Design
Design: Gelectricell
Our EMFC has 3 main systems.
1.Agar degradation
We used 3 enzymes to degrade agar in our device which are, Agarase, NABH (Neoagarobiose hydrolase), and AHGD (anhydrogalactose dehydrogenase).
Agar is first degraded into neoagarobiose by agarase. Neoagarosebiose is then degraded into D-galactose and 3.6-anhydro-L-galactose by NABH. 3.6-anhydro-L-galactose is then oxidized by NAD(P)+ dependent AHGD producing NAD(P)H.
All listed enzymes are displayed on the surface of E.coli BW25113 using E.coli surface display vector pATLIC.
2. Shewanella oneidensis MR-1
Shewanella oneidensis MR-1 is a bacteria that can reduce metal instead of oxygen, thus generating electricity in a battery device.
Shewanella oneidesis MR-1 is used in our device to generate electricity using D-galactose. However, Shewanella oneidensis MR-1 is known to be unable to use galactose as its carbon source. This is where co-cultured E.coli kicks in. E.coli BW25113 is able to utilize galactose to produce formate and acetate which Shewanella oneidensis MR-1 can utilize to generate electricity.
3. Diaphorase
Diaphorase is a type of enzyme that can generate electricity using NAD(P)H as the source of electron. It is expressed in BL21(DE3) with protein expression vector pB3.
References
- Yun, Eun Ju, et al. "Production of 3, 6-anhydro-L-galactose from agarose by agarolytic enzymes of Saccharophagus degradans 2-40." Process biochemistry 46.1 (2011): 88-93.
- Yun, Eun Ju, et al. "The novel catabolic pathway of 3, 6‐anhydro‐L‐galactose, the main component of red macroalgae, in a marine bacterium." Environmental microbiology 17.5 (2015): 1677-1688.
- Ko, Hyeok-Jin, et al. "Functional cell surface display and controlled secretion of diverse agarolytic enzymes by Escherichia coli with a novel ligation-independent cloning vector based on the autotransporter YfaL." Applied and environmental microbiology 78.9 (2012): 3051-3058.
- Wang, Victor Bochuan, et al. "Metabolite-enabled mutualistic interaction between Shewanella oneidensis and Escherichia coli in a co-culture using an electrode as electron acceptor." Scientific reports 5 (2015).