This Small Business Innovation Research Phase II project will develop recombinant esterases and lipases for use in detergent and other industrial applications. To reduce the need for high temperature wash conditions and thereby conserve energy, the detergent industry has targeted high specific activity/low temperature lipases for use in detergent formulations. During Phase I of this work recombinant `environmental` DNA libraries were constructed from uncultivated bacteria encrusting lipid-rich whale skeletons located in deep sea basins. To subsist on the copious lipids exuding from the whale skeletons in this frigid (2-5 degree C) environment, members of these diverse bacterial communities manufacture cold-adapted enzymes. Recombinant expression clones were discovered from the whale environmental libraries during the Phase I work using high throughput screening methods. In Phase II the esterases and lipases expressed from these clones will be overexpressed and characterized kinetically to determine their suitability for use in detergent and other industrial applications. In addition, the discovery and screening processes will be optimized to generate a large collection of esterase and lipase genes that will serve as templates for a directed evolution project to create enzymes with high activity at low temperatures yet that are stable at high temperatures. This work will result in a collection of stable cold-active enzymes with the long shelf life required for enzymes in detergent formulations. Potential Commercial Applications of the Research Detergent formulations, anti-stalling and ripening agents for food and beverage applications, interesterification processes in chemical industry.
