Journal of Lipid Research, Vol 21, 180-185, Copyright © 1980 by Lipid Research, Inc.
Purification of phospholipase B from Penicillium notatum by hydrophobic chromatography on palmitoyl cellulose
S Imamura and Y Horiuti
Phospholipase B (lysolecithin acyl-hydrolase, EC 3.1.1.5) from the mycelia
of Penicillium notatum (Institute for Fermentation, Osaka, Japan; No.4640)
was adsorbed from a crude solution to palmitoyl cellulose. Adsorption was
efficient at pH 4 at low ionic strength (10 mM buffer), and at pH 4-9 at
higher ionic strength (1-2M NaCl in 10 mM buffer). The adsorbed enzyme was
eluted from the cellulose with a suitable detergent, such as Adekatol
SO-120, Triton X-100, or deoxycholate. As an application of this procedure,
the enzyme was purified from an extract of the mycelia by column
chromatography on a palmitoylated textile (palmitoylated gauze) with an
overall recovery of 59% and a 38-fold increase in specific activity. By
subsequent column chromatographies on Amberlite XAD-2, Sephadex G-100 and
G-150, and DEAE- Sephadex A-50, the enzyme was purified about 4,000-fold to
a nearly homogeneous state from a mycelium extract with an overall recovery
of 37%.
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