Part:BBa_K3599005

pfNIT

Overview

Nitrilase coming from Nectria haematococca, catalysis of the reaction: nitrile + 2H₂O = carboxylate + NH₃. The codon usage was optimized for expression in Escherichia coli.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Primitive Species Background

Pseudomonas fluorescens are Gram-negative rod shaped bacteria that inhabit soil, plants, and water surfaces. The optimum growth temperature is between 25-30 degrees Celsius. The Pf-5 strain resides in the plant’s rhizosphere and produces a variety of secondary metabolites including antibiotics against soil borne plant pathogens. Pseudomonas fluorescens PFO-1 is well adapted to the soil where it was first isolated in agricultural soil. Pseudomonas fluorescens strain SBW25 grow on plant leaves and roots where they can contribute to plant growth. Soluble, green fluorescent pigments are produced when the iron concentration is low. The significance of these organisms have increased because of their ability to degrade various pollutants and their use as bio-control against pathogens. Sequencing the genome provided further information of its environmental interaction ands its metabolic capabilities, which can be used against agricultural disease control. Pseudomonas fluorescens is interesting and important to study because it produces a particular antibiotic (Mupirocin) which has been proven effective in treating certain kinds of skin, ear, and eye disorders.

Design & Experience

In iGEM2020 TPR_China Project, the coding sequence of pfNIT were founded and optimized, further characterization of its enzymatic activity could be done by iGEM teams in the fature.

Reference

Kiziak, Christoph, Conradt, Doris, Stolz, & Andreas, et al. (2005). Nitrilase from pseudomonas fluorescens ebc191: cloning and heterologous expression of the gene and biochemical characterization of the recombinant enzyme. Microbiology (13500872). 10.1099/mic.0.28246-0