Newly discovered bacterium in the family acetobacteraceae
Inventors
Holland, Steven M. • Greenberg, David E. • Zelazny, Adrian • Murray, Patrick
Assignees
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Publication Number
US-9051573-B2
Publication Date
2015-06-09
Expiration Date
2027-03-28
Abstract
Provided is an isolated novel Gram-negative bacterium, wherein the bacterium is an aerobic, facultative methylotroph that produces colonies that are yellow pigmented, wherein the bacterium can use methanol as a sole carbon source and can oxidize glucose and ethanol into acid. Also provided are novel purified polypeptides and isolated nucleic acids from the bacterium. Further provided are methods of using the bacterium and the purified polypeptides to degrade organic material and for use in biofuel cells.
Core Innovation
This invention relates to an isolated novel Gram-negative bacterium that is an aerobic, facultative methylotroph producing yellow pigmented colonies. The bacterium can utilize methanol as its sole carbon source and has the ability to oxidize glucose and ethanol into acid. The organism, designated Granulibacter bethesdensis, was isolated from lymph nodes of a subject diagnosed with chronic granulomatous disease. The invention extends to purified polypeptides and isolated nucleic acids derived from the bacterium, and methods of using the bacterium and these polypeptides to degrade organic materials and for applications in biofuel cells.
The background identifies the problem that industrial activities generate organic wastes such as methanol, formaldehyde, and ethanol, the treatment of which is costly and hinders market expansion. There is a need for a biomass degrading and treatment system capable of converting these organic wastes into non-toxic end-products, thereby reducing treatment costs, preventing environmental pollution, and improving soil quality. Furthermore, producing and purifying the enzymes responsible for degradation would allow treatment in situations where the presence of live bacteria is unnecessary, and these enzymes could be applied in biofuel cells to generate electrical energy from organic materials.
The summary of the invention details that the isolated bacterium is capable of degrading organic materials including methanol, formaldehyde, and ethanol, into non-toxic products such as carbon dioxide, hydrogen ions, and acetic acid. The invention encompasses methods of degrading organic materials using the bacterium, culturing the bacterium under conditions effective to promote growth, and integrating the bacterium or its enzyme components in biofuel cells to convert organic substrates into electrical energy. The bacterium and its components provide a new genus and species within the family Acetobacteraceae with biochemical pathways for efficient degradation and oxidation reactions valuable for environmental and energy applications.
Claims Coverage
The patent includes a single independent claim defining a biofuel cell incorporating the novel bacterium with specific functional and genetic features.
Biofuel cell comprising a novel Gram-negative methylotroph bacterium
A biofuel cell comprising a cathode, an anode, a conductive medium, and an ion exchange membrane, wherein a bacterial catalyst surrounds the anode. The bacterial catalyst includes an isolated Gram-negative bacterium that is aerobic, facultative methylotroph producing yellow pigmented colonies, can use methanol as sole carbon source, oxidize glucose and ethanol into acid, and comprises a 16S rRNA nucleic acid identified as SEQ ID NO:2.
Identity and deposition of the bacterium
The bacterium is designated Granulibacter bethesdensis and is deposited under ATCC Accession No. BAA-1260.
Capability to degrade methanol and ethanol
The bacterium can degrade methanol into carbon dioxide and hydrogen ions and degrade ethanol into acetic acid.
Comprehensive genome and polypeptide profile
The bacterium includes a genome identified as SEQ ID NO:1 (GenBank CP000394) and produces methanol dehydrogenase, capable of oxidizing methanol into formaldehyde.
Methanol dehydrogenase subunit compositions
Methanol dehydrogenase comprises a first subunit peptide that can be one of four peptides with amino acid sequences identified as SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6 and a second subunit peptide with amino acid sequence SEQ ID NO:7, encoded by respective nucleic acids.
Production of a cascade of enzymes degrading methanol to carbon dioxide
The bacterium produces a series of enzymes including formaldehyde-activating enzyme (SEQ ID NO:13), methylenetetrahydromethanopterin dehydrogenase (SEQ ID NO:15), N5N10-methenyltetrahydromethanopterin cyclohydrolase (SEQ ID NO:17), formylmethanofuran-tetrahydromethanopterin formyltransferase (SEQ ID NO:19), tungsten-containing formylmethanofuran dehydrogenase subunits (SEQ ID NO:21, 22, 23), methylenetetrahydrofolate dehydrogenase (SEQ ID NO:27), methylenetetrahydrofolate cyclohydrolase (SEQ ID NO:29), formate-tetrahydrofolate ligase (SEQ ID NO:31), formate dehydrogenase subunits (SEQ ID NO:33 to 37), aldehyde dehydrogenase subunits (SEQ ID NO:43 to 45), and alcohol dehydrogenase (SEQ ID NO:48), all involved in organic material degradation and biofuel generation.
The claims broadly cover a biofuel cell that incorporates the inventive bacterium Granulibacter bethesdensis with specified genetic and enzymatic features enabling the degradation of methanol, ethanol, and related substrates into acids and further into electrical energy, highlighting the bacterium's unique enzymatic composition and deposited genetic material.
Stated Advantages
Reduces waste treatment costs by providing an efficient method to degrade organic waste like methanol, formaldehyde, and ethanol.
Prevents pollution of the environment through biodegradation of organic pollutants into non-toxic end products.
Improves soil quality and farmland by converting organic materials into benign products.
Enables mass production of purified bacterium-derived enzymes for organic material degradation in environments where live bacteria are not required.
Allows use of purified enzymes or whole bacteria in biofuel cells to convert organic waste materials into electrical energy.
Documented Applications
Treatment and biodegradation of industrial organic wastes such as methanol, formaldehyde, and ethanol to produce non-toxic end-products.
Use in biofuel cells where the bacterium or its enzymes convert organic matter into electrical energy.
Production and purification of bacterium-derived polypeptide enzymes to degrade organic materials in spill cleanup or environmental detoxification scenarios where the bacterium presence is not needed.
Culturing the bacterium under specified conditions for growth and mass production.
Use of immunological methods for detecting the bacterium or its components in biological samples, for example lymph node cultures of patients.
Power generation in miniature biofuel cells applicable to devices such as cell phones, laptops, remote sensors, and in-body medical implants.
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