is good in conjunction with the polymer substance cellular affinity, it is good that the embedding amount reaches inorganic substance intensity greatly, specific surface is big, permeability is good, stability is strong, advantages such as reproducible utilization, coupling with this absorption embedding, obtain having organic and inorganic loose network structure carrier, be beneficial to propagation and mass transfer, wide prospect in industrial application is arranged.
Technical scheme of the present invention is: by add alumina precursor pseudo-boehmite, boehmite and mesopore molecular sieve MCM-41 modification in common polyether(poly)urethane foam or high molecular polymer materials such as sodium alginate and polyvinyl alcohol, the used proportioning raw materials of the compound membrane carrier of this immobilized cell is:
Polyethers 2.50-5.00, silicone oil 0.02-0.05, triethylene diamine 0.01-0.02, stannous octoate 0.01-0.02, tolylene diisocyanate 1.20-2.40, polyvinyl alcohol 3.00-10.00, alumina precursor 0.40-1.00, water 0.08-0.20;
Perhaps sodium alginate 1.00-7.00, polyvinyl alcohol 3.00-10.00, alumina precursor 0.40-1.00, MCM-41 mesopore molecular sieve 0.40-1.00, bacteria suspension 10.00-40.00, water 88-90.
The former production method: for common polyether(poly)urethane foam, according to proportioning raw materials, with polyethers, silicone oil, triethylene diamine, stannous octoate, water after 20-24 ℃ of following mixing and stirring, add polyvinyl alcohol, alumina precursor, MCM-41 mesopore molecular sieve inorganic materials, add tolylene diisocyanate down in stirring, mixing is placed on and is foamed into membranaceously in the mould, promptly makes the microbial fixed carrier finished product; Behind this microbial fixed carrier adding bacteria suspension, put into the molasses culture medium multiplication culture, the instructionization cultivation can be used for alcoholic fermentation process after 3-6 days.
Latter's production method: for sodium alginate, under the condition of 100-120 ℃ of heated and stirred, make it fully dissolve mixing sodium alginate, polyethylene alcohol and water according to proportioning raw materials, interpolation alumina precursor, MCM-41 mesopore molecular sieve inorganic materials mix, after sterilization, adding bacteria suspension stirs, put into the crosslinked 1-5h of calcium chloride solution of 1%-5%wt after filming with film-making machine, put into the molasses culture medium multiplication culture.Above composite organic-inorganic material film as immobilized cell carrier, cultivation propagation through 3-6 days, carrier embedding amount can reach hundred million/mL of 30-50, ooze out free cell reach 1.2 hundred million/more than the mL, can put it in the fermentor tank with molasses is that raw material ferments, and its immobilization fermentation cycle can shorten within the 10h.
The membranaceous carrier of immobilization matrix material provided by the present invention, alumina precursor pseudo-boehmite, boehmite and mesopore molecular sieve MCM-41 modification are added in utilization in high molecular polymer materials such as common polyether(poly)urethane foam, sodium alginate, polyvinyl alcohol, prepare the novel organic-inorganic matrix material.Utilize the pore passage structure of these inorganic substance uniquenesses, the polymer substance cellular affinity is good, it is good that the embedding adsorptive capacity reaches inorganic substance intensity greatly, specific surface is big, permeability is good, stability is strong, advantages such as reproducible utilization, coupling by this embedding absorption, obtain having loose web frame organic and inorganic complex carrier, increased the cell loading amount, and the elasticity and the stability of material, be beneficial to propagation and mass transfer, thereby increased the industrial applications advantage of the membranaceous carrier of immobilization matrix material provided by the invention.
Use it for immobilized cell, particularly be applied to the zymamsis field, it is strong to have a permeability, and physical strength is good, and chemical stability is good, the microorganism excellent affinity, and the embedding adsorptive capacity (reaches the characteristics of 30-60 hundred million/mL) greatly.Its ferment cycle foreshortened in 10 hours, and the carrier alcohol yied can reach 175.50L.Et-OH/M
3.h, the volume alcohol yied can reach 11.00L.Et-OH/M
3.h, can reach more than 1 year reproducible utilization work-ing life.Preparation method of the present invention is simple, is easy to realize that what the present invention was prepared is a kind of fixed cell carrier efficiently.
Description of drawings
Fig. 1 is the moving fixed yeast fermentation of a secondary of the present invention limit afterflow schema
Among the figure, 1-does not have oily main air strainer, and 2-does not have oily secondary air strainer, 3-molasses material-compound tank, 4-storage tank, 5-feed liquid under meter, 6-gas meter, 7-one grade fermemtation jar, 8-second order fermentation, 9-1# liquid level pipe, 10-2# liquid level pipe, 11-fermented liquid storage tank, 12-pump.
