Fusion of cellulose binding domain to the catalytic domain improves the activity and conformational stability of chitinase in Bacilluslicheniformis DSM13.
Chitinase from Bacilluslicheniformis DSM13 consists of an N-terminal catalytic domain (GH) and a C-terminal chitin binding domain (ChBD). A deletion mutant BliGH and a hybrid chitinase BliGH-CeBD were developed using polymerase chain reaction (PCR) to study the role of substrate-binding domain. Both recombinant chitinases retained their ability to bind to glycol-chitin (GC). BliGH was more effective on colloidal chitin (CC) than BliGH-CeBD as evident from the increased V(max) and k(cat) values. The fusion of CeBD improved the affinity to colloidal chitin, activity and conformational stability in BliGH-CeBD when compared with deletion mutant BliGH.
Neeraja
Chilukoti
C
Department of Plant Sciences, University of Hyderabad, Hyderabad-500046, India.
Moerschbacher
Bruno
B
Podile
Appa Rao
AR
D001407:Bacillus / Q000201:enzymology*; D020134:Catalytic Domain; D002482:Cellulose / Q000378:metabolism*; D002688:Chitinases / Q000737:chemistry / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D013379:Substrate Specificity
D002482:Cellulose; D002688:Chitinases