Halohydrin dehalogenase (gene HHDH) from Agrobacterium radiobacter AD1

Enzyme Description

Extremophile
No
EC Number

Sequence

Length: 254 amino acids
MSTAIVTNVKHFGGMGSALRLSEAGHTVACHDESFKQKDELEAFAETYPQLKPMSEQEPAELIEAVTSAYGQVDVLVSNDIFAPEFQPIDKYAVEDYRGAVEALQIRPFALVNAVASQMKKRKSGHIIFITSATPFGPWKELSTYTSARAGACTLANALSKELGEYNIPVFAIGPNYLHSEDSPYFYPTEPWKTNPEHVAHVKKVTALQRLGTQKELGELVAFLASGSCDYLTGQVFWLAGGFPMIERWPGMPE
Nevena MilčiΔ‡ et al. (2022) β€” Inhibitory Effect of DMSO on Halohydrin Dehalogenase: Experimental and Computational Insights into the Influence of an Organic Co-solvent on the Structural and Catalytic Properties of a Biocatalyst
Chemistry–A European Journal  Β· doi:10.1002/chem.202201923 β†—  Β· Activity - Classical Stability - deltaHTm Stability - Inactivation Rate Constant Stability - Incubation Stability - Tm Stability - Half-life
73 measurements
Database ID
UniProt: Q93D82 β†—
Sequence Annotation
Explicit - Provided PDB Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli MC1061

Experimental Data (73 measurements)

73 measurements β€” page 3 of 4
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Stability - Inactivation Rate Constant Deactivation constant (in 1/min), determined by residual activity measurements (HPLC) after incubation at 25Β°C Dimethyl Sulfoxide (DMSO) 30% 2.22*10^-5 2.39*10^-5 1/min Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Classical aqueous control (in 1/min) | Enzyme cell-free extracts used for measurements
Stability - Inactivation Rate Constant Deactivation constant (in 1/min), determined by residual activity measurements (HPLC) after incubation at 25Β°C Dimethyl Sulfoxide (DMSO) 50% 2.22*10^-5 1.67*10^-2 1/min Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Classical aqueous control (in 1/min) | Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (52 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 20% 100.0 90.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (24 hours at 25Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 30% 97.0 96.5 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Water-incubated control (in %) | Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (24 hours at 25Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 40% 97.0 67.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Water-incubated control (in %) | Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (24 hours at 25Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 50% 97.0 0.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Water-incubated control (in %) | Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (4 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 5% 100.0 100.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (24 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 5% 100.0 95.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (48 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 5% 100.0 95.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (72 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 5% 100.0 89.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (2 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 20% 100.0 97.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (29 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 20% 100.0 98.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (48 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 30% 100.0 91.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (72 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 20% 100.0 93.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (24 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 30% 100.0 92.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (76 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 30% 100.0 89.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (2 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 40% 100.0 93.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (5 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 40% 100.0 90.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (24 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 40% 100.0 66.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
Stability - Incubation Activity after incubation (48 hours at 50Β°C), measured by HPLC in aqueous phase Dimethyl Sulfoxide (DMSO) 40% 100.0 46.0 % Incubation: 100 mM Tris-SO4 buffer, Assay: 100 mM Tris-SO4 buffer Incubation: 7.5, Assay: 7.5 Incubation: 25Β°C, Assay: 25Β°C 5 mM p-Nitro-2-bromo-1-phenylethanol para-nitro styrene oxide β€” Incubation: 1000 rpm Non-incubated control (in %), assay in aqueous phase | Uncertainty about whether the assay really was in aqueous phase/Enzyme cell-free extracts used for measurements
1 2 3 4

Visualization : Activity β€” Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability β€” Tm

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability β€” Ξ”HTm

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability β€” Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability β€” Half-life

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability β€” Inactivation Rate Constant

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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