Application of design-of-experiments procedures to optimize efficiently pretreatment of lipase for use in a nonaqueous reaction.
A variety of different pretreatments can improve the performance of enzymes in nonpolar reaction media. These pretreatments have primarily been studied in isolation; however, interactions between some pairs of pretreatments are known to exist. The presence of these interactions complicates the design of an optimum multifactor pretreatment. Modern design-of-experiments techniques allow the simultaneous optimization of two or more variables. To improve the performance of lipase in a model reaction, we used a technique called the method of steepest ascent to optimize three variables simultaneously: pretreatment pH and sodium phosphate concentration, and the concentration of acetic acid (one of the reactants) in the reaction mixture. In only 26 experimental runs, this optimization process determined a combination of variable settings that yielded a reaction product approx 180 times faster than achieved with untreated enzyme. Evidence is presented to demonstrate that locating this optimum with single-factor experiments would be inefficient. This article demonstrates the efficiency of the method of steepest ascent particularly for evaluation of enzymatic reaction conditions exhibiting significant interactions.
Garcia
Rafael A
RA
Department of Agricultural and Biosystems Engineering, The University of Arizona, Tucson, AZ 85721, USA. rgarcia@errc.ars.usda.gov
Riley
Mark R
MR
D019342:Acetic Acid / Q000737:chemistry; D000080462:Acyclic Monoterpenes; D002849:Chromatography, Gas; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D039821:Monoterpenes / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry
D000080462:Acyclic Monoterpenes; D039821:Monoterpenes; D010710:Phosphates; D008049:Lipase; C007078:citronellol; D019342:Acetic Acid; C018279:sodium phosphate