Biological production of ß-lactones
AssigneeRegents of the University of Minnesota
InventorsSerina L. Robinson, James K. Christenson, Lawrence P. Wackett
Life of this patent
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Abstract
Methods of using β-lactone biosynthetic enzyme genes and host cells expressing one or more of those genes, e.g., heterologous expression, are provided.
Claims21
A method to prepare β-lactones in vitro, comprising:
- combining one or more acyl CoA substrates, one or more activated acyl substrates, one or more carboxylic acid substrates, or one or more fatty acid substrates with OleA or a homolog thereof, OleC or a homolog thereof, and OleD or a homolog thereof but not OleB, under conditions that yield one or more oxetan-2-ones.
A method for altering the ratio of trans lactones in a mixture of lactones, comprising:
- combining mixed diastereomers of an oxetan-2-one with OleA or a homolog thereof, OleD or a homolog thereof and OleC or a homolog thereof, so as to yield a mixture with an altered amount of trans-β-lactones.
A method to identify β-lactone synthetase activity, comprising:
- combining at room temperature and a pH of about 6 to about 8, a sample suspected of having β-lactone synthetase and a dialkene, a dialkyne or a compound with an alkene and alkene group, so as to yield a mixture;
- and
- detecting in the mixture a change in UV absorbance over time, wherein a change in absorbance is indicative of the presence or amount of a β-lactone synthetase.
A host cell comprising a genome augmented with a nucleic acid encoding OleA or a homolog thereof, a nucleic acid encoding OleC or a homolog thereof and a nucleic acid encoding OleD or a homolog, thereof, but which lacks OleB activity, wherein the host cell is heterologous to one or more of the OleA or homolog thereof, the OleC or homolog therof, the OleD or the homolog thereof or a host cell comprising a genome expressing a heterologous OleC.
Dependencies are inferred from the claim text.