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Summary: Rhodanese from Thiobacillus A2 was shown by proton nuclear magnetic resonance (NMR) spectroscopy to use dihydrolipoate or dihydrolipoamide as acceptor of the sulphane moiety of thiosulphate with the formation of α-lipoate or lipoamide respectively. Correlation is shown between assays of the enzyme activity by NMR spectroscopy and by ultraviolet spectrophotometry.