Embodiment
Embodiment 1:
Get 0.08 part of 2.50 parts of polyethers, 0.02 part of silicone oil, 0.01 part of triethylene diamine, 0.01 part of stannous octoate, water after 20-24 ℃ of following mixing and stirring according to quality, add 10.00 parts of polyvinyl alcohol, 0.40 part of boehmite, 0.40 part of MCM-41 mesopore molecular sieve, add 1.20 parts of tolylene diisocyanates down in stirring, mixing is placed on and is foamed into the membranaceous microbial fixed carrier that gets in the mould; Adding yeast bacterium this microbial fixed carrier dense be in the bacteria suspension of hundred million/mL of 15-20, puts into the middle multiplication culture of molasses culture medium (added ingredients and YEPD substratum are similar, as follows); After instructionization cultivation 3-6 days, carrier embedding amount can reach hundred million/mL of 30-50, ooze out the free cell number reach 1.0 hundred million/more than the mL, change fermentation stage over to.
At volume is in the column fermentor tank of 1L, and the filling ratio according to 10.00% adds the said fixing carrier, adds the former molasses of 150g/L, and constantly rouses oxygen, carries out batch fermentation, and fermentation period is 5 hours, and remaining total reducing sugar is 7g/L, and alcoholic strength is 9.3% (v/v).Carrier uses and still kept above-mentioned fermentation efficiency in 6 months.Institute's use equipment and process are in the fermentor tank of 300L at two volumes as shown in drawings, and the filling ratio according to 7.50% adds the membranaceous carrier of said fixing matrix material, are the second order fermentation jar with two fermentor tank series connection, with 0.10h
-1The thinning ratio Continuous Flow add the former molasses of 200g/L, residence time 7.5h, karusen alcoholic strength are 11.00% (v/v); Sugar alcohol transformation efficiency 99.87%, carrier alcohol yied are 175.50L.Et-OH/M
3.h, volume alcohol yied 11.00L.Et-OH/M
3.h; Carrier uses and still kept above-mentioned fermentation efficiency in 6 months.
Embodiment 2:
Get 0.18 part of 5.00 parts of polyethers, 0.05 part of silicone oil, 0.02 part of triethylene diamine, 0.02 part of stannous octoate, water after 20-24 ℃ of following mixing and stirring according to quality, add 3.00 parts of polyvinyl alcohol, 1.00 parts of pseudo-boehmites, 1.00 parts of MCM-41 mesopore molecular sieves, add 2.40 parts of tolylene diisocyanates down in stirring, mixing be placed on be foamed in the mould membranaceous.Adding yeast bacterium this microbial fixed carrier dense is in the bacteria suspension of hundred million/mL of 15-20, puts into the molasses culture medium multiplication culture.After instructionization cultivation 3-6 days, carrier embedding amount can reach hundred million/mL of 30-50, ooze out the free cell number reach 1.2 hundred million/more than the mL, change fermentation stage over to.
At volume is in the column fermentor tank of 1L, and the filling ratio according to 15.00% adds fixation support, adds the former molasses of 200g/L, and constantly rouses oxygen, carries out batch fermentation, and fermentation period is 8 hours, and remaining total reducing sugar is 1.96g/L, and alcoholic strength is 12.6% (v/v); Carrier uses and still kept above-mentioned fermentation efficiency in 6 months.Institute's use equipment is identical with embodiment 1.
Embodiment 3:
Get 4.00 parts of sodium alginates according to quality, 8.00 parts of polyvinyl alcohol, 88.00 parts in water down to dissolving and mix, adds 0.40 part of boehmite 100-120 ℃ of stirring heating condition successively, and 0.40 part of MCM-41 mixes; The sterilization postcooling is to room temperature, and adding the yeast bacterium dense is 20 parts of the bacteria suspensions of hundred million/mL of 15-20, after stirring, films thickness 3mm with film applicator; Solidified 4 hours in the calcium chloride solution with film immersion 2%; After the flushing of big water gaging, put into molasses culture medium drum oxygen propagation, cultivate after 40-72 hour, carrier embedding amount can reach hundred million/mL of 30-50, ooze out the free cell number reach 1.6 hundred million/more than the mL, change fermentation stage over to.
