Part:BBa_K5314004

AM. Sed + VHb

Vitreoscilla hemoglobin (VHb) is a type of bacterial hemoglobin from Vitreoscilla. It has been expressed in a variety of organisms [1] and is a widely used synthetic biological hemoglobin. It can be highly expressed at low oxygen concentrations. Its function may be to transport oxygen to oxidases at the end of the electron transport chain, thereby increasing the efficiency of oxygen uptake. [2] It is suitable for anoxic environments such as high-density cell fermentation. [3]

Our project focuses on further improving the efficiency of VHb protein expression for use in our high-density fermentation production. We decide to add cell wall anchoring proteins [4] to secrete VHb into the cell wall to receive oxygen and use the concentration difference to transport oxygen into the cell.

We use cell wall anchoring sequence, AM.Sed from Aureobasidium Melanogenum BZ-11, and each is connected to the wild-type VHb.

Fig 1. The 3D structure of AM.Sed VHb (Predicted by AlphaFold 3)

Source

Organism: VHb (Vitreoscilla stercoraria), AM.Sed (Aureobasidium Melanogenum BZ-11)

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 762
  
• 1000
  COMPATIBLE WITH RFC[1000]

References:
[1] Khosla, C., & Bailey, J. E. (1988). Heterologous expression of a bacterial haemoglobin improves the growth properties of recombinant Escherichia coli. Nature, 331(6157), 633–635. https://doi.org/10.1038/331633a0

[2] Ramandeep, Hwang, K. W., Raje, M., Kim, K. J., Stark, B. C., Dikshit, K. L., & Webster, D. A. (2001). Vitreoscilla hemoglobin. Intracellular localization and binding to membranes. The Journal of biological chemistry, 276(27), 24781–24789. https://doi.org/10.1074/jbc.M009808200

[3] Zhang, L., Li, Y., Wang, Z., Xia, Y., Chen, W., & Tang, K. (2007b). Recent developments and future prospects of Vitreoscilla hemoglobin application in metabolic engineering. Biotechnology Advances, 25(2), 123–136. https://doi.org/10.1016/j.biotechadv.2006.11.001

[4] Jaafar, L., & Zueco, J. (2004). Characterization of a glycosylphosphatidylinositol-bound cell-wall protein (GPI-CWP) in Yarrowia lipolytica. Microbiology (Reading, England), 150(Pt 1), 53–60. https://doi.org/10.1099/mic.0.26430-0