Non-enzymatic and enzymatic hydrolysis of alkyl halides: A haloalkane dehalogenation enzyme evolved to stabilize the gas-phase transition state of an S(N)2 displacement reaction

Felice C Lightstone, Y. J. Zheng, A. H. Maulitz, T. C. Bruice

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Abstract

The semiempirical PM3 method, calibrated against ab initio HF/6-31+G(d) theory, has been used to elucidate the reaction of 1,2-dichloroethane (DCE) with the carboxylate of Asp-124 at the active site of haloalkane dehalogenase of Xanthobacter autothropicus. Asp-124 and 13 other amino acid side chains that make up the active site cavity (Glu-56, Trp-125, Phe-128, Phe-172, Trp- 175, Leu-179, Val219, Phe-222, Pro-223, Val-226, Leu-262, Leu-263, and His- 289) were included in the calculations. The three most significant observations of the present study are that: (i) the DCE substrate and Asp- 124 carboxylate, in the reactive ES complex, are present as an ion-molecule complex with a structure similar to that seen in the gas-phase reaction of AcO- with DCE; (ii) the structures of the transition states in the gas- phase and enzymatic reaction are much the same where the structure formed at the active site is somewhat exploded; and (iii) the enthalpies in going from ground states to transition states in the enzymatic and gas-phase reactions differ by only a couple kcal/mol. The dehalogenase derives its catalytic power from: (i) bringing the electrophile and nucleophile together in a low- dielectric environment in an orientation that allows the reaction to occur without much structural reorganization; (ii) desolvation; and (iii) stabilizing the leaving chloride anion by Trp-125 and Trp-175 through hydrogen bonding.

Original languageEnglish (US)
Pages (from-to)8417-8420
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number16
DOIs
StatePublished - 1997

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Phase Transition
Ethylene Dichlorides
Catalytic Domain
haloalkane dehalogenase
Hydrolysis
Gases
Xanthobacter
Enzymes
Hydrogen Bonding
Anions
Chlorides
Ions
Amino Acids

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

@article{377d78febb4b4bed948c22d2806f5fd3,
title = "Non-enzymatic and enzymatic hydrolysis of alkyl halides: A haloalkane dehalogenation enzyme evolved to stabilize the gas-phase transition state of an S(N)2 displacement reaction",
abstract = "The semiempirical PM3 method, calibrated against ab initio HF/6-31+G(d) theory, has been used to elucidate the reaction of 1,2-dichloroethane (DCE) with the carboxylate of Asp-124 at the active site of haloalkane dehalogenase of Xanthobacter autothropicus. Asp-124 and 13 other amino acid side chains that make up the active site cavity (Glu-56, Trp-125, Phe-128, Phe-172, Trp- 175, Leu-179, Val219, Phe-222, Pro-223, Val-226, Leu-262, Leu-263, and His- 289) were included in the calculations. The three most significant observations of the present study are that: (i) the DCE substrate and Asp- 124 carboxylate, in the reactive ES complex, are present as an ion-molecule complex with a structure similar to that seen in the gas-phase reaction of AcO- with DCE; (ii) the structures of the transition states in the gas- phase and enzymatic reaction are much the same where the structure formed at the active site is somewhat exploded; and (iii) the enthalpies in going from ground states to transition states in the enzymatic and gas-phase reactions differ by only a couple kcal/mol. The dehalogenase derives its catalytic power from: (i) bringing the electrophile and nucleophile together in a low- dielectric environment in an orientation that allows the reaction to occur without much structural reorganization; (ii) desolvation; and (iii) stabilizing the leaving chloride anion by Trp-125 and Trp-175 through hydrogen bonding.",
author = "Lightstone, {Felice C} and Zheng, {Y. J.} and Maulitz, {A. H.} and Bruice, {T. C.}",
year = "1997",
doi = "10.1073/pnas.94.16.8417",
language = "English (US)",
volume = "94",
pages = "8417--8420",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
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}

TY - JOUR

T1 - Non-enzymatic and enzymatic hydrolysis of alkyl halides

T2 - A haloalkane dehalogenation enzyme evolved to stabilize the gas-phase transition state of an S(N)2 displacement reaction

AU - Lightstone, Felice C

AU - Zheng, Y. J.

AU - Maulitz, A. H.

AU - Bruice, T. C.

PY - 1997

Y1 - 1997

N2 - The semiempirical PM3 method, calibrated against ab initio HF/6-31+G(d) theory, has been used to elucidate the reaction of 1,2-dichloroethane (DCE) with the carboxylate of Asp-124 at the active site of haloalkane dehalogenase of Xanthobacter autothropicus. Asp-124 and 13 other amino acid side chains that make up the active site cavity (Glu-56, Trp-125, Phe-128, Phe-172, Trp- 175, Leu-179, Val219, Phe-222, Pro-223, Val-226, Leu-262, Leu-263, and His- 289) were included in the calculations. The three most significant observations of the present study are that: (i) the DCE substrate and Asp- 124 carboxylate, in the reactive ES complex, are present as an ion-molecule complex with a structure similar to that seen in the gas-phase reaction of AcO- with DCE; (ii) the structures of the transition states in the gas- phase and enzymatic reaction are much the same where the structure formed at the active site is somewhat exploded; and (iii) the enthalpies in going from ground states to transition states in the enzymatic and gas-phase reactions differ by only a couple kcal/mol. The dehalogenase derives its catalytic power from: (i) bringing the electrophile and nucleophile together in a low- dielectric environment in an orientation that allows the reaction to occur without much structural reorganization; (ii) desolvation; and (iii) stabilizing the leaving chloride anion by Trp-125 and Trp-175 through hydrogen bonding.

AB - The semiempirical PM3 method, calibrated against ab initio HF/6-31+G(d) theory, has been used to elucidate the reaction of 1,2-dichloroethane (DCE) with the carboxylate of Asp-124 at the active site of haloalkane dehalogenase of Xanthobacter autothropicus. Asp-124 and 13 other amino acid side chains that make up the active site cavity (Glu-56, Trp-125, Phe-128, Phe-172, Trp- 175, Leu-179, Val219, Phe-222, Pro-223, Val-226, Leu-262, Leu-263, and His- 289) were included in the calculations. The three most significant observations of the present study are that: (i) the DCE substrate and Asp- 124 carboxylate, in the reactive ES complex, are present as an ion-molecule complex with a structure similar to that seen in the gas-phase reaction of AcO- with DCE; (ii) the structures of the transition states in the gas- phase and enzymatic reaction are much the same where the structure formed at the active site is somewhat exploded; and (iii) the enthalpies in going from ground states to transition states in the enzymatic and gas-phase reactions differ by only a couple kcal/mol. The dehalogenase derives its catalytic power from: (i) bringing the electrophile and nucleophile together in a low- dielectric environment in an orientation that allows the reaction to occur without much structural reorganization; (ii) desolvation; and (iii) stabilizing the leaving chloride anion by Trp-125 and Trp-175 through hydrogen bonding.

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U2 - 10.1073/pnas.94.16.8417

DO - 10.1073/pnas.94.16.8417

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JF - Proceedings of the National Academy of Sciences of the United States of America

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