Secretome project
Identify proteins secreted by Chlorella cultures
Pellet Chlorella (low speed, ~3K x 3 min), resuspend cells in small volume of growth medium. Incubate overnight, pellet cells, characterize proteins in the supernatant
Develop protein concentration methods.
TCA ppt with detergent (DOC, SLS) carrier seems the preferred method to concentrate dilute proteins.
Phenol extraction is an alternate (complementary?) method.
Characterize the proteins by gel electrophoresis.
Stain with Coomassie blue. (IR scan?)
Sensitivity of silver staining may be necessary.
Ultimately characterize proteins by mass spectrometry.
Does supernatant contain beta-glucosaminidase, alpha-glucosidase activities?
Are proteins attached to the outer wall of Chlorella?