Synergistic Actions of Tailoring Enzymes in Pradimicin Biosynthesis

Three key tailoring enzymes in pradimicin biosynthesis: PdmJ, PdmW, and PdmN, were investigated. PdmW was characterized as the C‐6 hydroxylase by structural characterization of the corresponding product, 6‐hydroxy‐G‐2A. The efficiencies of the C‐5 and C‐6 hydroxylations, catalyzed respectively by PdmJ and PdmW, were low when they were expressed individually with the early biosynthetic enzymes that form G‐2A. When these two cytochrome P450 enzymes were co‐expressed, a dihydroxylated product, 5,6‐dihydroxy‐G‐2A, was efficiently produced, indicating that these two enzymes work synergistically in pradimicin biosynthesis. Heterologously expressed PdmN in Streptomyces coelicolor CH999 converted G‐2A to JX137a by ligating a unit of D ‐alanine to the carboxyl group. PdmN has relaxed substrate specificity toward both amino acid donors and acceptors. Through combinatorial biosynthesis, a series of new pradimicin analogues were produced.     

A testosterone metabolite is rewarding to ovariectomized female rats.

Anabolic androgenic steroids have become a major class of drugs of abuse among a growing population of male and female adolescents. Although the rewarding and reinforcing properties of androgens have been demonstrated in male rodents, it is unknown whether these properties are apparent in female rats. In this study, conditioned place preference and self-administration paradigms showed that the endogenous androgen metabolite 3αDIOL is rewarding and reinforcing in ovariectomized female rats. Because 3αDIOL can be synthesized de novo in the brain, it is hypothesized that this neurosteroid provides a permissive neurochemical environment that modulates reward processes. (PsycINFO Database Record (c) 2010 APA, all rights reserved)