Biosynthetic studies on tenellin and aminoisobutyrate metabolism in streptomyces sp

Abstract

This thesis is divided into two parts. Part 1 covers the biosynthesis of the fungal metabolite tenellin, and Part 2 the metabolism of β-aminoisobutyrate m Streptomyces sp. Tenellin is a yellow pigment of the fungus Beamaria bassiana. It is of mixed biosynthetic origin, being derived from a polyketide moiety and the amino acid L-phenylalanine. The timing of the C-methylations of the polyketide chain is discussed in Chapter 2, which describes attempts to incorporate deuterium labelled partially assembled putative intermediates into the polyketide. The biosynthesis of the pyridone ring of tenellin requkes the condensation of the polyketide moiety with a rearranged phenylpropanoid unit derived from phenylalanine. The nature of this intriguing intramolecular rearrangement is discussed in Chapters 3 and 4. A phenylalanine derived tetramic acid, proposed as an intermediate in the biosynthesis, has been synthesised, and used in biosynthetic investigations. The results of these investigations and the subsequent identification of tyrosine as a closer precursor to tenellin argue against its intermediacy. The failure of [2-(^13)C(^2)H(^15)N]-phenylalanine to become incorporated intact suggests a transamination process for phenylalanine / tyrosine prior to incorporation. Preliminary investigations suggest para-hydroxy phenyllactate may be die substrate for the rearranging enzyme and a more direct precursor to tenellin. β-Aminoisobutyrate, the end product of reductive thymine catabolism, contributes to both the propionate and butyrate pools in Streptomyces sp. The pathway of incorporation into the isobutyrate / butyrate pool has been investigated, and confirmed to be the reverse of that known to occur in L-valine metabolism. A mutant strain of Streptomyces avemitilis, unable to produce isobutyrate, was used due to low level incorporations into the branched-chain fatty acids. This work was carried out in collaboration with Dr. Hamish McArthur, Pfizer Central Research Division, Groton, USA, and Dr. Kevin Reynolds, Department of Pharmaceutical Science, University of Maryland.