Cloning and Expression of Serratia Marcescens Chia in E. Coli & Yeast

Serratia marcescens, a gram negative bacterium, classified in the large family of Enterobacteriaceae, is very efficient in degradation of chitin because of its ability to produce different chitinolytic enzymes which hydrolyze the ¿-1, 4 linkages in the chitin microfibril. In the present study, an attempt was made to isolate the variant chitinase gene from the bacterium and their expression in E. coli and yeast. One hundred and twenty isolates of S. marcescens from the culture collection of the Department of Biotechnology, University of Agricultural Sciences, Dharwad were screened for efficiency on the basis of their chitinolytic activity on colloidal chitin media, enzyme activity by DNSA method and chi profile in comparison with the reference strain Sm141. The variant chiA from S. marcescens AUDS744 was cloned in pTZ57R/T and further sub-cloned into pET32C+ and pYES2/CT.