Lipase made active in hydrophobic media by coupling with polyethylene glycol

Abstract

AbstractLipases from various microorganisms were chemically modified with polyethylene glycol derivatives: 2,4‐bis[O‐methoxypoly(ethylene glycol)]‐6‐chloro‐s‐triazine (activated PEG2, a chain‐shaped polymer) and copolymer of polyoxyethylene allyl methyl diether and maleic anhydride (activated PM, a comb‐shaped polymer). Because each polymer is amphipathic, the modified lipases become soluble not only in aqueous solution but also in hydrophobic media. They exhibit potent catalytic actions for ester synthesis and ester exchange reactions, the reverse reaction of hydrolysis, in transparent organic solvents and also in oily substrates without organic solvents. With PEG2‐lipases, macrocyclic lactone and gefarnate (geranyl farnesylacetate) were synthesized in high yields from 16‐hydroxy‐hexadecanoic acid ethyl ester and from farnesylacetic acid and geraniol in organic solvents, respectively. The modified lipase catalyzed the esterification preferentially with the (R)‐isomer of secondary alcohols. Because the ester synthesis reactions with modified lipase proceeded in the transparent benzene system, the kinetic parameters (Michaelis constant and maximum velocity) were obtained by reciprocal plotting according to the Michaelis equation. With comb‐shaped polymer as modifier, PM‐lipase catalyzed effectively the reverse reaction of hydrolysis in organic solvents. The properties of each modified lipase are discussed in relation to those of the nonmodified lipase.