Affinity chromatography of B6-vitamin-dependent enzymes: purification of pig-heart branched-chain amino acid transaminase.
Pig-heart branched-chain amino acid transaminase (EC 2.6.1.42) was purified to near homogeneity with a yield of 27%. A prepurification was performed by heat treatment, gel chromatography and DEAE-Sepharose methods. For the final step, several affinity gels were tested and the one containing cycloserine coupled to CNBr-activated Sepharose 4B was selected. This effected an additional five-fold purification with a yield of 60%. The present affinity results are compared with corresponding studies with other aminotransferases in an attempt to find possible universal techniques for their purification.
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D009206:Myocardium / Q000201:enzymology*; D011506:Proteins / Q000032:analysis; D011736:Pyridoxine / Q000378:metabolism*; D013552:Swine; D000637:Transaminases / Q000032:analysis*
D011506:Proteins; D000637:Transaminases; C019798:branched-chain-amino-acid transaminase; D011736:Pyridoxine