Methods for biosynthesis of isobutene
AssigneeInvista North America S.A.R.L.
InventorsAlex Van Eck Conradie, Adriana Leonora Botes
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Abstract
The document provides methods for biosynthesizing isobutene using one or more isolated enzymes such as one or more of a hydratase such as an enzyme classified under EC 4.2.1.- and a decarboxylating thioesterase, or using recombinant host cells expressing one or more such enzymes.
Claims17
A method for synthesizing isobutene, said method comprising enzymatically converting 3-methyl-but-2-enoyl-CoA to form to 3-methyl-3-hydroxy-butanoyl-CoA using a R-specific enoyl-CoA hydratase with at least 90% sequence identity to SEQ ID NO:1, 2 or 3 and which is capable of converting 3-methyl-but-2-enoyl-CoA to 3-methyl-3-hydroxy-butanoyl-CoA, enzymatically converting 3-methyl-3-hydroxy-butanoyl-CoA to 3-methyl-3-sulphoryl-butanoyl-ACP using a CoA:ACP 3-hydroxyacyltransferase or phosphopantetheinyl transferase capable of converting 3-methyl-3-hydroxy-butanoyl-CoA to 3-methyl-3-hydroxy-butanoyl-ACP and a sulfotransferase capable of converting 3-methyl-3-hydroxy-butanoyl-ACP to 3-methyl-3-sulphoryl-butanoyl-ACP, and enzymatically converting 3-methyl-3-sulphoryl-butanoyl-ACP to isobutene using a decarboxylating thioesterase which has at least 90% sequence identity to Lyngbya majuscula chain A, thioesterase from the Curacin biosynthetic pathway comprising SEQ ID NO:4 and is capable of converting 3-methyl-3-sulphoryl-butanoyl-ACP to isobutene or a decarboxylating thioesterase capable of converting 3-methyl-3-sulphoryl-butanoyl-ACP to isobutene derived from Pseudomonas entomophila, Haliangium ochraceum, Synechococcus, or Cyanothece.
Dependencies are inferred from the claim text.