CELLULOLYTIC POLYPEPTIDES AND THEIR USE IN MICRO-ORGANISMS FOR THE PRODUCTION OF SOLVENTS AND FUELS
The invention relates to applications of a cellulase of Pseudomonas sp. ND137, functional fragments and/or variants and engineered forms thereof, in the context of industrial bioprocessing, more particularly solvent production.
1 . A method for producing a solvent, fuel or chemical intermediate from a substance comprising cellulose, comprising:
a) expressing the ACLA polypeptide of Pseudomonas sp. ND137 or a homologue thereof, or a functional fragment and/or variant of said ACLA polypeptide or of said homologue in a recombinant host cell, and
b) contacting said substance with said ACLA polypeptide.
2 . The method of claim 1 , which further comprises the step of separating said ACLA enzyme or active fragment from said recombinant host cell.
3 . A recombinant micro-organism expressing the ACLA polypeptide of Pseudomonas sp. ND137 or a homologue thereof, or a functional fragment and/or variant of said ACLA polypeptide or of said homologue, wherein said micro-organism is capable of degrading cellulose.
4 . The recombinant micro-organism according to claim 3 which is a solventogenic micro-organism.
5 . The recombinant solventogenic micro-organism according to claim 4 , which is an ethanologenic micro-organism.
6 . The recombinant solventogenic micro-organism according to claim 4 which is a solventogenic bacterium.
7 . The recombinant solventogenic bacterium according to claim 6 , which is Clostridium acetobutylicum.
8 . The method according to claim 1 , comprising contacting said substance with a recombinant micro-organism expressing the ACLA polypeptide of Pseudomonas sp. ND137 or a homologue thereof, or a functional fragment and/or variant of said ACLA polypeptide or of said homologue, wherein said micro-organism is capable of degrading cellulose.
9 . A method for the degradation of a substance comprising cellulose, comprising contacting said substance with the recombinant micro-organism according to claim 3 .
10 . The method according to claim 1 , wherein said substance comprises or is enriched in crystalline cellulose.
11 . A recombinant micro-organism expressing
(a) the ACLA polypeptide of Pseudomonas sp. ND137 or a homologue thereof, or a functional fragment and/or variant of said ACLA polypeptide or of said homologue, and
(b) one or more additional foreign cellulose-degrading enzymes other than said ACLA polypeptide.
12 . A mixture of enzymes comprising the ACLA polypeptide of Pseudomonas sp. ND137 or a homologue thereof, or a functional fragment and/or variant of said ACLA polypeptide or of said homologue and one or more cellulose-degrading enzymes other than said ACLA polypeptide.
13 . A method for the production of a solvent, fuel, or chemical intermediate from a substance comprising cellulose, wherein said method comprises contacting said substance with
the ACLA polypeptide of Pseudomonas sp. ND137 or a homologue thereof, or a functional fragment and/or variant of said ACLA polypeptide or of said homologue.
14 . The method according to claim 8 , wherein the micro-organism is a solventogenic micro-organism.
15 . The method according to claim 14 , wherein the solventogenic microorganism is a solventogenic bacterium.
16 . The method according to claim 15 , wherein the solventogenic bacterium is Clostridium acetobutylicum.
17 . A method for the degradation of a substance comprising cellulose, comprising contacting said substance with the recombinant micro-organism according to claim 4 .
18 . A method for the degradation of a substance comprising cellulose, comprising contacting said substance with the recombinant micro-organism according to claim 5 .
19 . A method for the degradation of a substance comprising cellulose, comprising contacting said substance with the recombinant micro-organism according to claim 6 .
20 . A method for the degradation of a substance comprising cellulose, comprising contacting said substance with the recombinant micro-organism according to claim 7 .