Part:BBa_K2833022

J23104+B0032+VHb+Ter

Vitreoscilla haemoglobin (VHb) is a type of haemoglobin found in the Gram-negative aerobic bacterium, Vitreoscilla. It is the first haemoglobin discovered from bacteria, but unlike classic hemoglobin it is composed only of a single globin molecule.

Previous research has found that expressing Vitreoscilla hemoglobin (VHb) inside the bacteria could leverage its property of combining and releasing oxygen to improve the efficiency of oxygen utilization and therefore enhance the efficiency of bacteria growth and production.^[1]

This year our project focused on enhancing the growth and production efficiency of the engineered bacteria during high-density fermentation, to achieve this we linked the VHb gene(got from Chen Guoqiang Lab in Tsinghua University) to the downstream of J23104 and B0032 to construct this part(figure 1) and compared the growth curve of BBa_K2833022 transformed bacteria and empty vector transformed bacteria to see if the VHb works(figure 2).

iGEM14_Imperial has constructed BBa_K1321200 which can express VHb under the control of J23101 and B0032. However, when making alignment we found that the sequence of VHb in BBa_K1321200 was different from the original VHb both in DNA and protein.(figure 3). So we also compared the growth curve with BBa_K1321200 transformed bacteria to see which one works better.(figure 2)

From the result we can see that both the two VHb transgene bacteria grew better than the wildtype bacteria, mainly presented on the maximal bio-mass at platform, while our newly built constructs BBa_K2833022 showed bigger effects on the growth of the bacteria.

Figure 1. The framework of BBa_K2833022

Figure 2. The growth curve of wildtype and VHb transgene bacteria

Figure 3. Alignment for the VHb sequence in BBa_K1321200 with the VHb sequence in BBa_K2833022(as found in NCBI).A: alignment of the DNA sequence; B: Alignment of the protein sequence.

References

1.Lei Zhang et al.. Recent developments and future prospects of Vitreoscilla hemoglobin application in metabolic engineering. Biotechnology Advances 25(2007).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]