Design
We design two parts.[preventation & elimination]
We design two parts.[preventation & elimination]
1) Reaction: NO2- + 2H+ + 2e- -> NO + H2O
2) 3D-view: 1NIR
1) Reaction: NO + NH4+ 2H+ + 2e- -> N2H4 + H2O
2) 3D-view: 5C2V / Red: Alpha, Blue: Beta, Yellow: Gamma
1) Reaction: NO + NH4+ 2H+ + 2e- -> N2H4 + H2O
2) 3D-view: 4N4L
1) Reaction: 2O2- + 2H+ -> H2O2 + O2
2) 3D-view: 2JLP
1. What is Cell Free Expression System
It is a system that facilitates in vitro synthesis of target proteins by reacting whole cell extracts and nuclear extracts, or genetic material such as DNA or RNA, outside living cells. As no other cellular activity – except for transcription and translation – occurs, protein synthesis is accelerated, resulting higher yield and efficiency. Compared to conventional in vivo protein expression systems, CFE is much less labourious due to its reduced and simplified experimental procedure.
2. Problem facing current Anammox process
Anammox process is one of the most efficient ways of treating wastewater, particularly nitrogen-based nutrients. However, there is a critical problem with the current process using living Anammox bacteria, arisen from the use of “living cell”. Anammox bacteria require a strict growth condition and their growth rate is extremely low to the extent of over 2 months of complete growth, which implies that the culturing of those bacteria is a challenging and costly task. This serves as a barrier in introducing Anammox process, a highly efficient and effective technique, extensively.
3. Application of CFE to Anammox process
We came up with the idea of grafting CFE to the Anammox process as a solution to the above-stated problem. CFE facilitates a mass production of nitrogen removal enzymes without the need of culturing bacteria. Enzymes produced from CFE can be attached to porous membrane and put into wastewater needed to be treated. We believe our solution can mimic anammox process with much shorter time and less effort, and contribute to the effective removal of nitrogen from any wastewater abundant with nitrogen nutrients.
This part is Preventation part which BBa_K4371000 (Anammox cluster: HZS + HZO + NIR) is inserted into the pEXP5-NT(expression vector).
pEXP5-NT is specific cell free expression(CFE) vector.
This part is Elimination part which BBa_K38007 (SOD1 protein) is inserted into the pEXP5-NT(expression vector).
pEXP5-NT is specific cell free expression(CFE) vector.
