Relative effectiveness of pretreatments on performance of Rhizomucor miehei lipase in nonpolar reaction media.
Enzymes can be used in nonpolar reaction media to modify water-insoluble substrates. A variety of pretreatments, applied to the enzyme prior to introduction to the nonpolar media, can improve enzyme activity. However, the various pretreatments have not been studied using directly comparable conditions, nor have they been applied simultaneously to test for interactive effects. This work evaluates pretreatment of lipase with various classes of additives. The pretreated lipase is used to catalyze esterification between citronellol and acetic acid in a medium of n-hexane. The effectiveness of a particular pretreatment is presented in terms of relative performance (RP), which is equal to the number of times faster the pretreated lipase catalyzes the reaction relative to untreated lipase. The individual and interactive effects of the pretreatment factors were studied and compared. Buffer salts had a much stronger performance-enhancing effect than nonbuffer salts; pretreatment with 90% (w/w) sodium phosphate yielded lipase with an RP of approx 64. A strong interaction was found between the treatments with sodium phosphate and pH adjustment. These treatments may mitigate the inhibitory effect of acetic acid. Activating effects of phase interfaces and active-site protectants are shown to be complementary to other treatments, demonstrating that they likely act by distinct mechanisms.
Garcia
Rafael A
RA
Department of Agricultural and Biosystems Engineering, Shantz Building, The University of Arizona, Tucson, AZ 85721, USA. rgarcia@errc.ars.usda.gov
Riley
Mark R
MR
D019342:Acetic Acid / Q000138:chemical synthesis / Q000737:chemistry; D000080462:Acyclic Monoterpenes; D001665:Binding Sites; D002384:Catalysis; D004789:Enzyme Activation; D005069:Evaluation Studies as Topic; D006586:Hexanes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000737:chemistry*; D039821:Monoterpenes / Q000138:chemical synthesis / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry; D020103:Rhizomucor / Q000201:enzymology*; D013499:Surface Properties; D014867:Water / Q000737:chemistry
D000080462:Acyclic Monoterpenes; D006586:Hexanes; D039821:Monoterpenes; D010710:Phosphates; D014867:Water; C026385:n-hexane; D008049:Lipase; C007078:citronellol; D019342:Acetic Acid; C018279:sodium phosphate