Biotechnology: Presented By: Shah Chaital J T .y.b.pham Guidance By:chetan Anajwala

BIOTECHNOLOGY Presented by: shah chaital J T .Y.B.PHAM Guidance by:chetan anajwala

BHAGWAN MAHAVIR COLLEGE OF PHARMACY

ENTRAPMENT OF BIOCATALYST IN CHITOSAN 

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Chitosan is a partially deacylated chitin formed by the reaction of chitin with concerted alkali. Chitosan is chemically high molecular weight linear polymer composition of glucosamine 1-4 linkage. 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.

METHOD OF BEADS FORMATION 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.

ENTRAPMENT OF CELLS IN BEADED POLYMERS 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.

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

ENTRAPMENT OF BIOCATALYST IN AGAROSE 

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Agarose is a linear ,neutral polysaccharide isolated from marine red algae. Agarose basically composed of repeating agarobiose units consisting of alternating 1-3 linked Dgalactopyranose.&1-4LINKED 3-6 anhydrous Lgalactopyranose. 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.

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.

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.