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
Complementation
Monooxygenase
Overexpression
Recombinant DNA
Upstream reading frames
Cytochrome p450
Unspecific monooxygenase
Aspergillus niger
Cell culture
Fungus
Genetic engineering
Genetic recombination
Heredity
Nonhuman
Priority journal
Amino Acid Sequence
Aspergillus niger
Base Sequence
Blotting, Western
Cloning, Molecular
Cytochrome P-450 Enzyme System
Gene Expression Regulation, Fungal
Genes, Fungal
Genetic Complementation Test
Introns
Molecular Sequence Data
Multigene Family
Mutation
Oxygenases
Restriction Mapping
Sequence Homology, Nucleic Acid
Transcription, Genetic
Aspergillus niger
Fungi
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TNO identifier
231247
ISSN
0026-8925
Source
Molecular and General Genetics, 223 (223), 192-197
Document type
article