Part:BBa_K3599003

adNIT

Overview

Nitrilase coming from Achromobacter denitrificans, catalysis of the reaction: a nitrile + 2H₂O = a 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
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 629
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 191
  Illegal BsaI site found at 374
  

Primitive Species Background

The type species of gram negative, aerobic bacteria in the genus. A genus of strictly aerobic, non-spore forming rods. Soil and water are regarded as the natural habitat. They are sometimes isolated from a hospital environment and humans. Previously in the genus Alcaligenes, the classification and nomenclature of this species has been frequently emended. The two subspecies, Achromobacter xylosoxidans subsp. denitrificans and Achromobacter xylosoxidans subsp. xylosoxidans are associated with infections.

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_K3599009.

Reference

Novikov, A. D. , Riabchenko, L. E. , Leonova, T. E. , Larikova, G. A. , & Yanenko, A. S. . (2016). A bacterial strain alcaligenes denitrificans С-32 contains two nitrilases with different substrate specificities. 10.21519/0234-2758-2016-32-6-45-52