Part:BBa_K4275040

pLAC4-KmarxMFα-TrEGIII-t-tTDH1

The composite part is composed of Kluyveromyces marxianus mating factor alpha (BBa_K4275000) and endoglucanase TrEGIII-t fused with type 1 dockerin (BBa_K4275006).

Kluyveromyces marxianus mating factor alpha (BBa_K4275000) is fused for the purpose of secretion of target enzyme TrEGIII-t, which would then be anchored on CipA1B2C scaffold protein by type I cohesin-dockerin interaction and assembled into the cellulosome complex.

The part is a crucial component for effective cellulose decomposition.

Usage and Biology

Kluyveromyces marxianus mating factor alpha is a secretion signal in Kluyveromyces marxianus. The MFa encodes for an alpha mating factor domain that is fused with TrEGIII-t. The translation of this signal peptide found on the MFa domain causes the polypeptide to enter RER and Golgi body and allows the target enzyme to be secreted by Kluyveromyces marxianus.

TrEGIII-t is type of endo-1,4-beta-endoglucanase with type I dockerin fused to its C terminal. The enzyme functions by hydrolyzing internal glycosidic bonds in cellulose chains to generate new free ends that CBH can acts on. TrEGIII-t, CBHII-t, and NpaBGS-t works collectively with the assistance of two cellulase boosters to achieve a highly-efficient degradation of cellulose in waste textiles.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 331
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 331
  Illegal NheI site found at 89
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 331
  Illegal XhoI site found at 1252
  Illegal XhoI site found at 2830
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 331
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 331
  Illegal AgeI site found at 1603
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1475