Part:BBa_K5159008

NADP+-dependent GapC gene from Clostridium acetobutylicum

The GapC gene encodes a NADP+-dependent G3P dehydrogenase when introduced into oleaginous yeast Yarrowia lipolytica, which converts NADP+ to NADPH, increasing its availability during fatty acid (e.g., DHA) synthesis. This is achieved by the enzyme's substrate selectivity enabling cell metabolism to prefer using NADP+ instead of normally NAD+ during glycolysis to provide the oxidation power. It can also potentially enhance generation of acetyl-CoA, the main substrate for fatty acid synthesis, which locates downstream of the introduced metabolic pathway.

This sequence from Clostridium acetobutylicum EA 2018 is codon-optimized for Yarrowia lipolytica and a His-tag with a linker sequence is attached to the C terminus to facilitate the enzyme expression detection.

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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 46
  

Reference

[1] Qiao, K., Wasylenko, T., Zhou, K. et al (2017). Lipid production in Yarrowia lipolytica is maximized by engineering cytosolic redox metabolism. Nat Biotechnol 35, 173–177. https://doi.org/10.1038/nbt.3763

[2] Glyceraldehyde 3-phosphate dehydrogenase, gene gapC [ Clostridium acetobutylicum EA 2018 ], Gene ID: 12232727. https://www.ncbi.nlm.nih.gov/gene/12232727