At volume is in the column fermentor tank of 1L, and the filling ratio according to 10.00% adds the said fixing carrier, adds the former molasses of 150g/L, and constantly rouses oxygen, carries out batch fermentation, and fermentation period is 5h, and remaining total reducing sugar is 7.80g/L, and alcoholic strength is 9.0% (v/v); Carrier uses and still kept above-mentioned fermentation efficiency in 6 months.Institute's use equipment is identical with embodiment 1.
Embodiment 5:
Get 7.00 parts of sodium alginates according to quality, 3.00 parts of polyvinyl alcohol, 90.00 parts in water down to dissolving and mix, adds 1.00 parts of boehmites in the stirring heating condition, and 1.00 parts of MCM-41 mix; The sterilization postcooling is to room temperature, and adding the yeast bacterium dense is 30 parts of the bacteria suspensions of hundred million/mL of 15-20, after stirring, films thickness 3mm with film applicator; Solidified 3 hours in the calcium chloride solution with film immersion 2%; After the flushing of big water gaging, put into molasses culture medium drum oxygen propagation, cultivate after 40-72 hour, carrier embedding amount can reach hundred million/mL of 30-50, ooze out the free cell number reach 1.2 hundred million/more than the mL, change fermentation stage over to.
At volume is in the column fermentor tank of 1L, and the filling ratio according to 10.00% adds the said fixing carrier, adds the former molasses of 150g/L, and constantly rouses oxygen, carries out batch fermentation, and fermentation period is 5h, and remaining total reducing sugar is 7.80g/L, and alcoholic strength is 9.0% (v/v); Carrier uses and still kept above-mentioned fermentation efficiency in 6 months.Institute's use equipment is identical with embodiment 1.
Claims (1)
1, a kind of composite organic-inorganic material film as immobilized cell carrier, it is characterized in that making: polyethers 2.50-5.00 with following proportion raw material, silicone oil 0.02-0.05, triethylene diamine 0.01-0.02, stannous octoate 0.01-0.02, tolylene diisocyanate 1.20-2.40, polyvinyl alcohol 3.00-10.00, alumina precursor 0.40-1.00, water 0.08-0.20; According to said method the preparation: with polyethers, silicone oil, triethylene diamine, stannous octoate, water after 20-24 ℃ of following mixing and stirring, add polyvinyl alcohol, alumina precursor, MCM-41 mesopore molecular sieve inorganic materials, add tolylene diisocyanate down in stirring, mixing is placed on and is foamed into membranaceously in the mould, promptly makes the microbial fixed carrier finished product;
Perhaps make: sodium alginate 1.00-7.00, polyvinyl alcohol 3.00-10.00, alumina precursor 0.40-1.00, MCM-41 mesopore molecular sieve 0.40-1.00, bacteria suspension 10.00-40.00, water 88-90 with following proportion raw material; Preparation according to said method: under the condition of 100-120 ℃ of heated and stirred, make it fully dissolve mixing sodium alginate, polyethylene alcohol and water, interpolation alumina precursor, MCM-41 mesopore molecular sieve inorganic materials mix, after sterilization, adding bacteria suspension stirs, put into the crosslinked 1-5h of calcium chloride solution of 1%-5%wt after filming with film-making machine, put into the molasses culture medium multiplication culture, cultivation propagation through 3-6 days, make carrier embedding amount reach hundred million/mL of 30-50, ooze out free cell reach 1.2 hundred million/obtain finished product when mL is above.
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| CNB2006100110736A CN100443591C (en) | 2006-07-25 | 2006-07-25 | Composite organic-inorganic material film as immobilized cell carrier |
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|---|---|---|---|
| CNB2006100110736A CN100443591C (en) | 2006-07-25 | 2006-07-25 | Composite organic-inorganic material film as immobilized cell carrier |
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Cited By (11)
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| WO2010111106A1 (en) * | 2009-03-27 | 2010-09-30 | Dow Global Technologies Inc. | Polymeric foam containing alumina boehmite |
| CN101993866A (en) * | 2009-08-20 | 2011-03-30 | 株式会社日立工业设备技术 | Method for producing entrapping immobilization pellets, apparatus for producing the same, and entrapping immobilization pellets |
| CN103951042A (en) * | 2014-04-04 | 2014-07-30 | 北京工业大学 | Preparation and application of ammonia oxidizing bacteria immobilized straight-tubular bioactive filler based on reticulate carrier |
| CN103951050A (en) * | 2014-04-04 | 2014-07-30 | 北京工业大学 | Preparation and application of denitrifying bacteria immobilized straight-tubular bioactive filler based on reticulate carrier |
| CN103951049A (en) * | 2014-04-04 | 2014-07-30 | 北京工业大学 | Anaerobic ammonia oxidizing bacteria fixed-type bioactive fillers, and preparation and application thereof |
| CN103951078A (en) * | 2014-04-04 | 2014-07-30 | 北京工业大学 | Preparation and application of denitrifying bacteria immobilized star-shaped bioactive filler based on reticulate carriers |
| CN104108800A (en) * | 2014-04-04 | 2014-10-22 | 北京工业大学 | Preparation method and application of netted carrier-based sulfur-oxidizing bacteria immobilized fixed bioactive filler |
| CN104108799A (en) * | 2014-04-04 | 2014-10-22 | 北京工业大学 | Preparation method and application of netted carrier-based sulfate reducing bacteria immobilized fixed bioactive filler |
| CN104894097A (en) * | 2015-05-09 | 2015-09-09 | 浙江省农业科学院 | Long-acting water-purifying bacterium capsule and preparation method thereof |
| CN104909463A (en) * | 2015-05-09 | 2015-09-16 | 浙江省农业科学院 | Core-shell structured water purification bacterium capsule and preparation method thereof |
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| US4983299A (en) * | 1989-04-10 | 1991-01-08 | Allied-Signal | Removal of phenols from waste water by a fixed bed reactor |
| JP3284725B2 (en) * | 1993-12-28 | 2002-05-20 | ぺんてる株式会社 | Aluminum oxide thin film and immobilized body of physiologically active substance |
| CN1105649A (en) * | 1994-01-18 | 1995-07-26 | 尹军 | Porous material for water treatment |
| CN100451111C (en) * | 2004-08-27 | 2009-01-14 | 中国科学院沈阳应用生态研究所 | Process for producing fixed particle to repair surface water |
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2006
- 2006-07-25 CN CNB2006100110736A patent/CN100443591C/en not_active Expired - Fee Related
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| RU2527557C2 (en) * | 2009-03-27 | 2014-09-10 | ДАУ ГЛОБАЛ ТЕКНОЛОДЖИЗ ЭлЭлСи | Polymer foam, containing bemite aluminium oxide |
| CN102365319A (en) * | 2009-03-27 | 2012-02-29 | 陶氏环球技术有限责任公司 | Polymeric foam containing alumina boehmite |
| CN102365319B (en) * | 2009-03-27 | 2013-05-08 | 陶氏环球技术有限责任公司 | Polymeric foam containing alumina boehmite |
| WO2010111106A1 (en) * | 2009-03-27 | 2010-09-30 | Dow Global Technologies Inc. | Polymeric foam containing alumina boehmite |
| CN101993866A (en) * | 2009-08-20 | 2011-03-30 | 株式会社日立工业设备技术 | Method for producing entrapping immobilization pellets, apparatus for producing the same, and entrapping immobilization pellets |
| CN104108799A (en) * | 2014-04-04 | 2014-10-22 | 北京工业大学 | Preparation method and application of netted carrier-based sulfate reducing bacteria immobilized fixed bioactive filler |
| CN103951050B (en) * | 2014-04-04 | 2016-03-02 | 北京工业大学 | A kind of denitrifying bacterium immobilization straight-tube shape biological active filling material preparations and applicatio based on mesh carrier |
| CN103951078A (en) * | 2014-04-04 | 2014-07-30 | 北京工业大学 | Preparation and application of denitrifying bacteria immobilized star-shaped bioactive filler based on reticulate carriers |
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| CN104108799B (en) * | 2014-04-04 | 2016-03-02 | 北京工业大学 | A kind of sulfate reduction bacteria immobilization fixed type biological active filler preparations and applicatio based on mesh carrier |
| CN104909463A (en) * | 2015-05-09 | 2015-09-16 | 浙江省农业科学院 | Core-shell structured water purification bacterium capsule and preparation method thereof |
| CN104894097A (en) * | 2015-05-09 | 2015-09-09 | 浙江省农业科学院 | Long-acting water-purifying bacterium capsule and preparation method thereof |
| CN104894097B (en) * | 2015-05-09 | 2018-11-06 | 浙江省农业科学院 | A kind of long-acting water purification bacterium capsule and preparation method thereof |
| CN105461879A (en) * | 2015-11-11 | 2016-04-06 | 浙江省林业科学研究院 | Multipurpose polyurethane effective microorganism solidification carrier |
| CN105461879B (en) * | 2015-11-11 | 2018-01-19 | 浙江省林业科学研究院 | A kind of multipurpose urethane effective microbe solidified carrier |
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