Molecular characterisation of formamidase from Methylophilus methylotrophus

Eur J Biochem. 1996 Sep 1;240(2):314-22. doi: 10.1111/j.1432-1033.1996.0314h.x.


A 3.2-kbp PstI fragment of DNA encoding formamidase from the methylotrophic bacterium Methylophilus methylotrophus which had previously been cloned (pNW3) [Wyborn, N.R., Scherr, D.J. & Jones, C.W. (1994) Microbiology 140, 191-195], was subcloned as a 2.3 kbp HindIII fragment (pNW323). Nucleotide sequencing showed that the subclone contained two genes which encoded formamidase (fmdA) and a possible regulatory protein (fmdB). Predicted molecular masses for FmdA and FmdB were 44438 Da (compared with approximately 44500 Da by electrospray mass spectrometry and 51000 Da by SDS/PAGE of the purified enzyme) and 12306 Da, respectively. The derived amino acid sequence of formamidase was supported by N-terminal amino acid sequencing of the enzyme and of proteolytic fragments prepared from it using V8 endoproteinase and was 57% similar to that of the acetamidase from Mycobacterium smegmatis. The structural similarities between these two enzymes, and their existence as a separate class of bacterial amidase, were confirmed by immunological investigations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / chemistry*
  • Amidohydrolases / classification
  • Amidohydrolases / genetics
  • Amidohydrolases / metabolism
  • Amino Acid Sequence
  • Ammonia / pharmacology
  • Blotting, Western
  • Cloning, Molecular
  • Codon
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Gene Expression
  • Genes, Bacterial
  • Gram-Negative Aerobic Bacteria / enzymology*
  • Mass Spectrometry
  • Molecular Sequence Data
  • Molecular Weight
  • Open Reading Frames / genetics
  • Restriction Mapping
  • Sequence Alignment
  • Sequence Analysis
  • Zinc Fingers


  • Codon
  • Ammonia
  • Amidohydrolases
  • formamidase

Associated data

  • GENBANK/U09411
  • GENBANK/X99632