The discovery and characterization of thermophilic esterases have become an important area of research in recent years due to their potential in catalyzing difficult bond formations under exotic solution conditions (i.e., elevated temperature, extreme pH, and inclusion of organic solvent). In this study, an esterase native to <i>Sphaerobacter thermophilus </i>(Sthe_2743) was produced recombinantly as inclusion bodies, refolded and characterized with respect to its pH and temperature optima, ability to catalyze reactions in the presence of a variety of organic solvents, substrate preferences, metal ions, chaotropic agents, and detergents. The enzyme was found to prefer medium chain fatty acyl substrates, slightly alkaline pH, and temperatures above 50 °C. The enzyme was found to be highly stable as it continued to catalyze reactions in the presence of &gt;30% organic solvent and denaturant solutions (i.e., [urea]&gt; 1 M). Michaelis-Menten studies were utilized to determine kinetic constants to better understand how additives effect the enzyme’s ability to catalyze reactions and for comparison to other thermophilic enzymes. We believe that Sthe_2743 will be a useful tool in catalyzing ester bond formation or hydrolysis for the synthesis of fine chemicals or as a laundry detergent additive.

Purification and kinetic analysis of a thermophilic esterase from S<i>phaerobacter thermophilus</i>

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Purification and kinetic analysis of a thermophilic esterase from Sphaerobacter thermophilus