Functional Properties of Mouse Chitotriosidase Expressed in the Periplasmic Space of Escherichia coli.

ABSTRACT: Chitotriosidase (Chit1) is an enzyme associated with various diseases, including Gaucher disease, chronic obstructive pulmonary disease, Alzheimer disease and cystic fibrosis. In this study, we first expressed mouse mature Chit1 fused with V5 and (His)6 tags at the C-terminus (Chit1-V5-His) in the cytoplasm of Escherichia coli and found that most of the expressed protein was insoluble. In contrast, Chit1 tagged with Protein A at the N-terminus and V5-His at the C-terminus, was expressed in the periplasmic space of E. coli as a soluble protein and successfully purified. We evaluated the chitinolytic properties of the recombinant enzyme using 4-nitrophenyl N,N'-diacetyl-β-D-chitobioside [4NP-chitobioside, 4NP-(GlcNAc)2] and found that its activity was comparable to CHO cells-expressed Chit1-

SUBMITTER: Kimura M 

PROVIDER: S-EPMC5055312 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature