Showing papers in "Acta Crystallographica in 2013"
Journal Article•
TL;DR: The two LinB variants presented here, LinB32 and LinB70, were constructed with the goal of studying the effect of mutations on enzyme functionality and suggest that the mutations, although deep inside the molecule, can still affect the protein crystallizability.
Abstract: Haloalkane dehalogenases are microbial enzymes that convert a
broad range of halogenated aliphatic compounds to their
corresponding alcohols by the hydrolytic mechanism. These
enzymes play an important role in the biodegradation of various
environmental pollutants. Haloalkane dehalogenase LinB isolated
from a soil bacterium Sphingobium japonicum UT26 has a
relatively broad substrate specificity and can be applied in
bioremediation and biosensing of environmental pollutants. The
LinB variants presented here, LinB32 and LinB70, were
constructed with the goal of studying the effect of mutations
on enzyme functionality. In the case of LinB32 (L117W), the
introduced mutation leads to blocking of the main tunnel
connecting the deeply buried active site with the surrounding
solvent. The other variant, LinB70 (L44I, H107Q), has the
second halide-binding site in a position analogous to that in
the related haloalkane dehalogenase DbeA from Bradyrhizobium
elkanii USDA94. Both LinB variants were successfully
crystallized and full data sets were collected for native
enzymes as well as their complexes with the substrates
1,2-dibromoethane (LinB32) and 1-bromobutane (LinB70) to
resolutions ranging from 1.6 to 2.8 A. The two mutants
crystallize differently from each other, which suggests that
the mutations, although deep inside the molecule, can still
affect the protein crystallizability.
6 citations
Journal Article•
TL;DR: In this paper, two newly isolated psychrophilic haloalkane dehalogenases, DpcA from Psychiatricrobacter cryohalolentis K5 and DmxA from Marinobacter sp.ELB17, were purified and used for crystallization experiments.
Abstract: Haloalkane dehalogenases are hydrolytic enzymes with a broad
range of potential practical applications such as
biodegradation, biosensing, biocatalysis and cellular imaging.
Two newly isolated psychrophilic haloalkane dehalogenases
exhibiting interesting catalytic properties, DpcA from
Psychrobacter cryohalolentis K5 and DmxA from Marinobacter sp.
ELB17, were purified and used for crystallization experiments.
After the optimization of crystallization conditions, crystals
of diffraction quality were obtained. Diffraction data sets
were collected for native enzymes and complexes with selected
ligands such as 1-bromohexane and 1,2-dichloroethane to
resolutions ranging from 1.05 to 2.49 A.
6 citations