Lignocellulosic biomass is generally regarded as a sustainable and environmentally friendly energy source.
However, the production of fuels from biomass is not sufficiently effective to compete fully with fossil fuels on economic terms, mostly due to the low efficiency of the enzymes needed to degrade the biomass.
Lignocellulose is the most abundant biomass on earth and has a great potential as a source for sustainable production of valuable chemicals and biofuels.
Today’s depletion of fossil fuel reserves and pollution from their usage creates a need for a more sustainable source of energy.
The cluster enzyme characterization gives insight into the use of the Type IX secreted multi-domain cellulases.
Further work on the gene cluster can provide more insight into the degradation mechanism, and expression of the cluster in a bacterium harbouring the Type IX secretion system could be pursued to improve the expression of the multi-domain cellulases that were difficult to express.
The aim of the present study was to clone, express and purify these GH5 enzymes for biochemical characterization to gain insight into their substrate targets and functional roles in the cluster.
The enzymes, together with truncated versions containing only one domain, were cloned, expressed and purified.
Denne oppgaven ble basert på en metagenomisk studie av en switchgrass prøve fra kumage, hvor videre undersøkelser fant et antatt cellulolytisk genkluster, fra en Bacteroidetes fylotypes genom (AGa), med fire antatt cellulolytiske glykosid-hydrolase familie 5 (GH5) enzymer.
Klusteret inneholdt både multi-domene enzymer og enzymer med et C-terminalt sekresjonssignal for Type IX sekresjonssystemet (T9SS).