CN1137991C - Process for immobilizing enzyme/microbe with polyvinyl alcohol - Google Patents
Process for immobilizing enzyme/microbe with polyvinyl alcohol Download PDFInfo
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- CN1137991C CN1137991C CNB001321269A CN00132126A CN1137991C CN 1137991 C CN1137991 C CN 1137991C CN B001321269 A CNB001321269 A CN B001321269A CN 00132126 A CN00132126 A CN 00132126A CN 1137991 C CN1137991 C CN 1137991C
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- polyvinyl alcohol
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- mixed
- agent solution
- crosslinking agent
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- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
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Abstract
The present invention belongs to the biological technical field, which comprises: polyvinyl alcohol, sodium alginate, acrylamide and a water phase are mixed, heated in a water bath until melted, and then, the mixture is still placed, and the temperature of the mixture is lowered to the normal temperature to become a polyvinyl alcohol solution; enzyme/microorganism suspension liquid and the polyvinyl alcohol solution are mixed according to certain proportions; a saturated boric acid solution is added into calcium chloride and N, N'-methylene bisacrylamide to prepare a crosslinking agent solution; the mixed solution is dripped into the cooled crosslinking agent solution with an injector or a drip tube so that white spherical granules are formed in the crosslinking agent solution, soaked in the crosslinking agent solution for a plurality of hours and washed with physiological saline. The present invention has the characteristics of simple preparation, high mechanical strength of the granules and long service life of the granules.
Description
Technical field
The invention belongs to biological technical field, particularly a kind of polyvinyl alcohol immobilized enzyme/method of microorganism.
Background technology
Polyvinyl alcohol (polyvinyl alcohol, be called for short PVA) be a kind of novel enzyme/microorganism entrapped immobilized carrier, have series of advantages such as intensity height, good, the antimicrobial decomposability of chemical stability be strong, cheap, be a kind of embedded material, at home and abroad obtained in recent years than extensive studies with practical potentiality.
Polyvinyl alcohol is water-soluble after heating, gelation takes place add additive in its aqueous solution after, or (10 ℃) freezing formation gel at low temperatures, thereby the microorganism embedding is fixed in the gel network.Additive commonly used has boric acid and borax, adds boric acid or borax in polyvinyl alcohol water solution, forms gel by chemical reaction, and its making step is:
(1) with polyvinyl alcohol and water with 1: 5-1: 10 ratios are mixed mutually, and heating in water bath to polyvinyl alcohol dissolves, and leave standstill then and reduce to normal temperature.
(2) with centrifugal back enzyme/microbial suspension in 0.9%NaCl solution, the weight ratio of enzyme/microorganism weight in wet base and NaCl solution is 1: 1-1: 2.This enzyme/microbial suspension is mixed with certain proportion mutually with polyvinyl alcohol solution.
(3) with syringe mixing solutions is added drop-wise in the saturated boric acid solution of refrigerative, forms the white spherical granules of several millimeters of diameters.
(4) particle is soaked for some time in saturated boric acid solution, wash with physiological saline then.
But the subject matter that exists in this polyvinyl alcohol immobilized method is at present:
One, polyvinyl alcohol are a kind of high viscosity substance, and polyvinyl alcohol and acid reaction are slower, two drops that are more or less the same drop time can bond together when colliding, and progressively be fused into one, promptly granule of polyvinyl alcohol has very strong agglomeration tendency, makes polyvinyl alcohol gel balling-up difficulty;
They are two years old, when polyvinyl alcohol and boric acid react fixedly, three hydroxyls on the boric acid only partly react, also residual on the superpolymer gel that reaction back forms have a hydrophilic hydroxyl, make immobilization particle in application, have very big water-soluble swelling property, along with the growth of duration of service, intensity weakens greatly, and is totally unfavorable in actual applications.
Summary of the invention
The objective of the invention is to solve in the immobilization particle preparation process that exists in the existing polyvinyl alcohol immobilized technology problem that causes the physical strength reduction in be shaped difficulty and the use owing to water-soluble swelling property, a kind of polyvinyl alcohol immobilized enzyme/method of microorganism of practicality is proposed, make it have preparation simply, particulate physical strength height, the characteristics of long service life.
A kind of polyvinyl alcohol immobilized enzyme/method of microorganism that the present invention proposes may further comprise the steps:
(1) polyvinyl alcohol, sodium alginate, acrylamide is mixed with water, heating in water bath is to dissolving, and leaves standstill then to reduce to normal temperature and become polyvinyl alcohol solution; Wherein, the polyvinyl alcohol add-on is 10-20%; The sodium alginate add-on is 0.1-2.0%; The acrylamide add-on is 0.5-9.0%; Its surplus is a water;
(2) enzyme/microbial suspension is mixed with certain proportion mutually with said polyvinyl alcohol solution;
(3) with the N of saturated boric acid solution adding 1-2% calcium chloride and 0.1-1.0%, N '-methylene-bisacrylamide prepares cross-linking agent solution;
(4) mixing solutions that (3) step was prepared with syringe or dropper is added drop-wise in the refrigerative cross-linking agent solution, makes it form white spherical granules, and soaks some hrs therein, washs with physiological saline then.
Characteristics of the present invention and effect:
(1) by in polyvinyl alcohol-boric acid cross-linking process, introducing the agglomeration problem that a spot of (0.1-1.0%) sodium alginate has successfully solved granule of polyvinyl alcohol, strengthened the balling-up ability of polyvinyl alcohol gel;
(2) by in polyvinyl alcohol-boric acid polymerization process, introducing the acrylamide polymerization reaction, make it to form the polyacrylamide reticulated structure in polyvinyl alcohol gel granule interior and surface, improved the water-soluble swelling property of polyvinyl alcohol gel effectively.Polymerization single polymerization monomer acrylamide and linking agent N, the add-on of N '-methylene-bisacrylamide (BIS) is respectively 0.5-9.0% and 0.1-1.0%.
