• Share
  • Email
  • Embed
  • Like
  • Save
  • Private Content
ENTRAPMENT OF CELL IMMOBILIZATION
 

ENTRAPMENT OF CELL IMMOBILIZATION

on

  • 2,624 views

 

Statistics

Views

Total Views
2,624
Views on SlideShare
2,624
Embed Views
0

Actions

Likes
1
Downloads
58
Comments
0

0 Embeds 0

No embeds

Accessibility

Upload Details

Uploaded via as Microsoft PowerPoint

Usage Rights

© All Rights Reserved

Report content

Flagged as inappropriate Flag as inappropriate
Flag as inappropriate

Select your reason for flagging this presentation as inappropriate.

Cancel
  • Full Name Full Name Comment goes here.
    Are you sure you want to
    Your message goes here
    Processing…
Post Comment
Edit your comment

    ENTRAPMENT OF CELL IMMOBILIZATION ENTRAPMENT OF CELL IMMOBILIZATION Presentation Transcript

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