NATIVE ENZYME MOBILITY SHIFT ASSAY (NEMSA) - A NEW METHOD FOR MONITORING CARBOXYL GROUP MODIFICATION OF CARBOXYMETHYLCELLULASE FROM ASPERGILLUS-NIGER

Authors
RASHID MH SAQIB AAN RAJOKA MI SIDDIQUI KS
Citation
Mh. Rashid et al., NATIVE ENZYME MOBILITY SHIFT ASSAY (NEMSA) - A NEW METHOD FOR MONITORING CARBOXYL GROUP MODIFICATION OF CARBOXYMETHYLCELLULASE FROM ASPERGILLUS-NIGER, Biotechnology techniques, 11(4), 1997, pp. 245-247
Citations number
9
Categorie Soggetti
Biothechnology & Applied Migrobiology
Journal title
Biotechnology techniques → ACNP
ISSN journal
0951208X
Volume
11
Issue
4
Year of publication
1997
Pages
245 - 247
Database
ISI
SICI code
0951-208X(1997)11:4<245:NEMSA(>2.0.ZU;2-0
Abstract
A simple, sensitive, accurate and more informative assay for determini ng the number of modified groups during the course of carboxyl group m odification is described. Monomeric carboxymethylcellulase (CMCase) fr om Aspergillus niger was modified by 1-ethyl-3(3-dimethylaminopropyl)c arbodiimide (EDC) in the presence of glycinamide. The different time-c ourse aliquots were subjected to non-denaturing PAGE and the gel stain ed for CMCase activity. The number of carboxyl groups modified are dir ectly read from the ladder of the enzyme bands developed at given time . This method showed that after 75 min of modification reaction there were five major species of modified CMCases in which 6 to 10 carboxyls were modified.