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Single-step bioconversion of lignocellulosic biomass to biofuels using extreme thermophilic bacteria

a technology of lignocellulosic biomass and biofuel, which is applied in the field of single-step bioconversion of lignocellulosic biomass to biofuels using extreme thermophilic bacteria, can solve the problems of increasing food costs, facing the challenge of redirecting the production process, and cellulosic biomass is a vast poorly exploited resource, and achieves high levels of biofuels

Inactive Publication Date: 2014-12-11
DIREVO INDAL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One challenge associated with this strategy is that competition between food markets and energy markets for the crops can increase food costs.
However, the industry of producing fermentation products such as ethanol is facing the challenge of redirecting the production process from fermentation of relatively easily convertible but expensive starchy materials, to the complex but inexpensive lignocellulosic biomass such as plant biomass.
Cellulosic biomass is a vast poorly exploited resource, and in some cases a waste problem.
Each processing step can make the overall process more costly and, therefore, decrease the economic feasibility of producing biofuel or carbon-based chemicals from cellulosic biological material.

Method used

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  • Single-step bioconversion of lignocellulosic biomass to biofuels using extreme thermophilic bacteria
  • Single-step bioconversion of lignocellulosic biomass to biofuels using extreme thermophilic bacteria
  • Single-step bioconversion of lignocellulosic biomass to biofuels using extreme thermophilic bacteria

Examples

Experimental program
Comparison scheme
Effect test

example 1

Isolation and Cultivation

[0140]All procedures for enrichment and isolation of the strains listed in table 1 employed anaerobic technique for strictly anaerobic bacteria (Hungate 1969). The strains were enriched from environmental samples at temperatures higher than 70° C. with crystalline cellulose and beech wood as substrate. Isolation was performed by picking colonies grown on solid agar medium at 72° C. in Hungate roll tubes (Hungate 1969).

[0141]The cells are cultured under strictly anaerobic conditions applying the following medium:

Basic mediumNH4Cl1.0gNaCl0.5gMgSO4 × 7 H2O0.3gCaCl2 × 2 H2O0.05gNaHCO30.5gK2HPO41.5gKH2PO43.0gYeast extract (bacto, BD)0.5gCellobiose5.0gVitamins (see below)1.0mlTrace elements (see below)0.5mlResazurin1.0mgNa2S × 9 H2O0.75gDistilled water1000.0mlTrace elements stock solutionNiCl2 × 6H2O2gFeSO4 × 7H2O1gNH4Fe(III) citrate, brown, 21.5% Fe10gMnSO4 × H2O5gCoCl2 × 6H2O1gZnSO4 × 7H2O1gCuSO4 × 5H2O0.1gH3BO30.1gNa2MoO4 × 2H2O0.1gNa2SeO3 × 5H2O0.2gNa2WoO4 × 2...

example 2

HPLC

[0145]Sugars and fermentation products were quantified by HPLC-RI using a Via Hitachi LaChrom Elite (Hitachi corp.) fitted with an Rezex ROA Organic Acid H+ (Phenomenex). The analytes were separated isocratically with 2.5 mM H2SO4 and at 65° C.

example 3

Phylogenetic Analysis of 16S rDNA Genes

[0146]Genomic DNA was isolated from cultures grown as described above and 16SrDNA amplified by PCR using 27F (AGAGTTTGATCMTGGCTCAG; SEQ ID No. 8) as forward and 1492R (GGTTACCTTGTTACGACTT; SEQ ID No. 9) as reverse primer. The resulting products were sequenced and the sequences analyzed using the Sequencher 4.10.1 software (Gene Codes Corporation). The NCBI database was used for BLAST procedures.

[0147]Sequencing of 16S rDNA from all strains listed in table 1 revealed all these had (at least) one copy of a 16S rDNA operon which was most closely related to Caldicellulosiruptor saccharolyticus (Strain Tp8T=DSM8903) in the available public databases. Alignment was carried out using ClustalW (Chenna et al. 2003) and the phylogenetic tree was constructed using software MEGA4 (Kumar et al. 2001). The tree for all strains listed in table 1 is displayed in FIG. 1.

[0148]The 16S rDNA sequences of all strains listed in table 1 have 99% percent identity to t...

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Abstract

The present technology pertains to improved methods for the conversion of lignocellulosic biomass material to bio-fuels using novel isolated cellulolytic extreme thermophilic bacterial cells belonging to the genus Caldicellulosiruptor, and mutants thereof.

Description

FIELD OF THE DISCLOSURE[0001]The present disclosure pertains to improved methods for the conversion of lignocellulosic biomass material to biofuels using novel isolated cellulolytic extreme thermophilic bacterial cells belonging to the genus Caldicellulosiruptor, and mutants thereof.BACKGROUND[0002]Biofuel can be broadly defined as solid, liquid, or gas fuel derived from recently dead biological material. The derivation of biofuel from recently dead biological material distinguishes it from fossil fuels, which are derived from long dead biological material. Biofuel can be theoretically produced from any biological carbon source, but a common source of biofuel is photosynthetic plants. Many different plants and plant-derived materials may be used for biofuel manufacture. One strategy for producing biofuel involves growing crops high in either sugar (e.g., sugar cane, sugar beet, and sweet sorghum) or starch (e.g., corn / maize), and then using yeast fermentation to produce ethyl alcoho...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P7/10
CPCC12P7/10C12N1/20C12P3/00C12P7/065C12P7/54C12P7/56C12N1/205C12R2001/01Y02E50/10C12P7/40C12P7/62C12P2201/00
Inventor CURVERS, SIMONSVETLICHNYI, VITALY
Owner DIREVO INDAL BIOTECH