Biochemical characterization of two haloalkane dehalogenases

DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea

Lauren Carlucci, Edward Zhou, Vladimir N. Malashkevich, Steven C. Almo, Emily C. Mundorff

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Two putative haloalkane dehalogenases (HLDs) of the HLD-I subfamily, DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea, have been identified based on sequence comparisons with functionally characterized HLD enzymes. The two genes were synthesized, functionally expressed in E. coli and shown to have activity toward a panel of haloalkane substrates. DsaA has a moderate activity level and a preference for long (greater than 3 carbons) brominated substrates, but little activity toward chlorinated alkanes. DccA shows high activity with both long brominated and chlorinated alkanes. The structure of DccA was determined by X-ray crystallography and was refined to 1.5 Å resolution. The enzyme has a large and open binding pocket with two well-defined access tunnels. A structural alignment of HLD-I subfamily members suggests a possible basis for substrate specificity is due to access tunnel size.

Original languageEnglish (US)
JournalProtein Science
DOIs
StateAccepted/In press - 2016

Fingerprint

haloalkane dehalogenase
Caulobacter crescentus
Alkanes
Tunnels
Substrates
X ray crystallography
X Ray Crystallography
Enzymes
Substrate Specificity
Escherichia coli
Carbon
Genes

Keywords

  • Caulobacter crescentus
  • DccA
  • Haloalkane dehalogenase
  • HLD-I subfamily
  • Saccharomonospora azurea
  • Substrate specificity

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Biochemical characterization of two haloalkane dehalogenases : DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea. / Carlucci, Lauren; Zhou, Edward; Malashkevich, Vladimir N.; Almo, Steven C.; Mundorff, Emily C.

In: Protein Science, 2016.

Research output: Contribution to journalArticle

@article{dda87f8582eb4b11ac5dfea659a24e6b,
title = "Biochemical characterization of two haloalkane dehalogenases: DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea",
abstract = "Two putative haloalkane dehalogenases (HLDs) of the HLD-I subfamily, DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea, have been identified based on sequence comparisons with functionally characterized HLD enzymes. The two genes were synthesized, functionally expressed in E. coli and shown to have activity toward a panel of haloalkane substrates. DsaA has a moderate activity level and a preference for long (greater than 3 carbons) brominated substrates, but little activity toward chlorinated alkanes. DccA shows high activity with both long brominated and chlorinated alkanes. The structure of DccA was determined by X-ray crystallography and was refined to 1.5 {\AA} resolution. The enzyme has a large and open binding pocket with two well-defined access tunnels. A structural alignment of HLD-I subfamily members suggests a possible basis for substrate specificity is due to access tunnel size.",
keywords = "Caulobacter crescentus, DccA, Haloalkane dehalogenase, HLD-I subfamily, Saccharomonospora azurea, Substrate specificity",
author = "Lauren Carlucci and Edward Zhou and Malashkevich, {Vladimir N.} and Almo, {Steven C.} and Mundorff, {Emily C.}",
year = "2016",
doi = "10.1002/pro.2895",
language = "English (US)",
journal = "Protein Science",
issn = "0961-8368",
publisher = "Cold Spring Harbor Laboratory Press",

}

TY - JOUR

T1 - Biochemical characterization of two haloalkane dehalogenases

T2 - DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea

AU - Carlucci, Lauren

AU - Zhou, Edward

AU - Malashkevich, Vladimir N.

AU - Almo, Steven C.

AU - Mundorff, Emily C.

PY - 2016

Y1 - 2016

N2 - Two putative haloalkane dehalogenases (HLDs) of the HLD-I subfamily, DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea, have been identified based on sequence comparisons with functionally characterized HLD enzymes. The two genes were synthesized, functionally expressed in E. coli and shown to have activity toward a panel of haloalkane substrates. DsaA has a moderate activity level and a preference for long (greater than 3 carbons) brominated substrates, but little activity toward chlorinated alkanes. DccA shows high activity with both long brominated and chlorinated alkanes. The structure of DccA was determined by X-ray crystallography and was refined to 1.5 Å resolution. The enzyme has a large and open binding pocket with two well-defined access tunnels. A structural alignment of HLD-I subfamily members suggests a possible basis for substrate specificity is due to access tunnel size.

AB - Two putative haloalkane dehalogenases (HLDs) of the HLD-I subfamily, DccA from Caulobacter crescentus and DsaA from Saccharomonospora azurea, have been identified based on sequence comparisons with functionally characterized HLD enzymes. The two genes were synthesized, functionally expressed in E. coli and shown to have activity toward a panel of haloalkane substrates. DsaA has a moderate activity level and a preference for long (greater than 3 carbons) brominated substrates, but little activity toward chlorinated alkanes. DccA shows high activity with both long brominated and chlorinated alkanes. The structure of DccA was determined by X-ray crystallography and was refined to 1.5 Å resolution. The enzyme has a large and open binding pocket with two well-defined access tunnels. A structural alignment of HLD-I subfamily members suggests a possible basis for substrate specificity is due to access tunnel size.

KW - Caulobacter crescentus

KW - DccA

KW - Haloalkane dehalogenase

KW - HLD-I subfamily

KW - Saccharomonospora azurea

KW - Substrate specificity

UR - http://www.scopus.com/inward/record.url?scp=84959158195&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84959158195&partnerID=8YFLogxK

U2 - 10.1002/pro.2895

DO - 10.1002/pro.2895

M3 - Article

JO - Protein Science

JF - Protein Science

SN - 0961-8368

ER -