# US 4,400,470 A · Use of co-cultures in the production of ethanol by the fermentation of biomass

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- Number: US 4,400,470 A
- Document ID: US4400470A
- Status: Granted
- Kind: A
- Application number: 06/225,093
- Filed: 1981-01-14
- Granted: 1983-08-23
- Earliest claimed: 1981-01-14
- Assignees: Wisconsin Alumni Research Foundation
- Inventors: Joseph G. Zeikus; Thomas K. Ng; Arie Ben-Bassat; Raphael J. Lamed
- Web page: https://patentgrep.com/patent/US4400470A

## CPC

- C12N 9/0006: {acting on CH-OH groups as donors (1.1)}
- C12N 9/2437: {Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)}
- C12P 7/08: produced as by-product or from waste or cellulosic material substrate
- C12P 39/00: Processes involving microorganisms of different genera in the same process, simultaneously
- C12Y 302/01004: Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
- Y02E 50/10: Biofuels, e.g. bio-diesel
- Y10S 435/842: Clostridium
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES
- Y02E: REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

Production of ethanol and enzymes by fermentation of biomass with co-cultures of C. thermocellum and C. thermohydrosulfuricum.

## Claims

### Claim 1

In the method of producing ethanol and enzymes as a by-product of ethanol production by the fermentation of biomass, the improvement comprising subjecting the biomass which includes cellulose and hemi-cellulose containing plant materials to fermentation under thermophilic conditions in the presence of co-cultures of Clostridium thermocellum and Clostridium thermohydrosulfuricum, the ratio of the cultures being in an amount whereby the conversion ratios of cellulose:ethanol and hemi-cellulose:ethanol are greater than the ratios obtained by use of either Clostridium species alone.

### Claim 2

In the method of producing ethanol and enzymes as a by-product of ethanol production by the fermentation of biomass, the improvement comprising subjecting the biomass which includes cellulose and hemi-cellulose containing plant materials to fermentation by Clostridium thermohydrosulfuricum under thermophilic conditions in the presence of the elaborated enzymes produced by Clostridium thermocellum, the ratio of the Clostridium thermohydrosulfuricum to the elaborated enzymes being in an amount whereby the conversion ratios of cellulose:ethanol and hemi-cellulose:ethanol are greater than the ratios obtained by use of either Clostridium species alone.

### Claim 3 (depends on 1)

The method as claimed in claim 1 in which the fermentation is carried out at a temperature of about 60.degree.-65.degree. C.

### Claim 4 (depends on 1, 2)

The method as claimed in claims 1 or 2 in which the enzymes produced comprise extra cellular cellulase, cell bound reversible NADP linked alcohol dehydrogenase and cell bound malic enzyme.

### Claim 5 (depends on 1, 2)

The method as claimed in claims 1 or 2 in which the fermentation is carried out anaerobically in the presence of hydrogen maintained at a pressure sufficient to promote ethanol formation but inhibit hydrogen and acetate formation.