Evolution of an organophosphate-degrading enzyme: A comparison of natural and directed evolution

H. Yang; P. D. Carr; S. Yu McLoughlin; J. W. Liu; I. Horne; X. Qiu; C. M.J. Jeffries; R. J. Russell; J. G. Oakeshott; D. L. Ollis

doi:10.1093/proeng/gzg013

Evolution of an organophosphate-degrading enzyme: A comparison of natural and directed evolution

H. Yang, P. D. Carr, S. Yu McLoughlin, J. W. Liu, I. Horne, X. Qiu, C. M.J. Jeffries, R. J. Russell, J. G. Oakeshott, D. L. Ollis^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

149 Citations (Scopus)

Abstract

Organophosphate-degrading enzyme from Agrobacterium radiobacter P230 (OPDA) is a recently discovered enzyme that degrades a broad range of organophosphates. It is very similar to OPH first isolated from Pseudomonas diminuta MG. Despite a high level of sequence identity, OPH and OPDA exhibit different substrate specificities. We report here the structure of OPDA and identify regions of the protein that are likely to give it a preference for substrates that have shorter alkyl substituents. Directed evolution was used to evolve a series of OPH mutants that had activities similar to those of OPDA. Mutants were selected for on the basis of their ability to degrade a number of substrates. The mutations tended to cluster in particular regions of the protein and in most cases, these regions were where OPH and OPDA had significant differences in their sequences.

Original language	English
Pages (from-to)	135-145
Number of pages	11
Journal	Protein Engineering
Volume	16
Issue number	2
DOIs	https://doi.org/10.1093/proeng/gzg013
Publication status	Published - 1 Feb 2003

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title = "Evolution of an organophosphate-degrading enzyme: A comparison of natural and directed evolution",

abstract = "Organophosphate-degrading enzyme from Agrobacterium radiobacter P230 (OPDA) is a recently discovered enzyme that degrades a broad range of organophosphates. It is very similar to OPH first isolated from Pseudomonas diminuta MG. Despite a high level of sequence identity, OPH and OPDA exhibit different substrate specificities. We report here the structure of OPDA and identify regions of the protein that are likely to give it a preference for substrates that have shorter alkyl substituents. Directed evolution was used to evolve a series of OPH mutants that had activities similar to those of OPDA. Mutants were selected for on the basis of their ability to degrade a number of substrates. The mutations tended to cluster in particular regions of the protein and in most cases, these regions were where OPH and OPDA had significant differences in their sequences.",

keywords = "Agrobacterium, Bioremediation, Carboxylated lysine, Directed evolution, Organophosphates, Phosphotriesterase",

author = "H. Yang and Carr, {P. D.} and McLoughlin, {S. Yu} and Liu, {J. W.} and I. Horne and X. Qiu and Jeffries, {C. M.J.} and Russell, {R. J.} and Oakeshott, {J. G.} and Ollis, {D. L.}",

year = "2003",

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doi = "10.1093/proeng/gzg013",

language = "English",

volume = "16",

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journal = "Protein Engineering",

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TY - JOUR

T1 - Evolution of an organophosphate-degrading enzyme

T2 - A comparison of natural and directed evolution

AU - Yang, H.

AU - Carr, P. D.

AU - McLoughlin, S. Yu

AU - Liu, J. W.

AU - Horne, I.

AU - Qiu, X.

AU - Jeffries, C. M.J.

AU - Russell, R. J.

AU - Oakeshott, J. G.

AU - Ollis, D. L.

PY - 2003/2/1

Y1 - 2003/2/1

N2 - Organophosphate-degrading enzyme from Agrobacterium radiobacter P230 (OPDA) is a recently discovered enzyme that degrades a broad range of organophosphates. It is very similar to OPH first isolated from Pseudomonas diminuta MG. Despite a high level of sequence identity, OPH and OPDA exhibit different substrate specificities. We report here the structure of OPDA and identify regions of the protein that are likely to give it a preference for substrates that have shorter alkyl substituents. Directed evolution was used to evolve a series of OPH mutants that had activities similar to those of OPDA. Mutants were selected for on the basis of their ability to degrade a number of substrates. The mutations tended to cluster in particular regions of the protein and in most cases, these regions were where OPH and OPDA had significant differences in their sequences.

AB - Organophosphate-degrading enzyme from Agrobacterium radiobacter P230 (OPDA) is a recently discovered enzyme that degrades a broad range of organophosphates. It is very similar to OPH first isolated from Pseudomonas diminuta MG. Despite a high level of sequence identity, OPH and OPDA exhibit different substrate specificities. We report here the structure of OPDA and identify regions of the protein that are likely to give it a preference for substrates that have shorter alkyl substituents. Directed evolution was used to evolve a series of OPH mutants that had activities similar to those of OPDA. Mutants were selected for on the basis of their ability to degrade a number of substrates. The mutations tended to cluster in particular regions of the protein and in most cases, these regions were where OPH and OPDA had significant differences in their sequences.

KW - Agrobacterium

KW - Bioremediation

KW - Carboxylated lysine

KW - Directed evolution

KW - Organophosphates

KW - Phosphotriesterase

UR - http://www.scopus.com/inward/record.url?scp=0037314857&partnerID=8YFLogxK

U2 - 10.1093/proeng/gzg013

DO - 10.1093/proeng/gzg013

M3 - Article

SN - 0269-2139

VL - 16

SP - 135

EP - 145

JO - Protein Engineering

JF - Protein Engineering

IS - 2

ER -

Evolution of an organophosphate-degrading enzyme: A comparison of natural and directed evolution

Abstract

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