Purification of lipase from Chromobacterium viscosum by chromatography on palmitoyl cellulose.
Palmitoyl cellulose was used to adsorb the extracellular lipase [triacylglycerol acyl-hydrolase EC 3.1.1.3] of Chromobacterium viscosum from crude enzyme solution, and the adsorbed enzyme was eluted with a suitable detergent, such as Adekatol 45-S-8 or Triton X-100. The enzyme was then purified by chromatography on a palmitoylated gauze column with an overall recovery of 71% and an increase in the specific activity of 11-fold from the supernatant fluid of bacterial cultures. Further purification procedures included fractionation with acetone, and chromatography on Sephadex G-150 and G-75 columns. Two isoenzymes were obtained, each in a homogeneous state on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis: one had a molecular weight of 120,000 and pI of 3.7 and the other a molecular weight of 30,000 with a pI of 7.3.
D002482:Cellulose; D002846:Chromatography, Affinity; D002861:Chromobacterium / Q000201:enzymology*; D003902:Detergents; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008049:Lipase / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D010169:Palmitic Acids; D013329:Structure-Activity Relationship
D003902:Detergents; D007527:Isoenzymes; D010169:Palmitic Acids; D002482:Cellulose; D008049:Lipase