Chitin binding protein (CBP21) in the culture supernatant of Serratia marcescens 2170

A chitin binding protein (CBP21) 21kDa in size, is a major protein in the culture snpernatant when Serratia marcescens 2170 is grown in the presence of chitin, The gene (cbp) for CBP21 was found to be located in a region 1.5 kb downstream of the chiB gene encoding chitinase B. The cbp gene encodes a polypeptide of 197 amino-acids with a calculated size of 21.6 kDa containing a putative signal sequence of 27 amino acids. Comparison of the amino acid sequence of the deduced polypeptide with that of other proteins showed that CBP21 is similar (45.3% amino acid identity) to CHB1 of Streptomyces olivaceoviridis. Purified CBP21 prepared from the periplasmic fraction of Escherichia coli carrying the cloned cbp gene showed its highest binding activity to squid chitin (beta-chitin) followed by colloidal chitin and regenerated chitin. Binding of CBP21 to regenerated chitin was affected by pH, in particular, low pH reduced binding activity markedly. The presence of similar chitin binding proteins in the distantly related microorganisms, Streptomyces and Serratia, suggests a wide distribution of this type of chitin binding protein in chitinolytic microorganisms.