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Applied and Environmental Microbiology, April 2000, p. 1517-1522, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Novel Scheme for Biosynthesis of Aryl Metabolites from L-Phenylalanine in the Fungus Bjerkandera adusta

Carmen Lapadatescu,1 Christian Giniès,1 Jean-Luc Le Quéré,1 and Pascal Bonnarme2,*

Laboratoire de Recherches Sur les Arômes (LRSA), Institut National de la Recherche Agronomique, 21034 Dijon cedex,1 and Laboratoire de Génie et Microbiologie des Procédés Alimentaires (LGMPA), CBAI, Institut National de la Recherche Agronomique, 78850 Thiverval-Grignon,2 France

Received 27 October 1999/Accepted 18 January 2000

Aryl metabolite biosynthesis was studied in the white rot fungus Bjerkandera adusta cultivated in a liquid medium supplemented with L-phenylalanine. Aromatic compounds were analyzed by gas chromatography-mass spectrometry following addition of labelled precursors (14C- and 13C-labelled L-phenylalanine), which did not interfere with fungal metabolism. The major aromatic compounds identified were benzyl alcohol, benzaldehyde (bitter almond aroma), and benzoic acid. Hydroxy- and methoxybenzylic compounds (alcohols, aldehydes, and acids) were also found in fungal cultures. Intracellular enzymatic activities (phenylalanine ammonia lyase, aryl-alcohol oxidase, aryl-alcohol dehydrogenase, aryl-aldehyde dehydrogenase, lignin peroxidase) and extracellular enzymatic activities (aryl-alcohol oxidase, lignin peroxidase), as well as aromatic compounds, were detected in B. adusta cultures. Metabolite formation required de novo protein biosynthesis. Our results show that L-phenylalanine was deaminated to trans-cinnamic acid by a phenylalanine ammonia lyase and trans-cinnamic acid was in turn converted to aromatic acids (phenylpyruvic, phenylacetic, mandelic, and benzoylformic acids); benzaldehyde was a metabolic intermediate. These acids were transformed into benzaldehyde, benzyl alcohol, and benzoic acid. Our findings support the hypothesis that all of these compounds are intermediates in the biosynthetic pathway from L-phenylalanine to aryl metabolites. Additionally, trans-cinnamic acid can also be transformed via beta -oxidation to benzoic acid. This was confirmed by the presence of acetophenone as a beta -oxidation degradation intermediate. To our knowledge, this is the first time that a beta -oxidation sequence leading to benzoic acid synthesis has been found in a white rot fungus. A novel metabolic scheme for biosynthesis of aryl metabolites from L-phenylalanine is proposed.


* Corresponding author. Mailing address: Laboratoire de Génie et Microbiologie des Procédés Alimentaires (LGMPA), CBAI, Institut National de la Recherche Agronomique, 78850 Thiverval-Grignon, France. Phone: 331-3081 5388. Fax: 331-3081 5597. E-mail: bonnarme{at}platon.grignon.inra.fr.


Applied and Environmental Microbiology, April 2000, p. 1517-1522, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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