Enzymatic preparation of 2-hydroxy-2-methylcarboxylic acid comprises producing 3-hydroxycarboxylic acid in aqueous reaction solution, incubating the obtained solution and converting to 2-hydroxy-2-methylcarboxylic acid

摘要

Enzymatic preparation of 2-hydroxy-2-methylcarboxylic acid from 3-hydroxycarboxylic acid, comprises producing 3-hydroxycarboxylic acid in an aqueous reaction solution containing 3-hydroxycarboxylic acid-CoA-mutase-activity exhibiting unit, which possesses both 3-hydroxycarbonyl CoA-ester producing- and 3-hydroxycarbonyl-CoA-ester isomerizing activity; incubating the reaction solution; and subsequently converting to 2-hydroxy-2-methylcarboxylic acid as an acid or in form of its salts. Independent claims are included for: (1) preparation of 2-3C-unsaturated isoalkenic acid using 2-hydroxy-2-methylcarboxylic acid dehydrate; (2) preparation of methyl acrylic acid using 2-hydroxyisobutyric acid dehydrate; (3) a microorganism strain HCM-10-DSM 18028; (4) a nucleic acid molecule coding enzyme with an activity of cobalamin-dependent mutase such as: nucleic acid molecule of protein code comprises fully defined 548 amino acid sequence (SEQ ID NO: 2) and/or fully defined 123 amino acid sequence (SEQ ID NO: 4), as given in the specification; a nucleic acid molecule comprising 1644 bp nucleotide sequence (SEQ ID NO: 1) and/or 369 bp nucleotide sequence (SEQ ID NO: 3), as given in the specification; nucleic acid molecule with the above mentioned amino acid sequence or nucleotide sequence that are hybridized; and a nucleic acid molecule, whose nucleotide sequence based on the degeneration of the genetic code of the sequence of the mentioned nucleotide sequence or the hybridized nucleotide- or amino acid -sequence; (5) a protein with the activity of a cobalamin-dependent mutase codes the mentioned nucleic acid molecule; and (6) protein as heterodimer enzyme comprising amino acids sequence of (SEQ ID NO: 2) and (SEQ ID No: 4), which are at least 60% homologous to the protein.