Insoluble material was removed by centrifugation
at 10,000 rpm during 10 min. The supernatant (¡«15 ml)
was applied to a Sephacryl S-200 (1.8
¡Á
100 cm) gel filtration column
(Amersham Biosciences) equilibrated in buffer A. The elution
of the lipase was performed with the same buffer at a flow rate of
18 ml/h.