Part:BBa_K5159004

Schizochytrium sp. ATCC 20888 PPTase

Phosphopantetheinyl transferase (PPTase) is required for the proper function of PUFA synthase. It activates the acyl-carrier protein (ACP) domains on the PUFA synthase subunit A (Part:BBa_K5159001) so that they can bind to their substrates under help of the malonyl-ACP transferase (MAT) domain. Therefore in our project, it is co-expressed with the 3 PUFA synthase ORFs in oleaginous yeast chassis Yarrowia lipolytica. The sequence 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.

This PPTase is specific to the PUFA synthase utilized in our project for DHA production, while the absence of the PPTase is reported to completely disable the DHA synthesis process (Hauvermale et al, 2006). Compared to PPTase identified in prokaryotic organisms such as Moritella marina, this PPTase originated from Schizochytrium sp. shows an increased catalytic efficiency (Wang et al, 2020).

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 322
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 19
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 121
  Illegal SapI.rc site found at 736
  

Reference

[1] Hauvermale, A., Kuner, J., Rosenzweig, B. et al (2006). Fatty acid production in Schizochytrium sp.: Involvement of a polyunsaturated fatty acid synthase and a type I fatty acid synthase. Lipids 41, 739–747. https://doi.org/10.1007/s11745-006-5025-6

[2] Wang, S., Lan, C., Wang, Z. et al (2020). PUFA-synthase-specific PPTase enhanced the polyunsaturated fatty acid biosynthesis via the polyketide synthase pathway in Aurantiochytrium. Biotechnol Biofuels 13, 152. https://doi.org/10.1186/s13068-020-01793-x