Team:UAAAN/Basic Part
Characterization of the cluster of H + producing genes, obtained from Ruminococcus albus
Considering the information generated with Clamydomonas rehindardti, the methodology for the characterization of a new BioBrick was proposed, this time taking the sequences of the bacterium Ruminococcus albus. R. albus is among the most important bacteria that degrade the wall of plant cells in the rumen. These species produce all the enzymes required for the hydrolysis of plant cell wall polysaccharides, cellulose and hemicellulose (Zheng et al., 2014). Its most important characteristics are:
°°Morphology:
Gram: Positive
Shape: Cocci
Motility: No
°°Environment:
Oxygen Requirements: Anaerobic
Temperature range: Mesophilic
Habitat: Associated with the guest
On this occasion the cluster of genes involved in the production of Hydrogen in this bacterium was also considered (Zheng et al., 2014). Which is described and schematized below:
FIG1. Scheme of the energy metabolism of R. albus 7 growing on glucose at low (a = 0) and at very high (a = 1) H2 partial pressures. The relevant reaction is glucose + (2-a) H2O-> 2a ethanol + 2CO2 + (2-a) acetate- + (2-a) H + + (4-2a) H2
HydABC (highlighted in red) is electron-dependent ferredoxin and NAD-dependent [FeFe] -hydrogenase, and HydA2 (green highlighted) is ferredoxin-dependent [FeFe] -hydrogenase. The presence of the three membrane protein complexes is deduced from the genome sequence of said bacterium Ruminococcus albus (Zheng et al., 2014).
The sequences of the HydA, HydB and HydC genes are found in the chromosome of the bacteria and form the final maturation enzyme complex in the H2 production process.
FIG2.Hypothetical location of the HydABC complex within the Ruminococcus albus chromosome
Basic BioBricks BBa_K2998001 and BBa_K2998004
On the other hand, the HydA2 and HydS protein complexes that make up our cluster of genes of interest are found in one of the 4 plasmids that the bacteria have
Ruminococcus albus sequences were obtained from NCBI (Suen et al., 2011).
REFERENCES
Suen, G., Stevenson, D. M., Bruce, D. C., Chertkov, O., Copeland, A., Cheng, J. F., … Weimer, P. J. (2011). Complete genome of the cellulolytic ruminal bacterium Ruminococcus albus 7. Journal of Bacteriology, 193(19), 5574–5575. https://doi.org/10.1128/JB.05621-11
Zheng, Y., Kahnt, J., Kwon, I. H., Mackie, R. I., & Thauer, R. K. (2014). Hydrogen formation and its regulation in Ruminococcus albus: Involvement of an electron-bifurcating [FeFe]-hydrogenase, of a non-electron-bifurcating [FeFe]-hydrogenase, and of a putative hydrogen-sensing [FeFe]-hydrogenase. Journal of Bacteriology, 196(22), 3840–3852. https://doi.org/10.1128/JB.02070-14