CN101613692B - Method for preparing immobilized cell carrier from tamarind seed gum and sodium alginate complex gel - Google Patents
Method for preparing immobilized cell carrier from tamarind seed gum and sodium alginate complex gel Download PDFInfo
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- CN101613692B CN101613692B CN2008101263704A CN200810126370A CN101613692B CN 101613692 B CN101613692 B CN 101613692B CN 2008101263704 A CN2008101263704 A CN 2008101263704A CN 200810126370 A CN200810126370 A CN 200810126370A CN 101613692 B CN101613692 B CN 101613692B
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Abstract
The invention relates to the field of polymer materials. The gel raw materials of an immobilized cell carrier which is prepared from bentonite and biological polysaccharide complex gel and disclosed in the patent with the application number of 200710151906.3 comprise carrageenan and guar gum complex natural plant polysaccharide. Compared with the prior art, various properties of the immobilized cell carrier are enhanced, but the curing temperature is higher, and the thermal decomposition temperature as well as the mechanical strength are lower, thereby causing more loss of microbial activities and larger damage rate of a cell carrier ball. In order to further improve the properties, the method adopts the following steps: preparing a water solution with the consistency of 2.4-3.0 percent from tamarind seed gum and sodium alginate complex biological polysaccharide; curing the water solution after cooling in a calcium chloride solution with the consistency of 2.5-3.5 percent and a sodium pyroborate solution with the consistency of 2.5-3.5 percent for the second time; and finally, keeping the cured solution in physiological saline. The prepared immobilized cell carrier has the advantages of high mechanical strength, thermal decomposition temperature and microbial curing rate as well as low damage rate and cost.
Description
Technical field
The present invention relates to the functional high molecule material field, especially the material of fixed cell carrier.
Background technology
Fixed cell carrier is a kind of new type functional macromolecular material that grows up in the last thirty years.The fixed cell carrier ball can make biologically active substance have good processing property and production performance by biologically active substances such as embedding, covalent bonds plant, animal, microorganism cellss, is applied to multiple field.
Chinese scholars is attached great importance to the research of fixed cell carrier.Studied the various fixed carrier, we have applied for " wilkinite, biological polysaccharide plural gel immobilized cell carrier and preparation method thereof " on September 20th, 2007, and application number is 200710151906.3, and this application for a patent for invention has following characteristics:
1. be raw material with composite natral plant polysaccharide such as carrageenin, guar gum and wilkinite, Klorvess Liquid is a solidifying agent, prepares the fixed cell carrier ball.Raw material obtains easily, and cost is lower.
2. carrier ball form homogeneous, diameter is 3～5mm; Physical strength is 1.70～1.90kg/g; X ray confirmation wilkinite is peeled off fully; The DSC test confirms that wilkinite, biological polysaccharide plural gel immobilized cell carrier decomposition temperature are 243.8 ℃; Carrier is more stable under pH2～11 environment.Nontoxic, the good biocompatibility of prepared carrier has good processing property, production performance, resistance toheat and response characteristic.These performances are relatively more outstanding in similar carrier, have been subjected to user's welcome.Also there is the space of continuing raising through putting into practice some performance.1. because the carrageenin self performance determines that solidification value is 35 ℃, and is in the operating process that is cured cell, also bigger to the loss of microorganism active; 2. though the physical strength of solid support material has reached 1.70～1.90kg/g, after repeatedly using repeatedly, also have breakage, cause the loss of microorganism, and then reduced immobilized efficient; 3. through the hot analytical proof of DSC, the decomposition temperature of fixed cell carrier is 243.8 ℃, and resistance toheat is still waiting further raising.
Summary of the invention
The method that the purpose of this invention is to provide a kind of tamarind seed gum and sodium alginate plural gel immobilized cell carrier and preparation thereof.It has advantages such as physical strength height, the pH scope of application are wide, thermostability strong, good biocompatibility this carrier, and the carrier that can be used as immobilized cell is applied to different field such as food, biomedicine, biotechnology, environmental science and Chemical Manufacture.
