Part:BBa_K3599002

nhNIT

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 39
  
• 1000
  COMPATIBLE WITH RFC[1000]

Primitive Species Background

The fungus Nectria haematococca Mating Population VI (MPVI), also commonly referred to by its asexual name Fusarium solani, is a member of an evolutionary group called the Fusarium solani species complex, which is comprised of about 50 species. The term "mating population" refers to the fact that members of MPVI are sexually fertile with one another, indicating that they form a biological species.
Members of the F. solani species complex have an ubiquitous distribution and colonize a wide variety of habitats. They can be found as soil-inhabiting saprophytes, rhizosphere colonizers, or as pathogens, which cause disease on many plant and animal species, including humans. Perhaps because of their cosmopolitan nature these fungi have an impressive range of metabolic capabilities; they can degrade a variety of recalcitrant compounds such as lignin and lignocellulose, hydrocarbons (e.g., pyrene, fluoroacetate), plastics, pesticides (e.g., DDT, butachlor, ioxynil), and cyanide complexes. Indeed, N. haematococca MPVI is tolerant to many compounds shown to be toxic to other fungi, including antibiotics, heavy metals and metabolic poisons.

Design & Experience

In iGEM2020 TPR_China Project, the nhNIT was expressed using pTac system, which was pre-embedded into commercial expression vector pET-28b. The whole expression cassette can be found in BBa_K3599007.

Reference

Alicja, B., Veselá, Alena, Petříčková, & Philip, et al. (2013). Heterologous expression, purification and characterization of arylacetonitrilases from nectria haematococca and arthroderma benhamiae. Biocatalysis & Biotransformation. 10.3109/10242422.2012.758117
Coleman JJ, Rounsley SD, Rodriguez-Carres M, Kuo A, Wasmann CC, Grimwood J, et al. (2009) The Genome of Nectria haematococca: Contribution of Supernumerary Chromosomes to Gene Expansion. PLoS Genet 5(8): e1000618. 10.1371/journal.pgen.1000618