title: Intramolecular esterification by lipase powder in microaqueous benzene: factors affecting activity of pure enzyme.
abstract: Various factors affecting the catalytic activity of pure lipase of Pseudomonas fluorescens in microaqueous benzene were investigated with respect to lactonization of 15-hydroxypentadecanoic acid. Without deposition of the enzyme or of the enzyme plus activity enhancer (additive) on celite powder, the pure enzyme was very poorly dispersed in the microaqueous benzene, resulting in very low activity. The enzyme immobilized on celite powder exhibited the highest activity at a free water content of ca. 0.083%. When a sugar alcohol such as erythritol, arabitol, or sorbitol was added before lyophilization with approximate proportion of 3 g/g enzyme, marked increases in the enzyme activity were observed at a shifted optimal free water content, i.e., 0.04%. Inclusion of phosphotidylcholine resulted in a somewhat higher activity than in the system of enzyme plus celite only. Addition of lactose, bovine serum albumin, casein, dextran, polyvinyl alcohol, phosphate, or NaCl all caused a decrease in the enzyme activity. From the effects of the additives examined, it is deduced that the following three factors are required for a pure enzyme to exhibit its full activity in a water-immiscible organic solvent: (1) optimum moisture content, (2) disperser (support particles having enough surface area on which the enzyme is thinly deposited), and (3) activity enhancer (additive) at optimum concentration The importance of noting the purity of the enzyme preparation is emphasized when its catalysis in an organic solvent is investigated.
authors.affiliation: Laboratory of Bioreaction Engineering, Department of Food Science and Technology, School of Agriculture, Nagoya University, Chikusa-ku, Nagoya 464-01, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Yamane
authors.forename: T
authors.initials: T
authors.lastname: Ichiryu
authors.forename: M
authors.initials: M
authors.lastname: Nagata
authors.forename: A
authors.initials: A
authors.lastname: Ueno
authors.forename: S
authors.initials: S
authors.lastname: Shimizu