The preparation method of tamarind seed gum of the present invention and sodium alginate plural gel immobilized cell carrier is as follows:
1) it is 85 ℃～95 ℃ in temperature, the preparation concentration expressed in percentage by weight is 2.4～3.0% the tamarind seed gum and the compound aqueous solution of sodium alginate, tamarind seed gum and sodium alginate mass ratio are (4～5): 1, constant temperature stirs, obtain tamarind seed gum and sodium alginate composite solution, be cooled to room temperature then, standby
2) temperature be 3～5 ℃ while stirring with step 1) in the composite solution of gained to splash into concentration expressed in percentage by weight be that the volume ratio of composite solution and calcium chloride solution is (1～1.5) in 2.5～3.5% the calcium chloride solution; 10, isothermal curing reaction 1～2 hour begins to take shape tamarind seed gum and sodium alginate plural gel immobilized cell carrier,
3) under the room temperature, filter and remove calcium chloride solution, use deionized water wash, inhale then and remove surface-moisture, again the plural gel immobilized cell carrier that begins to take shape is added to concentration expressed in percentage by weight and is in 2.5～3.5% the sodium tetraborate solution, plural gel immobilized cell carrier and sodium tetraborate liquor capacity are than being (1～1.5): 20, solidified 3～4 hours
4) cross elimination sodium tetraborate solution, use deionized water wash, be kept in the physiological saline, promptly make tamarind seed gum and sodium alginate plural gel immobilized cell carrier.
Tamarind seed gum in the technique scheme and sodium alginate all are gelinites, in specific solidifying agent, form the cancellated carrier of porous, if breakage, problem such as softening easily take place in the fixed cell carrier ball made from single sodium alginate, the stability and the physical strength of gel particle are relatively poor, are unfavorable for the repeatedly utilization of immobilized cell; And tamarind seed gum itself separately can't balling-up, does not also use the report of fixing of tamarind seed gum cell carrier at present both at home and abroad.
The present invention has following advantage:
1. the present invention is to be raw material with tamarind seed gum and sodium alginate compound bio polysaccharide and wilkinite, and calcium chloride solution and sodium tetraborate solution are solidifying agent, prepare fixed cell carrier.Starting material are cheap and easy to get, and tamarind seed gum is lower than carrageenin valency, and the preparing carriers method is simple, and solidification value can be reduced to room temperature from 35 ℃, and it is fast at room temperature to operate the gel carrier shaping speed, can reduce the active loss of imbedded microbe.
2. resistance to acids and bases is strong, stable under pH3～11 environment, have outside the advantages such as good processing property, production performance and reactive behavior, the physical strength and the elasticity of solid support material are better, bring up to 2.00-2.20kg/g from 1.70-1.90kg/g, the breakage rate of Shi Yonging reduces greatly repeatedly, has reduced the microorganism loss, has improved immobilization efficiency.The cross-linked structure of macropore is more obvious, and resistance toheat is stronger, and the hot analytical proof carrier of DSC decomposition temperature is 498.2 ℃, has improved 255 ℃ than original application patented technology.As the fixation support of enzyme, microorganism cells, animal and plant cells, application prospect is more wide.
