Part:BBa_K3381005

Mst-CopC-CBM2a

Mst-CopC-CBM2a is a fusion protein consisting of Mst-CopC, a copper-binding domain, as well as CBM2a, a cellulose-binding domain. A linker sequence that was found to naturally occur in a CBM2a fusion was used as a linker domain between Mst-CopC and CBM2a (Courtade, 2018). Additionally, a sequence for a His-tag as well as a TEV recognition sequence was added to the N-terminus for ease of purification (by immobilized-metal affinity chromatography [IMAC]). Fusion proteins containing cellulose-binding modules (CBMs) have readily been used in industrial purification processes, where the CBMs act as affinity tags. Strikingly enough, waste processing seems to be an application that has not been as fervently explored (Zhou, 2020). Furthermore, current waste processing methods fail to provide a way to recover metal ions once extracted, leaving the playing field open to technologies that make metal recovery possible. Mst-CopC-CBM2a is a fusion protein that allows for extraction and recovery of copper(II) from aqueous waste. It binds irreversibly to cellulose, and can also bind Cu(II) (Koropatkin, 2007). This makes it appropriate for affinity chromatography or related uses. Source of this part: Sources of DNA sequences, in order of occurrence in the fusion protein from N' to C': His-tag and TEV linker sequence: Retrieved from His-tag and TEV linker that occurs in bacterial expression vector pMCSG7: https://plasmid.med.harvard.edu/PlasmidRepository/file/sequence/pMCSG7.gb

References: Courtade, G., Forsberg, Z., Heggset, E. B., Eijsink, V. G., & Aachmann, F. L. (2018, August 24). The carbohydrate-binding module and linker of a modular lytic polysaccharide monooxygenase promote localized cellulose oxidation. Retrieved September 30, 2020, from https://www.jbc.org/content/293/34/13006.full

Koropatkin, N., Randich, A. M., Bhattacharyya-Pakrasi, M., Pakrasi, H. B., & Smith, T. J. (2007, September 14). The Structure of the Iron-binding Protein, FutA1, from Synechocystis 6803. Retrieved September 30, 2020, from https://www.jbc.org/content/282/37/27468.full

Zhou, J., Chen, J., Zhuang, N., Zhang, A., Chen, K., Xu, N., . . . Jiang, M. (2020, May 12). Immobilization and Purification of Enzymes With the Novel Affinity Tag ChBD-AB From Chitinolyticbacter meiyuanensis SYBC-H1. Retrieved September 30, 2020, from https://www.frontiersin.org/articles/10.3389/fbioe.2020.00579/full

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 785
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]