Method to produce alcohol and alcohol precursors using genetically modified bacteria

A method to obtain an alcohol or alcohol precursor from a lignocellulosic biomass and a genetically modified live ethanologenic organism, wherein said process comprising the steps of:

• providing a lignocellulosic biomass;
• exposing said lignocellulosic biomass to a modified Caro's acid composition for a period of time necessary to remove more than 98.5% of the lignin present in said lignocellulosic biomass and thus obtaining a source of cellulose and a liquid stream;
• separating said solid stream comprising said source of cellulose from said liquid stream comprising said lignin;
• exposing said source of cellulose to said genetically modified live ethanologenic organism in a culture media with a pH of between 2 and 9, thereby creating an incubation mixture;
• incubating said genetically modified live ethanologenic organism in said incubation mixture with said source of cellulose under aerobic and/or anaerobic conditions at a temperature ranging from 0° C. to 60° C. for a first period of time sufficient for the degradation of cellulose, thereby creating a degraded cellulose mixture;
• optionally, incubating said genetically modified live ethanologenic organism in said degraded cellulose mixture under aerobic and/or anaerobic conditions at a temperature ranging from 0° C. to 60° C. for a second period of time sufficient for the production of said alcohol or alcohol precursor;
• and
• optionally, recovering said alcohol or alcohol precursor from the cells and/or spent culture media;
• wherein said genetically modified live ethanologenic organism comprises at least one of the following polynucleotide sequences in its genome:
• i. at least one endoglucanase (cen-like) polynucleotide sequence selected from the group consisting of an endoglucanase A-like (cenA-like) polynucleotide sequence, an endoglucanase B-like (cenB-like) polynucleotide sequence and an endoglucanase C-like (cenC-like) polynucleotide sequence;
• ii. an exoglucanase (cex-like) polynucleotide sequence;
• and
• iii. a β-glucosidase 1 (bgl1) polynucleotide sequence;
• wherein said live ethanologenic organism belongs to a genus is selected from the group consisting of:
• Aspergillus, Mucor, Zymomonas, Escherichia, Clostridia, Bacillus, and Pseudomonas.

A method of growing a genetically modified live ethanologenic organism, wherein said method comprises the steps of:

• exposing said live ethanologenic organism to a culture media with a pH of between 2 and 9 comprising a carbon source and a nitrogen source, thereby creating an incubation mixture;
• and
• incubating said live ethanologenic organism in said incubation mixture under aerobic and/or anaerobic conditions at a temperature ranging from 0° C. to 60° C. for a period of time sufficient to allow for the exponential growth of said live ethanologenic organism.
• and wherein said live ethanologenic organism comprises at least one of the following polynucleotide sequences in its genome:
• i. an endoglucanase (cen-like) polynucleotide sequence selected from the group consisting of an endoglucanase A-like (cenA-like) polynucleotide sequence, an endoglucanase B-like (cenB-like) polynucleotide sequence and an endoglucanase C-like (cenC-like) polynucleotide sequence;
• ii. an exoglucanase (cex-like) polynucleotide sequence;
• and
• iii. a β-glucosidase 1 (bgl1) polynucleotide sequence.
• and wherein said live ethanologenic organism belongs to a genus is selected from the group consisting of:
• Aspergillus, Mucor, Zymomonas, Escherichia, Clostridia, Bacillus, and Pseudomonas.