TABLE 2

Biochemical properties of recombinant native EcAAO and evolved AAO (FX7 mutant)

Biochemical propertiesEcAAOFX7 mutant
Molecular mass (Da)a
    SDS-PAGE65,000120,000
    MALDI-TOF-MS61,847c61,485
    Amino acid composition61,08860,934
Glycosylation degree (%)50
Thermal stability (T50 [°C])47.564.3
pI3.94.3
N-terminal end sequenceMADFDYVVGdADFDYVVVG
Sp act (U/mg)b7424
Secretion level (mg/liter)2
  • a Masses were estimated based on SDS-PAGE, MALDI-TOF-MS after deglycosylation with PNGase F, or the amino acid composition.

  • b That is, the specific activity for p-methoxybenzyl alcohol.

  • c This information was obtained from reference 17.

  • d Met1 (M) was added to the N-terminal end for cloning in E. coli.