In silico screening of 393 mutants facilitates enzyme engineering of amidase activity in CalB

Martin Robert Hediger; Luca De Vico; Julie Bille Rannes; Christian Jäckel; Werner Besenmatter; Allan Svendsen; Jan Halborg Jensen

doi:10.7717/peerj.145

In silico screening of 393 mutants facilitates enzyme engineering of amidase activity in CalB

Martin Robert Hediger, Luca De Vico, Julie Bille Rannes, Christian Jäckel, Werner Besenmatter, Allan Svendsen, Jan Halborg Jensen

Department of Chemistry

6 Citations (Scopus)

943 Downloads (Pure)

Abstract

Our previously presented method for high throughput computational screening of mutant activity (Hediger et al., 2012) is benchmarked against experimentally measured amidase activity for 22 mutants of Candida antarctica lipase B (CalB). Using an appropriate cutoff criterion for the computed barriers, the qualitative activity of 15 out of 22 mutants is correctly predicted. The method identifies four of the six most active mutants with ≥3-fold wild type activity and seven out of the eight least active mutants with ≤0.5-fold wild type activity. The method is further used to screen all sterically possible (386) double-, triple- and quadruple-mutants constructed from the most active single mutants. Based on the benchmark test at least 20 new promising mutants are identified.

Original language	English
Article number	e145
Journal	PeerJ
Volume	1
Number of pages	15
DOIs	https://doi.org/10.7717/peerj.145
Publication status	Published - 2013

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title = "In silico screening of 393 mutants facilitates enzyme engineering of amidase activity in CalB",

abstract = "Our previously presented method for high throughput computational screening of mutant activity (Hediger et al., 2012) is benchmarked against experimentally measured amidase activity for 22 mutants of Candida antarctica lipase B (CalB). Using an appropriate cutoff criterion for the computed barriers, the qualitative activity of 15 out of 22 mutants is correctly predicted. The method identifies four of the six most active mutants with ≥3-fold wild type activity and seven out of the eight least active mutants with ≤0.5-fold wild type activity. The method is further used to screen all sterically possible (386) double-, triple- and quadruple-mutants constructed from the most active single mutants. Based on the benchmark test at least 20 new promising mutants are identified.",

author = "Hediger, {Martin Robert} and {De Vico}, Luca and Rannes, {Julie Bille} and Christian J{\"a}ckel and Werner Besenmatter and Allan Svendsen and Jensen, {Jan Halborg}",

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T1 - In silico screening of 393 mutants facilitates enzyme engineering of amidase activity in CalB

AU - Hediger, Martin Robert

AU - De Vico, Luca

AU - Rannes, Julie Bille

AU - Jäckel, Christian

AU - Besenmatter, Werner

AU - Svendsen, Allan

AU - Jensen, Jan Halborg

PY - 2013

Y1 - 2013

N2 - Our previously presented method for high throughput computational screening of mutant activity (Hediger et al., 2012) is benchmarked against experimentally measured amidase activity for 22 mutants of Candida antarctica lipase B (CalB). Using an appropriate cutoff criterion for the computed barriers, the qualitative activity of 15 out of 22 mutants is correctly predicted. The method identifies four of the six most active mutants with ≥3-fold wild type activity and seven out of the eight least active mutants with ≤0.5-fold wild type activity. The method is further used to screen all sterically possible (386) double-, triple- and quadruple-mutants constructed from the most active single mutants. Based on the benchmark test at least 20 new promising mutants are identified.

AB - Our previously presented method for high throughput computational screening of mutant activity (Hediger et al., 2012) is benchmarked against experimentally measured amidase activity for 22 mutants of Candida antarctica lipase B (CalB). Using an appropriate cutoff criterion for the computed barriers, the qualitative activity of 15 out of 22 mutants is correctly predicted. The method identifies four of the six most active mutants with ≥3-fold wild type activity and seven out of the eight least active mutants with ≤0.5-fold wild type activity. The method is further used to screen all sterically possible (386) double-, triple- and quadruple-mutants constructed from the most active single mutants. Based on the benchmark test at least 20 new promising mutants are identified.

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DO - 10.7717/peerj.145

M3 - Journal article

C2 - 24010022

SN - 2167-8359

VL - 1

JO - PeerJ

JF - PeerJ

M1 - e145

ER -

In silico screening of 393 mutants facilitates enzyme engineering of amidase activity in CalB

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