Investigating the chitin-binding capability of recombinant ChiC expressed in E. coli

Abstract

The chitinase C protein (ChiC), natively expressed by Pseudomonas aeruginosa PAO1, has been explored as a safe and environmentally-friendly crop protection agent. Previous work has been conducted to clone the chiC gene into pET-28a to create pM3CRYY A3, an inducible expression vector for ChiC with an N-terminal 6xHis-tag for purification. In this study, we expressed ChiC in Escherichia coli BL21 (DE3) transformed with the plasmid and purified the expressed protein using nickel-resin immobilized metal affinity chromatography. We then assessed chitin-binding ability using a chitin resin and showed that recombinant ChiC expressed in E. coli can bind to chitin in vitro. Our findings enable further research on the activity, stability, and applications of recombinant chitinases.