(3) polyvinyl alcohol immobilized technology of the present invention can make the immobilization particle preparation simple, and the particulate physical strength obviously improves, and prolong work-ing life.
Embodiment
Embodiment 1
Preparation process:
(1) 10g polyvinyl alcohol, 0.5g sodium alginate, 6.0g acrylamide are mixed with the 100mL water, heating in water bath leaves standstill then and reduces to normal temperature to dissolving;
(2) cellulase weight in wet base 5g is suspended among the 10mL 0.9%NaCl, mixes mutually with the above-mentioned solution of 12.5mL.
(3) saturated boric acid solution is added 1%CaCl
2With 0.3% N, N '-methylene-bisacrylamide prepares the Chengdu cross-linking agent solution;
(4) syringe with 0.40mm ID is added drop-wise to the mixed solution in (2) in the refrigerative cross-linking agent solution, makes its white ball-type globule that forms the 2mm diameter, and soaks 20h therein, washs with physiological saline then.
Embodiment 2
Preparation process:
(1) 15g polyvinyl alcohol, 1.0g sodium alginate, 4.0g acrylamide are mixed with the 100mL water, heating in water bath leaves standstill then and reduces to normal temperature to dissolving;
(2) trichosporon wet cell weight 5g is suspended among the 12mL 0.9%NaCl, mixes mutually with the above-mentioned solution of 10mL.
(3) saturated boric acid solution is added 2%CaCl
2With 0.6% N, N '-methylene-bisacrylamide prepares the Chengdu cross-linking agent solution;
(4) with the rubber suction pipe mixed solution in (2) is added drop-wise in the refrigerative cross-linking agent solution, makes its white ball-type globule that forms the 3mm diameter, and soak 24h therein, wash with physiological saline then.
Claims (1)
1, a kind of polyvinyl alcohol immobilized enzyme/method of microorganism may further comprise the steps:
(1) polyvinyl alcohol, sodium alginate, acrylamide is mixed with water, heating in water bath is to dissolving, and leaves standstill then to reduce to normal temperature and become polyvinyl alcohol solution; Wherein, the polyvinyl alcohol add-on is 10-20%; The sodium alginate add-on is 0.1-2.0%; The acrylamide add-on is 0.5-9.0%; Its surplus is a water;
(2) enzyme/microbial suspension is mixed with certain proportion mutually with said polyvinyl alcohol solution;
(3) with the N of saturated boric acid solution adding 1-2% calcium chloride and 0.1-1.0%, N '-methylene-bisacrylamide prepares cross-linking agent solution;
(4) mixing solutions that (3) step was prepared with syringe or dropper is added drop-wise in the refrigerative cross-linking agent solution, makes it form white spherical granules, and soaks some hrs therein, washs with physiological saline then.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001321269A CN1137991C (en) | 2000-12-15 | 2000-12-15 | Process for immobilizing enzyme/microbe with polyvinyl alcohol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB001321269A CN1137991C (en) | 2000-12-15 | 2000-12-15 | Process for immobilizing enzyme/microbe with polyvinyl alcohol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1302863A CN1302863A (en) | 2001-07-11 |
| CN1137991C true CN1137991C (en) | 2004-02-11 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB001321269A Expired - Fee Related CN1137991C (en) | 2000-12-15 | 2000-12-15 | Process for immobilizing enzyme/microbe with polyvinyl alcohol |
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| Country | Link |
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| CN (1) | CN1137991C (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100340663C (en) * | 2004-08-06 | 2007-10-03 | 天津大学 | Nano superfine fiber membrane of polyvinyl alcohol with immovable cellulase |
| CN101497880B (en) * | 2009-03-11 | 2011-11-09 | 清华大学 | Novel method for improving PVA immobilized microorganism |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100359010C (en) * | 2004-12-22 | 2008-01-02 | 中国科学院沈阳应用生态研究所 | Method for preparing microbe carrier for immobilization of Flavobacterium through refrigerating-defrosting twice |
| CN1920018B (en) * | 2005-08-24 | 2012-01-11 | 兰州大学 | Preparation method of polyvinyl alcohol immobilized cell spheroid carrier |
| CN102533713B (en) * | 2011-12-27 | 2013-10-02 | 浙江温州轻工研究院 | Preparation method of ceramic-based biological component having biological activity |
| CN103450496B (en) * | 2012-05-31 | 2015-05-20 | 清华大学 | Immobilized laccase microballoon carrier and its preparation method and use |
| CN102816333B (en) * | 2012-09-05 | 2014-02-19 | 安徽皖维集团有限责任公司 | Preparation method of high-concentration PVA (polyvinyl alcohol) solution |
| CN105994448A (en) * | 2016-06-23 | 2016-10-12 | 袁春华 | Preparation method of efficient insect-proofing and antibacterial seed coating |
| CN109593627B (en) * | 2019-02-21 | 2021-10-22 | 卡登堡酒业(中国)有限公司 | Electric field strengthening brewing process of sea-buckthorn and Chinese wolfberry health-care fruit wine |
-
2000
- 2000-12-15 CN CNB001321269A patent/CN1137991C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100340663C (en) * | 2004-08-06 | 2007-10-03 | 天津大学 | Nano superfine fiber membrane of polyvinyl alcohol with immovable cellulase |
| CN101497880B (en) * | 2009-03-11 | 2011-11-09 | 清华大学 | Novel method for improving PVA immobilized microorganism |
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| Publication number | Publication date |
|---|---|
| CN1302863A (en) | 2001-07-11 |
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