Description of drawings
Fig. 1 is a surperficial SEM photo of the present invention (magnification is 35 times)
Fig. 2 is a surperficial SEM photo of the present invention (magnification is 300 times)
Fig. 3 is an inner SEM photo of the present invention (magnification is 300 times)
Fig. 4 is an inner SEM photo of the present invention (magnification is 1000 times)
Fig. 5 is the DSC thermal analysis curve (decomposition temperature is 498.2 ℃) of solid support material
Embodiment
Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail
Embodiment 1
1) be that 85 ℃, rotating speed are under 500 rev/mins the condition in temperature, the preparation concentration expressed in percentage by weight is 2.4% the tamarind seed gum and the compound aqueous solution of sodium alginate, tamarind seed gum and sodium alginate mass ratio are 5: 1, constant temperature stirred 25 minutes, obtain tamarind seed gum and sodium alginate composite solution, be cooled to room temperature then, standby;
2) under temperature is 3 ℃, 120 rev/mins rotating speed, with the syringe that has No. 7 syringe needles the composite solution of gained in the step 1) being splashed into concentration expressed in percentage by weight is in 2.5% the calcium chloride solution, the volume ratio of composite solution and calcium chloride solution is 1: 10, isothermal curing reaction 2 hours, begin to take shape tamarind seed gum and sodium alginate plural gel immobilized cell carrier
3) under the room temperature, filter and remove calcium chloride solution, with deionized water wash 2 times, inhale with filter paper then and remove surface-moisture, again the plural gel immobilized cell carrier that begins to take shape is added to concentration expressed in percentage by weight and is in 2.5% the sodium tetraborate solution, plural gel immobilized cell carrier that begins to take shape and sodium tetraborate liquor capacity ratio are 1: 20, solidify 3 hours;
4) cross elimination sodium tetraborate solution, use deionized water wash 2 times, be kept in the physiological saline, promptly make tamarind seed gum and sodium alginate plural gel immobilized cell carrier.
Embodiment 2
1) be that 95 ℃, rotating speed are under 600 rev/mins the condition in temperature, the preparation concentration expressed in percentage by weight is 3.0% the tamarind seed gum and the compound aqueous solution of sodium alginate, tamarind seed gum and sodium alginate mass ratio are 4: 1, constant temperature stirred 35 minutes, obtain tamarind seed gum and sodium alginate composite solution, be cooled to room temperature then, standby;
2) under temperature is 5 ℃, 150 rev/mins rotating speed, with the syringe that has No. 7 syringe needles the composite solution of gained in the step 1) being dripped to concentration expressed in percentage by weight is in 3.5% the calcium chloride solution, the volume ratio of composite solution and calcium chloride solution is 1.5: 10, isothermal curing reaction 2 hours begins to take shape tamarind seed gum and sodium alginate plural gel immobilized cell carrier;
3) under the room temperature, filter and remove calcium chloride solution, with deionized water wash 4 times, inhale with filter paper then and remove surface-moisture, again the plural gel immobilized cell carrier that begins to take shape is added to concentration expressed in percentage by weight and is in 3.5% the sodium tetraborate solution, plural gel immobilized cell carrier that begins to take shape and sodium tetraborate liquor capacity ratio are 1.5: 20, solidify 4 hours;
4) cross elimination sodium tetraborate solution, use deionized water wash 4 times, be kept in the physiological saline, promptly make tamarind seed gum and sodium alginate plural gel immobilized cell carrier.
Embodiment 3
1) be that 90 ℃, rotating speed are under 550 rev/mins the condition in temperature, the preparation concentration expressed in percentage by weight is 2.7% the tamarind seed gum and the compound aqueous solution of sodium alginate, tamarind seed gum and sodium alginate mass ratio are 4.5: 1, constant temperature stirred 30 minutes, obtain tamarind seed gum and sodium alginate composite solution, be cooled to room temperature then, standby;
2) under temperature is 4 ℃, 135 rev/mins rotating speed, with the syringe that has No. 7 syringe needles bentonitic composite solution being dripped to concentration expressed in percentage by weight is in 3.0% the calcium chloride solution, the volume ratio of composite solution and calcium chloride solution is 1.25: 10, isothermal curing reaction 1.5 hours begins to take shape tamarind seed gum and sodium alginate plural gel immobilized cell carrier;
3) under the room temperature, filter and remove calcium chloride solution, with deionized water wash 3 times, inhale with filter paper then and remove surface-moisture, again the plural gel immobilized cell carrier that begins to take shape is added to concentration expressed in percentage by weight and is in 3.0% the sodium tetraborate solution, plural gel immobilized cell carrier that begins to take shape and sodium tetraborate liquor capacity ratio are 1.25: 20, solidify 3.5 hours;
4) cross elimination sodium tetraborate solution, use deionized water wash 3 times, be kept in the physiological saline, promptly make wilkinite, tamarind seed gum and sodium alginate plural gel immobilized cell carrier.
