Abstract
This chapter discusses the preparation of porphobilinogen (PBG). PBG is a normal intermediate in the biosynthesis of heme. By using both δ-aminolevulinic acid and the enzyme δ- aminolevulinic acid dehydratase, an enzymatic method has been developed for the preparation of porphobilinogen with good yield. Enzyme is stored as a precipitate at -10°. A solution of 10–20 mg protein per milliliter in 0.134 M Na 2 HPO 4 -KH 2 PO 4 buffer (pH 6.8) is prepared when needed. PBG isolated can be identified by using different methods. By paper chromatography, a biosynthetic PBG runs as a single spot having the same R f as that for the markers. A solution of biosynthetic PBG when mixed with an equal volume of Ehrlich's reagent gives maxima at 555 m μ and 525 m μ. PBG can be enzymatically converted into uroporphyrinogens by means of the deaminase–isomerase enzymatic complex.
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