Print Email Facebook Twitter Isolation and molecular characterisation of the benzoate-para-hydroxylase gene (bphA) of Aspergillus niger Title 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 Author van Gorcom, R.F.M. Boschloo, J.G. Kuijvenhoven, A. Lange, J. van Vark, A.J. Bos, C.J. van Balken, J.A.M. Pouwels, P.H. van den Hondel, C.A.M.J.J. Publication year 1990 Abstract The gene coding for benzoate-para-hydroxylase (bphA) of Aspergillus niger was cloned using differential hybridisation techniques and complementation of mutants deficient in this enzyme activity. The nucleotide sequence of the gene was determined, the presence of two introns was shown and the transcription start and termination sites were determined. The structure of the mRNA upstream from the long open reading frame (ORF) is unusual. It contains two small, overlapping ORFs whose function is unknown. Comparison of the deduced amino acid sequence of the protein with the sequences present in the databanks, indicated a significant similarity of BPH to the superfamily of cytochrome P450 enzymes. Further analysis revealed that this protein is a member of a new p450 gene family designated P450LIII. The gene is designated CYP53. To increase the BPH activity of A. niger, multiple copies of the bphA gene were introduced into the genome of a recipient strain by transformation. Although increased intracellular levels of the BPH protein could be detected, the BPH enzyme activity was decreased, suggesting titration of another essential component. Subject ComplementationMonooxygenaseOverexpressionRecombinant DNAUpstream reading framesCytochrome p450Unspecific monooxygenaseAspergillus nigerCell cultureFungusGenetic engineeringGenetic recombinationHeredityNonhumanPriority journalAmino Acid SequenceAspergillus nigerBase SequenceBlotting, WesternCloning, MolecularCytochrome P-450 Enzyme SystemGene Expression Regulation, FungalGenes, FungalGenetic Complementation TestIntronsMolecular Sequence DataMultigene FamilyMutationOxygenasesRestriction MappingSequence Homology, Nucleic AcidTranscription, GeneticAspergillus nigerFungi To reference this document use: http://resolver.tudelft.nl/uuid:8ff26a33-d2a6-4548-bb7e-ac3fa1bc23e5 TNO identifier 231247 ISSN 0026-8925 Source Molecular and General Genetics, 223 (223), 192-197 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.