Part:BBa_K3803016

This is a efficient xylose utilization composite part.

Based on the parts (BBa_K2314913 & BBa_K2314324) from OUC_China in 2017 iGEM, we built a more efficient xylose utilization part BBa_K3803016. Xylose reductase (XR) will first transform xylose to xylitol, and xylitol dehydrogenase (XDH) can further convert xylitol to xylulose.Then, xylulose will be converted to xylulose 5-phosphate (X5P) by the native xylulose kinase (XK). Apart from the original XR and XDH genes, considering the low copy number of native XK gene in S. cerevisiae, we introduced an extra XK gene to improve the xylose utilization ability. You can see the detailed improvement here:

For cell growth, OUC_China achieved a final OD600 around 2.35 in YNB-based xylose media. After our improvement on their parts, we could get a final OD600 around 20. You can see the comparison of OD after improvement here:

For our own product yield, our production also improved after substituting our three-gene cassette for OUC_China's two-gene one. You can see the HPLC results with different xylose utilization genes here:

For more information about our parts and improvement, you can click here: https://2021.igem.org/Team:Jiangnan_China/Parts

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 805
  Illegal XbaI site found at 6464
  Illegal PstI site found at 2114
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 805
  Illegal PstI site found at 2114
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 805
  Illegal BglII site found at 1265
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 805
  Illegal XbaI site found at 6464
  Illegal PstI site found at 2114
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 805
  Illegal XbaI site found at 6464
  Illegal PstI site found at 2114
  Illegal AgeI site found at 1936
  
• 1000
  COMPATIBLE WITH RFC[1000]