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AssigneeUniversity of Virginia Patent Foundation
InventorsSimonne Guenette, Mark Kester, Keith Kozminski, Alec Brewer, Kobe Rogers, Annabel Wright, Aarati Pokharel
Estimating the term…
Provided are nucleic acids and vectors that collectively encode various gene products related to converting styrene to polyhydroxybutyrate (PHB). In some embodiments, the nucleic acids and vectors collectively encode a styrene monooxygenase polypeptide, a flavin reductase polypeptide, a styrene-oxide isomerase polypeptide, and a phenylacetaldehyde dehydrogenase polypeptide, an acetyl-CoA C-acetyltransferase polypeptide, a 3-ketoacyl-ACP reductase polypeptide, a class I poly(R)-hydroxyalkanoic acid synthase polypeptide, and optionally an influx porin polypeptide. Also provided are systems and methods for producing PHB from styrene, methods and systems for remediating polystyrene waste. In some embodiments, the systems are in vivo systems.
An in vivo system for converting styrene to polyhydroxybutyrate (PHB) in a bacterial culture, the system comprising a bacterium that comprises one or more plasmids, the one or more plasmids collectively encoding a styrene monooxygenase, a flavin reductase, a styrene-oxide isomerase, a phenylacetaldehyde dehydrogenase, an acetyl-CoA C-acetyltransferase, a 3-ketoacyl-ACP reductase, and a class I poly(R)-hydroxyalkanoic acid synthase, and optionally an influx porin.
The in vivo system of any of the preceding claims, wherein the first plasmid comprises a single terminator 3′ to the first, second, third, and fourth coding sequences, optionally wherein the single terminator comprises a nucleotide sequence that is selected from the group consisting of SEQ ID NOs:
The in vivo system of any of the preceding claims, wherein the second plasmid comprises a single terminator 3′ to the fifth, sixth, and seventh coding sequences, and a double terminator 3′ to the eighth coding sequence, if present, or both a single terminator 3′ to the fifth, sixth, and seventh coding sequences and a double terminator 3′ to the eighth coding sequence, if present.
The in vivo system of any of the preceding claims, further comprising a medium in which to culture the cell.
The in vivo system of any of the preceding claims, wherein the in vivo system further comprises a partitioning agent that enhances partitioning of styrene into medium in which the cell is growing.
The in vivo system of any of the preceding claims, wherein the cell is an Escherichia coli (E. coli ) bacterium, optionally an bacterium of the strain E. coli W or the strain E. coli TG1.
The in vivo system of any of the preceding claims, wherein one or more of the first-eighth coding sequences are modified to encode an epitope tag.
A method for remediating polystyrene (PS) waste, the method comprising:
An in vivo system for remediating polystyrene (PS) waste, the system comprising one or more bacteria comprising:
A plasmid comprising:
A plasmid comprising:
A nucleic acid comprising, consisting essentially of, or consisting of a nucleotide sequence of any one of SEQ ID NOs:
A vector encoding two or more of a styrene monooxygenase, a flavin reductase, a styrene-oxide isomerase, a phenylacetaldehyde dehydrogenase, an acetyl-CoA C-acetyltransferase, a 3-ketoacyl-ACP reductase, and a class I poly(R)-hydroxyalkanoic acid synthase, and optionally an influx porin.
A multi vector system, wherein:
Dependencies are inferred from the claim text.