Delta 1-pyrroline-5-carboxylate reductase from baker's yeast: further purification by affinity chromatography with 5' AMP-Sepharose 4B.
A homogeneous preparation of delta 1-pyrroline-5-carboxylate reductase (L-proline: NAD (P)+ 5-oxidoreductase, EC 1.5.1.2) was obtained from baker's yeast by an affinity chromatography, using 5' AMP-Sepharose 4B. After the 1st DEAE-Sephadex column chromatography, the enzyme absorbed on 5' AMP-Sepharose column was eluted with 1 mM ATP. The final preparation was homogeneous on polyacrylamide gel electrophoresis and purified 3500-fold from the crude extract. This purified enzyme was very useful as a coupling enzyme for the assays of ornithine and pyrroline-5-carboxylate in tissues, and for the rate assay of ornithine aminotransferase activity.
D000249:Adenosine Monophosphate; D002846:Chromatography, Affinity; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000302:isolation & purification*; D011762:Pyrroline Carboxylate Reductases / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012685:Sepharose
D000249:Adenosine Monophosphate; D012685:Sepharose; D000587:Oxidoreductases Acting on CH-NH Group Donors; D011762:Pyrroline Carboxylate Reductases