title: A novel method for the purification of porcine phospholipase A2 expressed in E. coli.
abstract: Porcine phospholipaseA2 expressed in E. coli as a fusion protein was isolated, renatured and specifically cleaved by trypsin as described in (1). Active phospholipaseA2, was purified to homogeneity on a column of PBE-94 over a pH region 7.4-4.5. Using this method, several phospholipase A2 mutant enzymes have now been purified in a single step and all behaved identically during chromatofocusing. The method will therefore be extremely useful not only for those interested in understanding the structure-function relationships of phospholipaseA2 but also for preparing the enzyme in large quantities for industrial and pharmaceutical purposes.
authors.affiliation: AFRC Institute of Food Research, Reading Laboratory, Berks, U.K.
authors.forename: K M
authors.initials: KM
authors.lastname: Bhat
authors.forename: I G
authors.initials: IG
authors.lastname: Sumner
authors.forename: B N
authors.initials: BN
authors.lastname: Perry
authors.forename: M E
authors.initials: ME
authors.lastname: Collins
authors.forename: R W
authors.initials: RW
authors.lastname: Pickersgill
authors.forename: P W
authors.initials: PW
authors.lastname: Goodenough
mesh_terms: D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D002479:Inclusion Bodies / Q000201:enzymology; D007700:Kinetics; D008823:Micelles; D008970:Molecular Weight; D010741:Phospholipases A / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D054467:Phospholipases A2; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D013552:Swine; D014357:Trypsin
chemical_list: D008823:Micelles; D011993:Recombinant Fusion Proteins; D010741:Phospholipases A; D054467:Phospholipases A2; D014357:Trypsin