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Restricted activation of general amino acid control under conditions of glutamine limitation in Neurospora crassa.

In Neurospora crassa limitation for single amino acids normally results in increased formation of enzymes required for amino acid synthesis via 'general amino acid control'. Glutamine limitation, however, led to comparatively low and delayed derepression of enzyme synthesis. Nitrate reductase activity increased steeply under these conditions confirming that de novo protein synthesis could occur. Derepression levels were unaffected by addition of glutamine-derived metabolites. Only small and delayed increases in mRNA levels occurred for the anabolic enzyme genes arg-12, his-3 and trp-1 under conditions of glutamine limitation in contrast to the immediate and far larger increase found on histidine limitation. The trans-acting regulatory gene of general amino acid control in Neurospora, cpc-1, responded with a significant increase in mRNA level to histidine and to glutamine limitation. The restricted response of the amino acid synthesis genes could imply a post-transcriptional block to the positive regulatory function of cpc-1 under condition of glutamine limitation. The results suggest that the expression of general amino acid control is restricted under conditions of inadequate nitrogen supply.

Alcohol Oxidoreductases↗

Patterns of metabolites produced from the fluoroquinolone enrofloxacin by basidiomycetes indigenous to agricultural sites.

Supernatants of mycelial cultures of seven basidiomycetous fungi indigenous to agricultural sites were evaluated for metabolites generated from the veterinary fluoroquinolone enrofloxacin (EFL) by employing high-performance liquid chromatography/high-resolution electrospray ionization mass spectrometry. From exact masses, molecular formulae were derived, and the most probable chemical structures were deduced. Patterns of major metabolites were surprisingly similar but differed greatly from that provided by Gloeophyllum striatum due to the absence of monohydroxylated EFL congeners and a greater variety of metabolites with a modified piperazine moiety. The structures of three metabolites were elucidated by 1H-nuclear magnetic resonance spectroscopy. Of 61 compounds detected, 48 were new, while 13 were known from a pattern of 87 EFL metabolites identified for G. striatum. Ethylpiperazine moieties carrying oxido, hydroxy, oxo, and acetoxy groups, or showing partial degradation, were linked to the unmodified, oxidatively decarboxylated, or multiply hydroxylated core of EFL and to isatin- and anthranilic acid-type EFL congeners. Cleavage of the fluoro-aromatic bond was observed for two, 14CO2 formation for six species. Metabolites with a hydroxylated aromatic part implied subsequent ring cleavage to be brought about by the formation of potentially four oxidizable ortho-aminophenol- and one catechol-type intermediates. EFL degradation appears to be a common activity among basidiomycetes.

Agriculture↗

Parallel synthesis and in vitro activity of novel anthranilic hydroxamate-based inhibitors of the prostaglandin H2 synthase peroxidase activity.

Currently available non-steroidal anti-inflammatory drugs (NSAIDs) such as aspirin are directed at the cyclooxygenase (COX) site, but not the peroxidase (POX) activity of prostaglandin H2 synthase (PGHS). They are thus unable to inhibit the free-radical induced tissue injury associated with PGHS peroxidase activity, which can occur independently of the COX site. A lead compound, anthranilic hydroxamic acid (AHA) was found to have significant PGHS-POX inhibitory activity (IC50= 72 microM). To define the critical parameters for PGHS-POX inhibition, we investigated 29 AHA derivatives, synthesised from their acid precursors, using solid phase synthesis. In vitro analysis demonstrated a ten-fold improvement in inhibition with 3,5-diiodoanthranilic hydroxamic acid (IC50= 7 microM).

Enzyme Activation↗