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Appl Environ Microbiol. 1967 January; 15(1): 110-113
Copyright © 1967 American Society for Microbiology. All Rights Reserved.

Instrumental Assay of Microbial Lipase at Constant pH¹

Department of Microbiology and Public Health, Michigan State University, East Lansing, Michigan

ABSTRACT

A rapid, accurate method with high sensitivity and reproducibility, and having the advantage of a short incubation period under constant pH, has been developed for routine measurement of microbial lipase. Assembled from readily available and economical instrumental components, the apparatus includes a pH meter, a thermoelectric heating and stirring device, a motor-driven burette, and an automatic recorder. The reaction mixture, consisting of 5 ml of a 10% olive oil-gum arabic emulsion, 2 ml of 3 M NaCl, 2 ml of sodium taurocholate (15 mg/ml) of 0.075 M CaCl₂, 5 ml of water, and 1 ml of enzyme solution, was adjusted to pH 8.0 and 37 C. The pH was maintained at a constant value by automatic addition of 0.01 N NaOH during the incubation period, which usually lasted 5 min. A lipase unit, derived from the use of this technique, may be defined as the number of microequivalents of acid liberated per minute under the specified conditions. The method was sensitive to 0.01 units. Various organisms tested produced 0.17 to 1.32 units per ml of the cell filtrate. An Arrhenius plot for staphylococcal lipase yielded 14,500 cal for function A (energy of activation).

¹ Published with the approval of the Director of the Michigan Agricultural Station as Journal Article 3907.

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