Horiuti Y, Imamura S
J Biochem. 1977 Jun;81(6):1639-49. doi: 10.1093/oxfordjournals.jbchem.a131623.
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.
用棕榈酰纤维素从粗酶溶液中吸附粘质色杆菌的胞外脂肪酶[三酰甘油酰基水解酶，EC 3.1.1.3]，并用合适的去污剂（如Adekatol 45-S-8或 Triton X-100）洗脱吸附的酶。然后通过在棕榈酰化纱布柱上进行色谱法对该酶进行纯化，总回收率为71%，比活性比细菌培养物的上清液提高了11倍。进一步的纯化步骤包括用丙酮分级分离，以及在Sephadex G-150和G-75柱上进行色谱法。获得了两种同工酶，在十二烷基硫酸钠（SDS）-聚丙烯酰胺凝胶电泳上均呈均一状态：一种分子量为120,000，pI为3.7，另一种分子量为30,000，pI为7.3。
