Heterologous Expression of the Serratia marcescens Chitinolytic Locus in Cupriavidus necator for Chitin-Derived Bioplastic Production
The global accumulation of petroleum-based plastics, coupled with the vast underutilization of chitin-rich shellfish waste, underscores the need for microbial platforms capable of converting renewable waste streams into biopolymers. Cupriavidus necator, a leading producer of polyhydroxyalkanoates (PHAs), cannot naturally degrade chitin, whereas Serratia marcescens possesses a highly efficient chitinolytic system that includes the regulatory-secretory genes chiR, chiB, chiW, and chiX. This project aims to engineer C. necator with this native S. marcescens locus to enable extracellular chitin depolymerization and subsequent assimilation of N-acetyl-D-glucosamine (NAG) for PHA biosynthesis. The cloned construct will be evaluated first in E. coli and subsequently in C. necator to assess functional secretion and chitinase activity using colloidal chitin agar, followed by growth and polymer production studies in liquid media, with PHB quantified through crotonic acid conversion and HPLC analysis. Together, these experiments establish a proof-of-concept system for transforming chitin waste into biodegradable bioplastic precursors.
Library 314
Christopher J Brigham
5089998219
cbrigham@umassd.edu