Hydrogen production from microbial strains

Inactive Publication Date: 2012-08-30
UNIV OF WASHINGTON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0012]An analysis of strains of diverse species of photosynthetic bacteria using the present invention can reveal new hydrogen-producing enzymes, highlight the diversity of strategies that can be used by bacteria to regulate hydrogen production, and uncover new genes for enabling hydrogen-producing enzymes to function efficiently in whole cells.

Problems solved by technology

The current challenge, however, is to develop technologies for hydrogen production from these resources that are clean, efficient, and cost effective.
This difficult reaction requires large amounts of ATP and reductant and, therefore, does not represent an efficient method of hydrogen production in terms of commercial utility (Simpson & Burris, “A Nitrogen Pressure of 50 Atmospheres Does Not Prevent Evolution of Hydrogen by Nitrogenase,”Science.

Method used

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  • Hydrogen production from microbial strains
  • Hydrogen production from microbial strains
  • Hydrogen production from microbial strains

Examples

Experimental program
Comparison scheme
Effect test

example 1

Bacterial Strains and Growth Conditions

[0032]The bacterial strains and plasmids used are listed in Table 1.

TABLE 1Strains, Plasmids and PrimersStrain, plasmid,Reference, origin,or primerGenotype or phenotypeaor descriptionR. palustris strainsCGA009Wild type strain; spontaneous CmR derivative of CGA0011CGA570Spontaneous mutant able to grow with cyclohexanecarboxylateCGA009, thisin the presence of ammonium sulfate (NifA S213P)studyCGA571Spontaneous mutant able to grow with cyclohexanecarboxylateCGA009, thisin the presence of ammonium sulfate (NifA Q209P)studyCGA572Spontaneous mutant able to grow with cyclohexanecarboxylateCGA009, thisin the presence of ammonium sulfate (NifA, L212R)studyCGA574Spontaneous mutant able to grow with cyclohexanecarboxylateCGA009, thisin the presence of ammonium sulfate (NifA M202K)studyCGA757ΔnifA, 1332 bp deletedCGA009, thisstudyCGA581nifA* (Q209P)CGA757, thisstudyCGA584nifA* (M202K)CGA757, thisstudyE. coli strainsDH5αF−λ− recA1Δ(lacZYA-argF)U169hsdR17thi...

example 2

Strain Constructions

[0034]DNA fragments (˜3 kb) spanning nifA plus flanking regions from CGA571 and CGA574 were generated by PCR. The amplification products contained engineered XbaI cloning sites at both ends. These products were digested with XbaI and cloned into XbaI-digested pUC19 to generate pUC19-nifA *571, and pUC19-nifA*574. The Xba-I fragments were then cloned into pJQ200KS to generate pJQ200KS-nifA *571, and pJQ200KS-nifA*574. These constructs were mobilized from E. coli S17-1 into R. palustris CGA757 (ΔnifA, this strain cannot grow under nitrogen-fixing conditions) by conjugation. Colonies that contained plasmids that had undergone a single recombination to become inserted into the chromosome were identified by growth on PM plus Gm. These colonies were spread onto nitrogen-fixing agar medium supplemented with 10% sucrose and incubated anaerobically in order to select for strains that had undergone a double recombination to lose the sacB-containing vector. Gene replacement...

example 3

Description of the R. palustris Genechip

[0035]R. palustris custom designed GeneChip was manufactured by Affymetrix (Santa Clara, Calif.) with the following specifications: 99.8% of R. palustris predicted open reading frames genes were represented by unique probe sets (16 probe pairs and their corresponding mismatch for each). In addition, the GeneChip contains information for all intergenic regions larger than 150 bp.

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Abstract

The present invention is directed to a method of screening microbe strains capable of generating hydrogen. This method involves inoculating one or more microbes in a sample containing cell culture medium to form an inoculated culture medium. The inoculated culture medium is then incubated under hydrogen producing conditions. Once incubating causes the inoculated culture medium to produce hydrogen, microbes in the culture medium are identified as candidate microbe strains capable of generating hydrogen. Methods of producing hydrogen using one or more of the microbial strains identified as well as the hydrogen producing strains themselves are also disclosed.

Description

[0001]This application claims the benefit of U.S. Provisional Patent Application Ser. No. 60 / 868,233 filed Dec. 1, 2006, which is hereby incorporated by reference in its entirety.[0002]The subject matter of this application was made, at least in part, with funding received from the U.S. Department of Energy under grant DE-FG02-05ER64063 and the U.S. Department of Defense under grant W911NF-05-1-0176. The U.S. Government may have certain rights.FIELD OF THE INVENTION[0003]The present invention generally relates to the production of hydrogen from microbial strains.BACKGROUND OF THE INVENTION[0004]Hydrogen biofuel has the potential to solve a variety of challenges related to the global need for a clean and sustainable form of energy. Hydrogen can be produced from a variety of domestic resources including: fossil fuels such as natural gas and coal; renewable resources such as solar, wind, and biomass; or nuclear energy. The current challenge, however, is to develop technologies for hydr...

Claims

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Application Information

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IPC IPC(8): C12P3/00
CPCC12P3/00
InventorHARWOOD, CAROLINE S.REY, FEDERICO E.
OwnerUNIV OF WASHINGTON