Embodiment 1,2,3 prepared products are carried out scanning electron microscopic observation to be confirmed: spheroid inside all is tridimensional network, the surface is porous honeycomb structure, surface irregularity, many folds and projection are arranged, carrier ball form homogeneous, diameter 2～3mm, mechanical test confirms that physical strength is 2.00～2.20kg/g; Hot analytical proof tamarind seed gum of DSC and sodium alginate plural gel immobilized cell carrier decomposition temperature are 498.2 ℃; Carrier is more stable under pH3～11 environment, has all reached goal of the invention.
Claims (1)
1. the preparation method of tamarind seed gum and sodium alginate plural gel immobilized cell carrier, its feature is as follows:
1) it is 85 ℃～95 ℃ in temperature, the preparation concentration expressed in percentage by weight is 2.4～3.0% the tamarind seed gum and the compound aqueous solution of sodium alginate, tamarind seed gum and sodium alginate mass ratio are (4～5): 1, constant temperature stirs, obtain the composite solution of tamarind seed gum and sodium alginate, be cooled to room temperature then, standby
2) temperature be 3～5 ℃ while stirring with step 1) in the composite solution of gained to splash into concentration expressed in percentage by weight be in 2.5～3.5% the calcium chloride solution, the volume ratio of composite solution and calcium chloride solution is (1～1.5): 10, isothermal curing reaction 1～2 hour, begin to take shape tamarind seed gum and sodium alginate plural gel immobilized cell carrier
3) under the room temperature, filter and remove calcium chloride solution, use deionized water wash, inhale then and remove surface-moisture, again the plural gel immobilized cell carrier that begins to take shape is added to concentration expressed in percentage by weight and is in 2.5～3.5% the sodium tetraborate solution, plural gel immobilized cell carrier that begins to take shape and sodium tetraborate liquor capacity are than being (1～1.5): 20, solidified 3～4 hours
4) cross elimination sodium tetraborate solution, use deionized water wash, be kept in the physiological saline, promptly make tamarind seed gum and sodium alginate plural gel immobilized cell carrier.
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| CN101948824B (en) * | 2010-05-24 | 2012-07-18 | 南京林业大学 | Novel technology for granular latex gel complex carrier immobilized cell |
| CN102078637A (en) * | 2011-01-26 | 2011-06-01 | 西北师范大学 | Sodium alginate/tamarind gum blended drug-loaded membrane as well as preparation method and application thereof |
| CN106222160B (en) * | 2016-08-02 | 2019-06-07 | 南京工业大学 | The preparation method and applications of the compound alginate fixation support of molasses colloid |
| CN109207467A (en) * | 2018-11-19 | 2019-01-15 | 南京工业大学 | A kind of method for immobilizing cell of grate sulfur thiobacillus |
| CN113856576B (en) * | 2021-11-02 | 2023-05-30 | 南京工业大学 | Microencapsulated probiotic bacteria prepared from alginate-isolated whey protein-curdlan |
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| CN101157915A (en) * | 2007-09-20 | 2008-04-09 | 西北师范大学 | Benzyl trimethyl organic bentonite, biological polysaccharide composite carrier immobilized brewing yeast cell and preparation method thereof |
| CN101157914A (en) * | 2007-09-20 | 2008-04-09 | 西北师范大学 | Bentonite, biological polysaccharide plural gel immobilized cell carrier and preparation method thereof |
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| CN101157915A (en) * | 2007-09-20 | 2008-04-09 | 西北师范大学 | Benzyl trimethyl organic bentonite, biological polysaccharide composite carrier immobilized brewing yeast cell and preparation method thereof |
| CN101157914A (en) * | 2007-09-20 | 2008-04-09 | 西北师范大学 | Bentonite, biological polysaccharide plural gel immobilized cell carrier and preparation method thereof |
Non-Patent Citations (1)
| Title |
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| 刘慧等.固定化细胞包埋载体－海藻酸盐的应用研究介绍.《齐鲁药事》.2006,第25卷(第9期),544-546. * |
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