[16]l-3-hydroxyacyl coenzyme A dehydrogenase from pig heart muscle

摘要: Publisher Summary This chapter discusses the determination of L-3-hydroxyacyl coenzyme A dehydrogenase from pig heart muscle. mitochondrial enzyme participates in β-oxidation fatty acids. It can be distinguished acetoacetyl-CoA reductase (D-3-hydroxyacyl-CoA : NADP oxidoreductase) by stereochemical configuration hydroxyacyl substrate and principal cofactor utilized. The activity is routinely measured monitoring decrease absorption reduced nicotinamide adenine dicucleotide (NADH) at 340 nm, which accompanies conversion S-acetoacetyl ester to corresponding β-hydroxy compound. Either S-acetoacetylpantetheine or S-acetoacetyl-CoA used as substrate. However, because expense more pronounced product inhibition encountered with CoA ester, choice. purification scheme described carried out 80 hearts (20 kg). initial stages are best performed batches 12–15 hearts. All buffers contained 1 m M EDTA 2 β-mercaptoethanol, entire preparation was 0–4°. Specific activities based on protein determined microbiuret reaction.