Isolation and molecular characterisation of the benzoate-para-hydroxylase gene (bphA) of Aspergillus niger: A member of a new gene family of the cytochrome P450 superfamily

Gorcom, R.F.M. van; Boschloo, J.G.; A, Kuijvenhoven; Lange, Jacqueline; Vark, Arjen J. van; Bos, C.J.; Balken, J. A. M. van; Pouwels, Peter H.; Hondel, Cees A. M. J. J. van den

doi:10.1007/bf00265053

Cited by 67 publications

(36 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Our results also revealed that A. oryzae produces ferulic acid in the fermentation mixture, a process enhanced by the addition of aromatic compounds to the medium. The enzymes P450 (monooxygenase), ferulate esterase and xylanase, are also induced by aromatic compounds such as benzoic acid, vanillin and coumaric acid 7,21,32 . In the present study, the level of induction of ferulic acid by other aromatic compounds, including vanillic acid and benzoic acid, was similar to the level induced by vanillin (data not shown).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Ferulic Acid Production in the Brewing of Rice Wine (Sake)

Uno

Itoh

Miyamoto³

et al. 2009

Journal of the Institute of Brewing

View full text Add to dashboard Cite

The traditional Japanese alcoholic beverage sake is produced by fermentation of rice by Saccharomyces cerevisiae and Aspergillus oryzae. A. oryzae releases ferulic acid, an antioxidant, from steamed rice during the fermentation process. The concentration of ferulic acid increased with time during fermentation and the production rate peaked 9-12 days post inoculation. Analysis of the fermentation cultures of Aspergillus oryzae, by highperformance liquid chromatography (HPLC), revealed that pcoumaric acid induced an 18.9-fold increase in the level of ferulic acid. Furthermore, SDS-PAGE analysis revealed an increase or decrease in the level of specific proteins after the addition of p-coumaric acid to fermentation cultures of Aspergillus oryzae. Ferulate esterase (FAE) activity was observed in the fermented sake ten days following the start of the fermentation process. These results suggest that the level of ferulic acid is regulated by the enzymes synthesized by A. oryzae during the sake brewing process.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Aromatic compounds induce production of enzymes such as P450, xylase and ferulate esterase 7,21,32 . Vanillin (formed from ferulic acid) and p-coumaric acid are two aromatic compounds that are structurally very similar to ferulic acid 7,11 .…”

Section: Induction Of Ferulic Acid By Aromatic Compoundsmentioning

confidence: 99%

Ferulic Acid Production in the Brewing of Rice Wine (Sake)

Uno

Itoh

Miyamoto³

et al. 2009

Journal of the Institute of Brewing

View full text Add to dashboard Cite

show abstract

“…In addition to phacA, four stc genes of the A. nidulans sterigmatocystin biosynthetic cluster encode CYP enzymes (24). The closely related organism A. niger has a very specific benzoate-4 hydroxylase enzyme, encoded by the bphA gene (25). Notably, PhacA and BphA each define a new CYP family, suggesting that metabolically versatile saprophytic fungi may represent an as yet unexplored source of variability for CYP enzymes catalyzing novel metabolic reactions.…”

Section: Discussionmentioning

confidence: 99%

Disruption of phacA, an Aspergillus nidulans Gene Encoding a Novel Cytochrome P450 Monooxygenase Catalyzing Phenylacetate 2-Hydroxylation, Results in Penicillin Overproduction

Mingot

Peñalva

Fernández-Cañón³

1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

Aspergillus nidulans utilizes phenylacetate as a carbon source via homogentisate, which is degraded to fumarate and acetoacetate. Mutational evidence strongly suggested that phenylacetate is converted to homogentisate through two sequential hydroxylating reactions in positions 2 and 5 of the aromatic ring. Using cDNA substraction techniques, we have characterized a gene, denoted phacA, whose transcription is strongly induced by phenylacetate and which putatively encodes a cytochrome P450 protein. A disrupted phacA strain does not grow on phenylacetate but grows on 2-hydroxy-or 2,5-dihydroxyphenylacetate. Microsomal extracts of the disrupted strain are deficient in the NADPH-dependent conversion of phenylacetate to 2-hydroxyphenylacetate. We conclude that PhacA catalyzes the ortho-hydroxylation of phenylacetate, the first step of A. nidulans phenylacetate catabolism. The involvement of a P450 enzyme in the ortho-hydroxylation of a monoaromatic compound has no precedent. In addition, PhacA shows substantial sequence divergence with known cytochromes P450 and defines a new family of these enzymes, suggesting that saprophytic fungi may represent a source of novel cytochromes P450.Phenylacetate is a precursor for benzylpenicillin production. phacA disruption increases penicillin production 3-5-fold, indicating that catabolism competes with antibiotic biosynthesis for phenylacetate and strongly suggesting strategies for Penicillium chrysogenum strain improvement by reverse genetics.Aerobic degradation of aromatic hydrocarbons by microbes involves the action of oxygenases (enzymes that incorporate one or two atoms from dioxygen into substrates) acting at two different levels in specific catabolic pathways (1, 2). First, oxygenase enzymes acting at the upstream segment of these pathways incorporate one (monooxygenases, aromatic ring hydroxylases) or two (aromatic ring dioxygenases) oxygen atoms into the aromatic substrate as hydroxyl groups, preparing the ring for a subsequent ring-opening step. In this second step, the dihydroxylated aromatic ring is opened by ring-cleavage dioxygenases.Monooxygenases are a mechanistically diverse group of enzymes (1) including, for example, flavoproteins such as p-hydroxybenzoate hydroxylase (3), multicomponent enzymes such as Pseudomonas mendocina toluene 4-monoxygenase, in which one of the terminal hydroxylase polypeptides contains a binuclear iron cluster (4), or heme-containing cytochrome P450 systems. Monooxygenases of the cytochrome P450 superfamily (5, 6) are widely distributed among living organisms and catalyze a multiplicity of biosynthetic and catabolic reactions, usually with narrow substrate specificity, including the hydroxylation of a variety of lipophylic drugs.In common with other saprophytic microbes, the genetically amenable, obligate aerobic fungus Aspergillus nidulans shows notable metabolic versatility. For example, it can use the aromatic hydrocarbon compound phenylacetate (PhAc) 1 as sole carbon source. Despite the abundant information available on the cataboli...

show abstract

“…The MIC of benzoic acid required to inhibit spore germination by A. niger was, as expected since benzoic acid is not a PadA1 substrate, identical in both the mutant and the reference strains (AXP8-1) ( Table 1). The degradation of benzoic acid by A. niger and other fungi has been well documented (18)(19)(20)22) and differs in mechanism from that for breakdown of sorbic and cinnamic acids.…”

mentioning

confidence: 99%