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Novosphingobium barchaimii sp. nov., isolated from hexachlorocyclohexane-contaminated soil.
TLDR
The results obtained from DNA-DNA hybridization and biochemical and physiological tests clearly distinguished the isolate from its closest phylogenetic neighbours, indicating that strain LL02(T) represents a novel species of the genus Novosphingobium. Expand
Exacerbation of substrate toxicity by IPTG in Escherichia coli BL21(DE3) carrying a synthetic metabolic pathway
TLDR
It is shown that induction with lactose, the natural inducer of Plac, dramatically lightens the burden without reducing the efficiency of the synthetic TCP degradation pathway, suggesting that lactose may be a better inducer than IPTG for the expression of heterologous pathways in E. coli BL21(DE3). Expand
Degradation of β-Hexachlorocyclohexane by Haloalkane Dehalogenase LinB from Sphingomonas paucimobilis UT26
TLDR
It is demonstrated that the haloalkane dehalogenase LinB, reported earlier to mediate the second step in the degradation of γ-HCH in Sphingomonas paucimobilis UT26, metabolizes β-H CH to produce 2-3,4,5,6-pentachlorocyclohexanol. Expand
Strategies for Stabilization of Enzymes in Organic Solvents
One of the major barriers to the use of enzymes in industrial biotechnology is their insufficient stability under processing conditions. The use of organic solvent systems instead of aqueous mediaExpand
Engineering enzyme stability and resistance to an organic cosolvent by modification of residues in the access tunnel.
TLDR
Protein crystallography and molecular dynamics revealed that the tunnel residue packing is a key determinant of protein stability and the active-site accessibility for co-solvent molecules. Expand
Haloalkane dehalogenases: Biotechnological applications
TLDR
This review discusses the application of HLDs in the context of the biochemical properties of individual enzymes and proposes that strategies based on protein engineering and isolation of novel H LDs from extremophilic microorganisms may offer solutions. Expand
Halide-stabilizing residues of haloalkane dehalogenases studied by quantum mechanic calculations and site-directed mutagenesis.
TLDR
The level of stabilization of the transition state structure of S(N)2 reaction and halide ion provided by each of the active site residues in the enzymes DhlA, LinB, and DhaA was quantified by quantum mechanic calculations. Expand
Redesigning dehalogenase access tunnels as a strategy for degrading an anthropogenic substrate.
TLDR
Rhodococcus rhodochrous haloalkane dehalogenase mutants with up to 32-fold higher activity than wild type are obtained toward the toxic, recalcitrant anthropogenic compound 1,2,3-trichloropropane using a new strategy that identifies key residues in access tunnels connecting the buried active site with bulk solvent by rational design and randomized them by directed evolution. Expand
Modification of Activity and Specificity of Haloalkane Dehalogenase from Sphingomonas paucimobilis UT26 by Engineering of Its Entrance Tunnel*
TLDR
The strategy of using site-directed mutagenesis to modify a specific entrance tunnel residue identified by structural and phylogenetic analyses, rather than combinatorial screening, generated a high percentage of viable mutants. Expand
Pathways and mechanisms for product release in the engineered haloalkane dehalogenases explored using classical and random acceleration molecular dynamics simulations.
TLDR
It is proposed that engineering the accessibility of tunnels and the mechanisms of ligand exchange is a powerful strategy for modification of the functional properties of enzymes with buried active sites. Expand
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