Question

Which chemical is used for extracting enzymes by osmotic shock treatment?

(a) EDTA

(b) NaoH

(c) SDS

(d) 1.4% buffered sucrose

I got this question in semester exam.

This key question is from Extraction of Enzymes by Chemical Methods topic in portion Enzyme Extraction and Isolation of Enzyme Technology

Answer 1

Correct option is (d) 1.4% buffered sucrose

To elaborate: 1.4% buffered sucrose is used to extract enzyme by osmotic method. Washing the cells and equilibrating with this solution leads to bursting and releasing the content outside. Centrifugation is then performed to remove cell debris and extract enzymes separately. EDTA acts as chelating agent which chelate ions present in the cell wall. NaOH is an alkali and acts by nucleophilic attack. SDS is an anionic detergent that solubilizes the phospholipids and lipoproteins present in the cell wall.