Enzymatic preparation of 5-hydroxy-L-proline, N-Cbz-5-hydroxy-L-proline, and N-Boc-5-hydroxy-L-proline from (α-N-protected)-L-ornithine using a transaminase or an amine oxidase

Enzyme Microb Technol. 2011 May 6;48(6-7):445-53. doi: 10.1016/j.enzmictec.2011.03.007. Epub 2011 Mar 26.

Abstract

N-Cbz-4,5-dehydro-L-prolineamide or N-Boc-4,5-dehydro-L-prolineamide are alternative key intermediates for the synthesis of saxagliptin, a dipeptidyl peptidase IV (DPP4) inhibitor recently approved for treatment of type 2 diabetes mellitus. An efficient biocatalytic method was developed for conversion of L-ornithine, N-α-benzyloxycarbonyl (Cbz)-L-ornthine, and N-α-tert-butoxycarbonyl (Boc)-L-ornithine to 5-hydroxy-L-proline, N-Cbz-5-hydroxy-L-proline, and N-Boc-5-hydroxy-L-proline, respectively. Rec. Escherichia coli expressing lysine-ε-aminotransferase and rec Pichia pastoris expressing L-ornithine oxidase were used for these conversions. N-Cbz-5-hydroxy-L-proline, and N-Boc-5-hydroxy-L-proline were chemically converted to key intermediates N-Cbz-4,5-dehydro-L-prolineamide and N-Boc-4,5-dehydro-L-prolineamide, respectively.

MeSH terms

  • Adamantane / analogs & derivatives*
  • Amino Acid Sequence
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Biocatalysis
  • Chromatography, High Pressure Liquid
  • Dipeptides / biosynthesis*
  • Escherichia coli / enzymology
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism*
  • Genes, Fungal
  • Hypoglycemic Agents
  • L-Amino Acid Oxidase / genetics
  • L-Amino Acid Oxidase / isolation & purification
  • L-Amino Acid Oxidase / metabolism*
  • L-Lysine 6-Transaminase / metabolism*
  • Molecular Sequence Data
  • Ornithine / analogs & derivatives
  • Ornithine / metabolism*
  • Pichia / enzymology
  • Pichia / isolation & purification
  • Proline / analogs & derivatives*
  • Proline / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Soil Microbiology
  • Sphingomonas / enzymology
  • Sphingomonas / genetics

Substances

  • Bacterial Proteins
  • Dipeptides
  • Fungal Proteins
  • Hypoglycemic Agents
  • Recombinant Fusion Proteins
  • Proline
  • saxagliptin
  • Ornithine
  • L-Amino Acid Oxidase
  • L-Lysine 6-Transaminase
  • Adamantane