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Applied and Environmental Microbiology, December 1998, p. 4809-4815, Vol. 64, No. 12
Nestlé Research Center,
Received 12 June 1998/Accepted 22 September 1998
The koji mold Aspergillus oryzae secretes a prolyl
dipeptidyl peptidase (DPPIV) when the fungus is cultivated in a medium containing wheat gluten as the sole nitrogen and carbon source (MMWG).
We cloned and sequenced the DPPIV gene from an A. oryzae library by using the A. fumigatus dppIV
gene as a probe. Reverse transcriptase PCR experiments showed that the
A. oryzae dppIV gene consists of two exons, the first
of which is only 6 bp long. The gene encodes an 87.2-kDa polypeptide
chain which is secreted into the medium as a 95-kDa glycoprotein.
Introduction of this gene into A. oryzae leads to
overexpression of prolyl dipeptidyl peptidase activity, while
disruption of the gene abolishes all prolyl dipeptidyl peptidase
activity in MMWG. The dppIV null mutants did not exhibit
any change in phenotype other than the absence of prolyl dipeptidyl
peptidase activity, suggesting that this activity is not essential.
This loss of activity diminished the number of dipeptides and increased
the number of larger peptides present in the MMWG culture broth. These
effects were reversed by the addition of purified, recombinant DPPIV
from the methylotrophic yeast expression vector Pichia
pastoris. Our results suggest that the DPPIV enzyme may be of
importance in industrial hydrolysis of what gluten-based
substrates, which are rich in Pro residues.
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Characterization of the Prolyl Dipeptidyl Peptidase Gene
(dppIV) from the Koji Mold Aspergillus
oryzae
*
Corresponding author. Mailing address: Service de
Dermatologie, Laboratoire de Mycologie, BT422, Centre Hospitalier
Universitaire Vaudois, 1011 Lausanne, Switzerland. Phone: 41 21 314 0376. Fax: 41 21 314 0378. E-mail:
Michel.Monod{at}chuv.hospvd.ch.
Applied and Environmental Microbiology, December 1998, p. 4809-4815, Vol. 64, No. 12
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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