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Apparatus for rapid biohydrogen phenotypic screening of microorganisms using a chemochromic sensor

a technology of biohydrogen phenotypic screening and chemochromic sensor, which is applied in the direction of biomass after-treatment, analysis using chemical indicators, instruments, etc., can solve the problems of limiting the commercial application of this process, and affecting the phenotypic screening rate of microorganisms

Inactive Publication Date: 2001-11-15
ALLIANCE FOR SUSTAINABLE ENERGY
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Problems solved by technology

While hydrogenase-catalyzed hydrogen photoproduction has potential as an efficient energy source, one of the major obstacles which currently limits any commercial application of this process is the deactivation of the hydrogenase enzyme in the presence of oxygen, produced in the water-splitting process of photosynthesis.
However, this assay for oxygen-tolerant hydrogen photo-production is very costly, time consuming, and has been the prime rate limiting factor in the identification and selection of more desirable mutant organisms.
However, these devices do not spatially resolve the point where the gas is produced in relation to the sample surface.
Therefore, these devices would not be useful to discriminate the specific location of a colony which produces hydrogen gas where the sample, to be screened, consists of many colonies of the organism.
While this patent provides an optical means for detecting hydrogen which could provide spatial resolution it does not provide the sensitivity (0.02% hydrogen concentration), rapid response rate (a few seconds), and economy in manufacture, which are desirable for rapidly screening colonies of hydrogen-producing organisms on a substrate.

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  • Apparatus for rapid biohydrogen phenotypic screening of microorganisms using a chemochromic sensor
  • Apparatus for rapid biohydrogen phenotypic screening of microorganisms using a chemochromic sensor
  • Apparatus for rapid biohydrogen phenotypic screening of microorganisms using a chemochromic sensor

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Embodiment Construction

[0034] Wild type C. reinhardtii (WT) was obtained from the University of Colorado at Boulder and a cell-wall-less strain of C. reinhardtii (cw15) was obtained from the Chlamydomonas Genetics Center, Duke University. Algal cell suspensions were grown photoautotrophically either in Sager's minimal medium (WT), or in a modified Sueoka'a high salt medium, as described in Ghirardi et. al., Development of an Efficient Algal H.sub.2-Producing System, Proceedings of the 1996 U.S. DOE Hydrogen Program Review, Vol. 1, 285-302, (1997). Suspension cultures were grown under illumination at 25.degree. C., using cool white fluorescent lights (8W / m.sup.2), and agitated with a bubble mixture of 1.7% CO.sub.2 in air. Plated colonies were prepared by centrifugation harvesting of cell suspensions at 2000 g, for 10 minutes, and inoculated on either 1.5% (WT), or 0.8% (cw15) agar gel in sterile plastic Petri dishes.

Mutageneses

[0035] In order to generate C. reinhardtii mutants to be used in the two select...

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Abstract

Provided is a system for identifying a hydrogen gas producing organism. The system includes a sensor film having a first layer comprising a transition metal oxide or oxysalt and a second layer comprising a hydrogen-dissociative catalyst metal, the first and second layers having an inner and an outer surface wherein the inner surface of the second layer is deposited on the outer surface of the first layer, and a substrate adjacent to the outer surface of the second layer, the organism isolated on the substrate.

Description

[0001] This is a 35 U.S.C. 121 divisional application of the co-pending 35 U.S.C. 111(a) application, Ser. No. 09 / 316,172, filed May 21, 1999, which claims the benefit, pursuant to 35 U.S.C. 119(e)(1), of an invention disclosed in the 35 U.S.C. 111(b) provisional application, serial number 60 / 086,313, filed May 21, 1998.REFERENCE TO A MICROFICHE APPENDIX.[0003] Not Applicable.[0004] 1. Field of the Invention[0005] This invention relates to chemochromic sensor films for use in rapidly screening isolate organisms capable of producing hydrogen.[0006] 2. Description of the Prior Art[0007] Light induced biological hydrogen production represents a potentially cost effective system for the production of renewable non-polluting energy. Photobiological hydrogen production is catalyzed by the nitrogenase and hydrogenase enzyme systems that are present in photosynthetic bacteria, cyanobacteria, and green algae. Green algae, such as Chlamydomonas reinhardtii, can photoevolve hydrogen only throu...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/04G01N31/22G01N33/00G01N33/497
CPCC12Q1/04C12Q2304/40G01N31/223G01N33/005G01N2033/4977
Inventor SEIBERT, MICHAELBENSON, DAVID K.FLYNN, TIMOTHY MICHAEL
Owner ALLIANCE FOR SUSTAINABLE ENERGY
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