Development of whole-cell biocatalyst for hydrolyzing high hydrophobic
pyrethroid pesticide by functional display of Aminopeptidase on the
surface of Saccharomyces cerevisiae
Abstract
In this study, we developed two new whole-cell biocatalysts by
immobilizing aminopeptidase (Aps) on the surface of yeast cells, using
N-terminal fusion and C-terminal fusion through lectin mediated display
system. After the two strains were cultured in the medium with galactose
as inducer for 48 hours, the activity of expressing Aps was at a high
level of 0.25 U/OD600/mL and 0.12 U/OD600/mL. The correct location of
Aps was confirmed by immunofluorescence analysis and flow cytometry.
Afterwards two whole cell catalysts could be reused with high stability
as it retained more than 70% of initial activity after ten repeated
batch reactions. Using β-cypermethrin (β-CP) as a substrate, the
effectiveness of two new whole-cell catalysts in the treatment of highly
hydrophobic organic pollutants was evaluated. The results showed that
when the concentration of β-CP was 200 mg·L -1, the
hydrolysis rates of the two whole cell catalysts were 33.16 μmol·L
-1·day -1 and 28.99 μmol·L
-1·day -1, and has the ability to
degrade a variety of pyrethroid pesticides. The β-CP residue in lettuce
and cherry tomatoes could be removed more than 70% under the conditions
of the Aga2N-Aps whole cell catalyst preparation dilution of 100 times.
This is the first report on the development of surface display Aps
biocatalyst, which can be used as an effective and renewable alternative
for the treatment of highly hydrophobic organic pollutants.