A5010613040- Microbial bioremediation and biosurfactants
- Enzyme Structure and Function
- Protein Structure and Dynamics
- Enzyme Catalysis and Immobilization
- Microbial Metabolic Engineering and Bioproduction
- Microbial metabolism and enzyme function
- Distributed and Parallel Computing Systems
- Chemical Reactions and Isotopes
- Biofuel production and bioconversion
- Hemoglobin structure and function
- Electrochemical sensors and biosensors
- Microbial Metabolites in Food Biotechnology
- Steroid Chemistry and Biochemistry
- Education, Psychology, and Social Research
- Anaerobic Digestion and Biogas Production
- Computational Drug Discovery Methods
- Surface Chemistry and Catalysis
- Biochemical and Molecular Research
- Distributed systems and fault tolerance
- Biotin and Related Studies
- Enzyme-mediated dye degradation
- Environmental remediation with nanomaterials
- Diet, Metabolism, and Disease
- Ionic liquids properties and applications
- bioluminescence and chemiluminescence research
RECETOX
2010-2019
Masaryk University
2008-2019
One of the major barriers to use enzymes in industrial biotechnology is their insufficient stability under processing conditions. The organic solvent systems instead aqueous media for enzymatic reactions offers numerous advantages, such as increased solubility hydrophobic substrates or suppression water-dependent side reactions. For example, reverse hydrolysis that form esters from acids and alcohols become thermodynamically favorable. However, solvents often inactivate enzymes. Industry...
Mutations targeting as few four residues lining the access tunnel extended half-life of an enzyme in 40 % dimethyl sulfoxide from minutes to weeks and increased its melting temperature by 19 °C. Protein crystallography molecular dynamics revealed that residue packing is a key determinant protein stability active-site accessibility for cosolvent molecules (red dots).
In the loop: Engineering of surface loop in haloalkane dehalogenases affects their enantiodiscrimination behavior. The temperature dependence enantioselectivity (lnE versus 1/T) β-bromoalkanes by is reversed (red data points) deletion loop; selectivity switches back when an additional single-point mutation made. This behavior not observed for α-bromoesters. Detailed facts importance to specialist readers are published as "Supporting Information". Such documents peer-reviewed, but copy-edited...
An enzyme's substrate specificity is one of its most important characteristics. The quantitative comparison broad-specificity enzymes requires the selection a homogenous set substrates for experimental testing, determination substrate-specificity data and analysis using multivariate statistics. We describe systematic specificities nine wild-type four engineered haloalkane dehalogenases. were characterized experimentally 30 selected statistical design from nearly 200 halogenated compounds....
Abstract A variant of the haloalkane dehalogenase DhaA with greatly enhanced stability and tolerance organic solvents but reduced activity was created by mutating four residues in access tunnel. To create a stabilised enzyme superior catalytic activity, two originally modified were randomised. The resulting mutant F 176 G exhibited 32‐ 10‐times towards 1,2‐dibromoethane buffer 40 % DMSO, respectively, upon retaining high stability. Structural molecular dynamics analyses demonstrated that new...
A haloalkane dehalogenase, DpcA, from Psychrobacter cryohalolentis K5, representing a novel psychrophilic member of the dehalogenase family, was identified and biochemically characterized. DpcA exhibited unique temperature profile with exceptionally high activities at low temperatures. The properties make this enzyme promising for various environmental applications.
This study focuses on two representatives of experimentally uncharacterized haloalkane dehalogenases from the subfamily HLD-III. We report biochemical characterization expression products dehalogenase genes drbA Rhodopirellula baltica SH1 and dmbC Mycobacterium bovis 5033/66. The DrbA DmbC enzymes show highly oligomeric structures very low activities with typical substrates dehalogenases.
ABSTRACT We report the biochemical characterization of a novel haloalkane dehalogenase, DatA, isolated from plant pathogen Agrobacterium tumefaciens C58. DatA possesses peculiar pair halide-stabilizing residues, Asn-Tyr, which have not been reported to play this role in other known dehalogenases. has number unique characteristics, including substrate-dependent and cooperative kinetics, dimeric structure, excellent enantioselectivity toward racemic mixtures chiral brominated alkanes esters.
The crystal structure of the novel haloalkane dehalogenase DbeA from Bradyrhizobium elkanii USDA94 revealed presence two chloride ions buried in protein interior. first halide-binding site is involved substrate binding and present all structurally characterized dehalogenases. second unique to DbeA. To elucidate role enzyme functionality, a two-point mutant lacking this was constructed characterized. These substitutions resulted shift substrate-specificity class were accompanied by decrease...
Mutationen von vier Resten entlang des Zugangstunnels verlängerten die Halbwertzeit eines Enzyms in 40 % Dimethylsulfoxid Minuten zu Wochen und erhöhten Schmelztemperatur um 19 °C. Proteinkristallographie Moleküldynamik zeigen, dass Packung der Tunnelreste entscheidend für Proteinstabilität Zugänglichkeit aktiven Zentrums Cosolvens-Moleküle (rote Punkte) ist. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer...
Gezielte Mutationen der Oberflächenschleife von Halogenalkan-Dehalogenasen haben Auswirkungen auf die Enantiodiskriminierung. Die Temperaturabhängigkeit Enantioselektivität (siehe lnE-1/T-Kurve) β-Bromalkanen kehrt sich bei einer Deletion um (rote Datenpunkte), was Einführung zusätzlichen Einzelpunktmutation wieder rückgängig gemacht wird; α-Bromestern findet keine Änderung statt. Detailed facts of importance to specialist readers are published as "Supporting Information". Such documents...
The haloalkane dehalogenase DhaA from Rhodococcus rhodochrous NCIMB 13064 is a bacterial enzyme that shows catalytic activity for the hydrolytic degradation of highly toxic industrial pollutant 1,2,3-trichloropropane (TCP). Mutagenesis focused on access tunnels produced protein variants with significantly improved towards TCP. Three mutants named DhaA04 (C176Y), DhaA14 (I135F) and DhaA15 (C176Y + I135F) were constructed in order to study functional relevance connecting buried active site...
The enzyme DhaA from Rhodococcus rhodochrous NCIMB 13064 belongs to the haloalkane dehalogenases, which catalyze hydrolysis of haloalkanes corresponding alcohols. dehalogenase and its variants can be used detoxify industrial pollutant 1,2,3-trichloropropane (TCP). Three mutants named DhaA04, DhaA14 DhaA15 were constructed in order study importance tunnels connecting buried active site with surrounding solvent enzymatic activity. All protein crystallized using sitting-drop vapour-diffusion...