Chapter 3 : Methodology

3.1 Materials
- 500g nutrient broth, yeast suspension (mauripan yeast), 5g lysozyme, 500g LB Broth
( Conda Pronadisa), 500g nutrient agar
3.2 Enzyme production
3.2.1 Preparation of glycerol stock
3.2.2 Preparation of agar plate
3.2.3 Inoculum preparation
3.2.4 Grow on production medium (bioreactor)
3.3 Invertase recovery
3.3.1 Centrifugation
3.4 Suspend cell in buffer
3.4.1 Break the cell homogenizer
3.4.2 Effect of lysozyme concentration and incubation period
3.4.3 Analytical methods (invertase assay, protein assay modified lowry method)
3.4.4 Optical density (spectrophotometer)