Transaminase biocatalysts

  • US 9,133,445 B2
  • Filed: 10/20/2014
  • Issued: 09/15/2015
  • Est. Priority Date: 02/26/2009
  • Status: Active Grant
First Claim
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1. A recombinant polynucleotide sequence encoding a transaminase polypeptide comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:

  • 2, wherein the amino acid at position 223 of said amino acid sequence is proline, and the amino acid at position 122 of said amino acid sequence is isoleucine, methionine, valine, or histidine, and further wherein said polypeptide is capable of converting 4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-one (“

    ketoamide substrate”

    ) to (2R)-4-oxo-4-[3-(trifluoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]-1-(2,4,5-trifluorophenyl)butan-2-amine (“


    ) under a defined reaction condition in the presence of an amino group donor isopropylamine to levels of the product detectable by HPLC-UV at 210 nm, where the reaction condition comprises about 2 g/L ketoamide substrate, about 0.5 M isopropylamine, about 22°

    C., about pH 7.5, about 5% DMSO, about 100 μ

    M pyridoxal phosphate, and about 20 mg/mL of transaminase polypeptide.

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