Patulin biosynthesis: epoxidation of toluquinol and gentisyl alcohol by particulate preparations from Penicillium patulum.
A crude extract that catalyzes the epoxidation of toluquinol and gentisyl alcohol was isolated from cultures of Penicillium patulum. About 60% of the activity sedimented from crude extract upon centrifugation at 105,000g for 2 h, and at 30,000g for 30 min after precipitation with 30% ammonium sulfate and resuspension in buffer. The quinone epoxide phyllostine, a product of gentisyl alcohol epoxidation, has previously been shown to be an intermediate in the biosynthesis of patulin [Sekiquchi, J., & Gaucher, G. M. (1978) Biochemistry 17, 1785-1791] and was shown to be further converted to neopatulin by the extract. The epoxide product of toluquinol, desoxyphyllostine (2-methyl-5,6-epoxy-1,4-benzoquinone), has not been reported previously from fungal cultures. Its structure was confirmed by GC-mass spectrometry and proton and 13C NMR. Its CD spectrum showed the same shape and signs as that of phyllostine, indicating that it too is an enzymatic product with a similar absolute configuration. Whereas chemical epoxidation of toluquinone and gentisyl quinone occurs with hydrogen peroxide, the enzymatic epoxidation utilized oxygen and the hydroquinone. The epoxidation was inhibited by 1,10-phenanthroline, EDTA, and p-(chloromercuri)benzenesulfonic acid and by degassing with nitrogen, but no inhibition was observed with KCN, catalase, or CO. The apparent Km's were similar for the two substrates (0.17 mM for toluquinol, 0.24 mM for gentisyl alcohol), with both substrates showing inhibition at 1.0 mM. The rate of desoxyphyllostine formation was more than 10 times that of phyllostine formation at equivalent substrate concentrations. Gentisaldehyde was not a substrate for the enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)
Priest
J W
JW
Department of Chemistry, Florida State University, Tallahassee 32306.
Light
R J
RJ
D001592:Benzyl Alcohols / Q000378:metabolism* / Q000494:pharmacology; D001593:Benzyl Compounds / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D004852:Epoxy Compounds / Q000378:metabolism*; D004988:Ethers, Cyclic / Q000378:metabolism*; D006873:Hydroquinones / Q000378:metabolism* / Q000494:pharmacology; D015394:Molecular Structure; D010088:Oxidoreductases / Q000378:metabolism*; D010365:Patulin / Q000096:biosynthesis*; D010407:Penicillium / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D011714:Pyrans; D010837:Vitamin K 1 / Q000031:analogs & derivatives* / Q000378:metabolism
D001592:Benzyl Alcohols; D001593:Benzyl Compounds; D004852:Epoxy Compounds; D004988:Ethers, Cyclic; D006873:Hydroquinones; D011480:Protease Inhibitors; D011714:Pyrans; C016186:vitamin K1 oxide; C062397:2-methyl-1,4-hydroquinone; D010837:Vitamin K 1; D010365:Patulin; D010088:Oxidoreductases; C023111:epoxidase; C004771:2,5-dihydroxybenzyl alcohol