BIOCATALYSTS SYNTHESIZING DEREGULATED CELLULASES
Provided are isolated novel Clostridium phytofermentans biocatalysts with deregulated cellulase activity that produce high yields of products. Further provided are methods of using the biocatalysts to degrade organic material and for use in industrial processes.
1 . An isolated microorganism that produces a fermentation end-product from a biomass, said microorganism comprising a genetic modification that enables said microorganism to synthesize more cellulases in the presence of an inhibitor molecule than a microorganism of the same species without said genetic modification.
2 . The microorganism of claim 1 , wherein said inhibitor molecule is glucose or a glucose analog.
3 . The microorganism of claim 1 , wherein said genetic modification comprises a mutation in one or more genes, wherein at least one of said genes encodes a propionyl-CoA carboxylase, a two component AraC family transcriptional regulator, or a ROK family glucokinase, a homolog of Cphy — 3487, a dihydrolipoamide dehydrogenase, a binding-protein-dependent transport systems inner membrane component, an ABC transporter related protein, a homolog of Cphy — 0056, a TetR family transcriptional regulator, an AraC-like protein, a glycosyl transferase 36, a diaminopimelate epimerase, an oxidoreductase domain-containing protein, a homolog of Cphy — 2965, a desulfoferrodoxin ferrous iron-binding region, a homolog of Cphy — 1063, an AraC family transcriptional regulator, a phage tape measure protein, a D-isomer specific 2-hydroxyacid dehydrogenase NAD-binding protein, or an HD superfamily phosphohydrolase-like protein.
4 . The microorganism of claim 1 , wherein said fermentation end-product is an alcohol.
5 . The microorganism of claim 4 , wherein said alcohol is ethanol.
6 . The microorganism of claim 1 , wherein said biomass comprises hemicellulosic or lignocellulosic material.
7 . The microorganism of claim 1 , wherein said microorganism can hydrolyze and ferment hemicellulosic or lignocellulosic material.
8 . The microorganism of claim 1 , wherein said microorganism is a Clostridium species.
9 . A method of producing a fermentation-end product comprising:
a. providing a biomass in a media;
b. contacting said biomass with an isolated microorganism comprising a genetic modification that enables said microorganism to synthesize more cellulases in the presence of an inhibitor molecule than a microorganism of the same species without said genetic modification; and,
c. allowing sufficient time for said microorganism to produce said fermentation end-product from said biomass.
10 . The method of claim 9 , wherein said inhibitor molecule is glucose or a glucose analog.
11 . The method of claim 9 , wherein said fermentation end-product is an alcohol.
12 . The method of claim 11 , wherein said alcohol is ethanol.
13 . The method of claim 9 , wherein said biomass comprises hemicellulosic or lignocellulosic material.
14 . The method of claim 9 , wherein said microorganism can hydrolyze and ferment hemicellulosic or lignocellulosic material.
15 . The method of claim 9 , wherein said microorganism is a Clostridium species.
16 . A plant for producing a fermentation end product comprising:
a. a fermenter, wherein said fermenter is configured to house a biomass in a medium; and,
b. an isolated microorganism comprising a genetic modification that enables said microorganism to synthesize more cellulases in the presence of an inhibitor molecule than a microorganism of the same species without said genetic modification.
17 . The plant of claim 16 , wherein said inhibitor molecule is glucose or a glucose analog.
18 . The plant of claim 16 , wherein said fermentation end-product is an alcohol.
19 . The plant of claim 18 , wherein said alcohol is ethanol.
20 . The plant of claim 16 , wherein said biomass comprises hemicellulosic or lignocellulosic material and wherein said microorganism can hydrolyze and ferment said hemicellulosic or lignocellulosic material.