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  1. 1. Raiyani
  2. 2. <ul><li>Cell immobilization system is an alternative to enzyme immobilization unlike enzyme immobilization where the enzyme is attached to the substrate in immobilized cell system the target cell immobilized. </li></ul>Raiyani
  3. 3. <ul><li>Entrapment </li></ul>Raiyani
  4. 4. <ul><li>Chitosan is linear polysaccharide composed of randamely distributed 1-4 linked D-glucosamide & N-acetyl D-glucosamide. </li></ul><ul><li>Chitosan is a partially deacylated chitin formed by the reaction of chitin with concerted alkali. </li></ul><ul><li>Chitosan is chemically high molecular weight . </li></ul><ul><li>Chitosan cross linked with high molecular weight counter ion in capsules while cross linking with low molecular weight counter ions in globules in which biocatalyst get entrapped. </li></ul>Raiyani
  5. 5. Raiyani
  6. 6. <ul><li>Cell/enzyme suspended in 1%w/v chitosan acetate solution </li></ul><ul><li>gentle mixing </li></ul><ul><li>This suspension added drop wise in to 2%w/v counter ion </li></ul><ul><li>(ph 8.2 maintained adjust with 5 mole alkaly) </li></ul><ul><li>After the beads are collected by detection of supernant or simply filtering the suspension. </li></ul>Raiyani
  7. 7. <ul><li>Biocatalyst mixed with gel forming material. </li></ul><ul><li>Suspension is hydrophobic phase & gelation is induced. </li></ul><ul><li>Now, washing with biocatalyst is added. </li></ul><ul><li>Formation of beads with biocatalyst entrapped are allowed. </li></ul><ul><li>Sediment under the gentle centrifugation in aqueous phase </li></ul><ul><li>The beads are washed with medium until they are free from hydrophobic phase. </li></ul>Raiyani
  8. 8. <ul><li>If necessary formed beads are sieved by metal screens or nylons nets. </li></ul><ul><li>Sieving may required to remove small beads to avoid the operate continous reactor . </li></ul><ul><li>The whole procedure carried out under the sterile condition. </li></ul><ul><li>EXAMPLE: </li></ul><ul><li>Vegetable oil </li></ul><ul><li>paraffin oil </li></ul>Raiyani
  9. 9. <ul><li>Agarose is a linear ,neutral polysaccharide isolated from marine red algae . </li></ul><ul><li>Agarose basically composed of repeating agarobiose units consisting of alternating 1-3 linked D-galactopyranose.&1-4LINKED 3-6 anhydrous L- galactopyranose. </li></ul><ul><li>The polymer shows hysteresis which means it will dissolved in water at high temperature above its gel forming temperature. </li></ul><ul><li>The gel formed is non charged ,porous, resistant towards bacterial degradation & dose not require counter ions for stability. </li></ul>Raiyani
  10. 10. <ul><li>Normally agarose preparation with gelling temperature between 28 & 40c for immobilization of cell. </li></ul><ul><li>Method :agarose solution 2.5%w/w at 40c is mixed with cells. </li></ul><ul><li>The mixture is dispersed in vegetable oil at 40c under magnetic stirring. </li></ul><ul><li>The droplets size can controlled by adjusting RPM of magnetic stirrer. </li></ul><ul><li>Droplet size is 0.5 – 1.0mm are formed. </li></ul><ul><li>The mixture is cooled an ice bath under continuous stirring until the agarose beads are solidified at 15c. </li></ul>Raiyani
  11. 11. <ul><li>Buffer is added & beads are allow to sediment by gentle centrifugation in to aqueous phase. </li></ul><ul><li>Repeated until the beads preparation are free from the organic phase. </li></ul>Raiyani
  12. 12. <ul><li>Carrageenans are naturally occurring hydrocolloid consisting of high molecular weight linear sulfated polysaccharide. </li></ul><ul><li>These prepared by gum extraction from red algae see weeds & widely used in foods & cosmetic industry as gelling thickening & stabilizing agent. </li></ul><ul><li>There are mainly 3 types naturally occurring carrageenans. </li></ul><ul><li>Kappa </li></ul><ul><li>Lota </li></ul><ul><li>lambda </li></ul>Raiyani
  13. 13. <ul><li>Lambda is soluble in cold water. </li></ul><ul><li>It dose not form a thermo gel on hating. </li></ul><ul><li>Kappa& lota carrageens are insoluble in cold water. </li></ul><ul><li>They are form a thermo reversible gel on heating. </li></ul>Raiyani
  14. 14. Raiyani
  15. 15. <ul><li>Kappa carrageenans have been used for immobilization of biocatalyst. </li></ul><ul><li>Large amount of biocatalyst are homogeneously immobilized in carrageenans gel. </li></ul><ul><li>Compound such enzyme protein from leaking out from gel lattice. </li></ul><ul><li>Lower molecular substance are easily pass through gel lattice. </li></ul><ul><li>These reaction is long period. </li></ul>Raiyani
  16. 16. <ul><li>Mixture of kappa carrageenans & cells </li></ul><ul><li>Take a sample& take injector take sample add </li></ul><ul><li>Cool it. With sample container with </li></ul><ul><li>nylon net. </li></ul><ul><li>Separate the layer. Kcl solution& </li></ul><ul><li>add above cooled &remove </li></ul><ul><li>Kcl removed & solution & kcl solution & </li></ul><ul><li>Cube is formed. Beads are Membrane is </li></ul><ul><li>formed. Formed. </li></ul>Raiyani
  17. 17. <ul><li>Take stock solution acrylamide & N N METHYLENE BISACRYLAMIDE in 50mm in hcl buffer.(ph:7) </li></ul><ul><li>This solution mixed with biocatalyst & ammonium sulphate is added. </li></ul><ul><li>This add in Soya oil &completion beads are collected & washed. </li></ul>Raiyani
  18. 18. <ul><li>Enzyme immobilization has received considerable attention for quite sometimes & it’s approach has been extended towards MULTICOMPONENT CELLULAR SYSTEM. </li></ul><ul><li>Porous silica supports were activated for the enzyme such as alkalamine,the carbonyl & phenol derivatives of titanium activated support. </li></ul>Raiyani
  19. 19. <ul><li>Take 1 gram acid washed Punic stone add 3 ml of 15% w/v titanium chloride solution in 15% w/w hydrochloric acid is added. </li></ul><ul><li>The mixture is dried in an over for 48 hours at 45C. </li></ul><ul><li>Obtained oxychloride derivatives is washed with distilled water.& yielding anhydrous oxide derivatives. </li></ul><ul><li>Now , a 2% suspension of yeast cell in 0.02 M Sodium acetate buffer is added . (ph-4.5) </li></ul>Raiyani
  20. 20. <ul><li>Kept aside for 2 hour at 4 celcious. </li></ul><ul><li>The excessive of suspension is then removed &separated. </li></ul><ul><li>Solid washed with distilled water & 10ml of 0.01M sodium acetate buffer. </li></ul><ul><li>The system obtained are considerably stable at 45 degree celcious. </li></ul>Raiyani
  21. 21. <ul><li>BY SSS </li></ul>Raiyani
  22. 22. Raiyani