Abstract
Discovery of new catalysts for demanding aqueous reactions is challenging. In here, we describe methodology for selection of catalytic phages by taking advantage of localized assembly of the product of the catalytic reaction that is screened for. A phage display library covering 109 unique dodecapeptide sequences is incubated with non-assembling precursors. Phages which are able to catalyze formation of the self-assembling reaction product (via amide condensation) acquire an aggregate of reaction product, enabling separation by centrifugation. The thus selected phages can be ampli-fied by infection of E.coli. These phages are shown to catalyze amide condensation and hydrolysis. Kinetic analysis shows a minor role for substrate binding. The approach enables discovery and mass-production of biocatalytic phages.
| Language | English |
|---|---|
| Pages | 15893–15896 |
| Number of pages | 4 |
| Journal | Journal of the American Chemical Society |
| Volume | 136 |
| Issue number | 45 |
| Early online date | 24 Oct 2014 |
| DOIs | |
| Publication status | Published - 2014 |
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Keywords
- catalysis
- biocatalysis
- esterase
- amidase
- phage display
- self-assembly
Cite this
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Discovery of catalytic phages by biocatalytic self-assembly. / Maeda, Yoshiaki; Javid, Nadeem; Duncan, Krystyna; Birchall, Louise; Gibson, Kirsty F; Cannon, Daniel; Kanetsuki, Yuka; Knapp, Charles W; Tuttle, Tell; Ulijn, Rein V.; Matsui, Hiroshi.
In: Journal of the American Chemical Society, Vol. 136, No. 45, 2014, p. 15893–15896.Research output: Contribution to journal › Article
TY - JOUR
T1 - Discovery of catalytic phages by biocatalytic self-assembly
AU - Maeda, Yoshiaki
AU - Javid, Nadeem
AU - Duncan, Krystyna
AU - Birchall, Louise
AU - Gibson, Kirsty F
AU - Cannon, Daniel
AU - Kanetsuki, Yuka
AU - Knapp, Charles W
AU - Tuttle, Tell
AU - Ulijn, Rein V.
AU - Matsui, Hiroshi
PY - 2014
Y1 - 2014
N2 - Discovery of new catalysts for demanding aqueous reactions is challenging. In here, we describe methodology for selection of catalytic phages by taking advantage of localized assembly of the product of the catalytic reaction that is screened for. A phage display library covering 109 unique dodecapeptide sequences is incubated with non-assembling precursors. Phages which are able to catalyze formation of the self-assembling reaction product (via amide condensation) acquire an aggregate of reaction product, enabling separation by centrifugation. The thus selected phages can be ampli-fied by infection of E.coli. These phages are shown to catalyze amide condensation and hydrolysis. Kinetic analysis shows a minor role for substrate binding. The approach enables discovery and mass-production of biocatalytic phages.
AB - Discovery of new catalysts for demanding aqueous reactions is challenging. In here, we describe methodology for selection of catalytic phages by taking advantage of localized assembly of the product of the catalytic reaction that is screened for. A phage display library covering 109 unique dodecapeptide sequences is incubated with non-assembling precursors. Phages which are able to catalyze formation of the self-assembling reaction product (via amide condensation) acquire an aggregate of reaction product, enabling separation by centrifugation. The thus selected phages can be ampli-fied by infection of E.coli. These phages are shown to catalyze amide condensation and hydrolysis. Kinetic analysis shows a minor role for substrate binding. The approach enables discovery and mass-production of biocatalytic phages.
KW - catalysis
KW - biocatalysis
KW - esterase
KW - amidase
KW - phage display
KW - self-assembly
U2 - 10.1021/ja509393p
DO - 10.1021/ja509393p
M3 - Article
VL - 136
SP - 15893
EP - 15896
JO - Journal of the American Chemical Society
T2 - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 45
ER -