Effect of temperature (A), water content (B), enzyme loading (C), and methanol molar ratio (D) on the LipIAF5-2 WCB activity.

<p>A) Effect of temperature on the hydrolytic activity of the LipIAF5-2 WCB. Enzyme activities are expressed relative to maximal activity observed at 40°C (full squares). B) Effect of water content by percentage of substrate weight. Enzyme activities are expressed as final conversion of 1 mmol of glyceryl tributyrate and 4 mmol of methanol after 6 hours of incubation at 40°C. C) Effect of enzyme loading on the conversion of 1 mmol of glyceryl tributyrate using 4 molar equivalents of methanol as acyl acceptor in the aforementionned conditions. Conversion of glyceryl tributyrate (open circles) is expressed as final percentage of theoretical maximal yield (3 mmoles of product) after 3 hours of reaction. Initial specific activity (full squares) is expressed as the slope of liberated product determined under linear conditions over a period of at least 30 minutes. Biocatalyst amounts correspond to 1.65, 3.30, 6.61, 9.92 and 16.53% of substrate weight, respectively. D) Effect of alcohol molar ratios on the LipIAF5-2 WCB initial activity. Initial activity was determined in linear conditions over a 3-hour period using 10 mg of WCB, 5% water (w/w), 1 mmol of glyceryl tributyrate and various molar ratios (alcohol/substrate) of methanol in a final volume of 1.5 mL.