Process for the production of an enzymatic cocktail using liquid residues from a process for the biochemical conversion of lignocellulosic materials

• US 10,030,236 B2
• Filed: 06/10/2013
• Issued: 07/24/2018
• Est. Priority Date: 06/18/2012
• Status: Active Grant

• Current Assignee: Agro industrie Recherches et Développements, Institut national de recherche pour l'agriculture, l'alimentation et l'environnement
• Original Assignee: Agro industrie Recherches et Développements, Institut National de La Recherche Agronomique
• Inventors: Ben Chaabane, Fadhel, Louret, Sylvain
• Primary Examiner(s): Fan, Lynn Y
• Application Number:
  US14/408,612
• Publication Number:
  US 20150152400A1
• Time in Patent Office: 1,870 Days
• CPC Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12N 9/2437 Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
• CPC Class Codes: C12N 9/2445 Beta-glucosidase (3.2.1.21)
• CPC Class Codes: C12P 21/00 Preparation of peptides or proteins single cell protein C12N1/00
• CPC Class Codes: C12P 2203/00 Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source ethanol C12P7/10
• CPC Class Codes: C12Y 302/01004 Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel

Abstract

A process for the production of an enzymatic cocktail with a cellulolytic microorganism comprises two phases:

• • a phase a) for growth of said microorganism in a closed reactor in the presence of a carbonaceous growth solution;
  • a phase b) for the production of said enzymatic cocktail carried out with a supply of carbonaceous production solution the concentration of carbonaceous substrate of which is in the range 150 to 400 g/L, said carbonaceous production solution comprising a carbonaceous inducer substrate;
    characterized in that said carbonaceous inducer substrate is a liquid residue obtained from a step for pre-treatment of lignocellulosic materials, the C₅ sugar oligomers of which represent at least 1% by weight of the total sugars present in said liquid residue, and at least 0.3% by weight of the total sugars present in said carbonaceous production solution.

Claims

1. A process for the production of an enzymatic cocktail with a cellulolytic microorganism, comprising:

pre-treating lignocellulosic materials without sterilization or modification of the pH of a liquid residue obtained therefrom,

growing a microorganism in a closed reactor in the presence of a carbonaceous growth solution comprising a carbonaceous growth substrate;

producing an enzymatic cocktail comprising a supply of carbonaceous production solution including a concentration of a carbonaceous substrate in the range 150 to 400 g/L, wherein the carbonaceous production solution comprises a carbonaceous inducer substrate, wherein the carbonaceous inducer substrate is the obtained liquid residue and comprises C₅ sugar oligomers,

wherein the C₅ sugar oligomers are at least 1% by weight of the total sugars present in said liquid residue, and are 0.3% to 20% by weight of the total sugars present in said carbonaceous production solution wherein the C₅ sugar oligomers in the carbonaceous inducer substrate induce the production of enzymes.

2. The process of claim 1, wherein in the growing step the carbonaceous growth solution is present in an initial concentration of 10 to 90 g of carbonaceous substrate per liter of reaction volume.

3. The process of claim 1, wherein said pre-treatment step is acid hydrolysis, acid cooking, or steam explosion with prior impregnation of said lignocellulosic materials with an aqueous sulphuric acid solution.

4. The process of claim 1, wherein said liquid residue is not detoxified.

5. The process of claim 1, wherein the C₅ sugar oligomers are 1% to 50% by weight of the total sugars present in said liquid residue.

6. The process of claim 1, wherein said carbonaceous inducer substrate is used alone or as a mixture with at least one non-inducer carbonaceous substrate.

7. The process of claim 6, wherein said non-inducer carbonaceous substrate is glucose, xylose or saccharose, alone or as a mixture.

8. The process of claim 1, wherein said carbonaceous production solution consists of the carbonaceous inducer substrate and at least one non-inducer carbonaceous substrate which is glucose, xylose or saccharose, alone or as a mixture,

wherein said liquid residue is obtained without detoxification of said liquid residue, and consists of C₅ sugar oligomers, C₅ and C₆ sugar monomers, and sugar monomer degradation products.

9. The process of claim 1, wherein a specific flow rate for supplying carbonaceous production solution is in the range 35 to 65 mg of carbonaceous inducer substrate per gram of microorganism per hour.

10. The process of claim 1, wherein the cellulolytic microorganism is strains of Trichoderma, Aspergillus, Penicillium or Schizophyllum.

11. The process of claim 10, wherein the cellulolytic microorganism is strains of Trichoderma reesei.