Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(lambdaDE3) | Niallia circulans |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant enzyme, hanging drop method, 4°C, equilibration of 2 ml of protein solution, with 25-30 mg/ml protein in 20 mM sodium phosphate buffer, pH 7.0, against an equal volume of 13-20% saturated (NH4)2SO4 in 40 mM Tris-HCl, pH 8.0, X-ray diffraction structure determination and analysis | Niallia circulans |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of a library of circular permutants, generated by random circular permutation of Bcx, random DNase cleavage of the circularized Bcx gene, to introduce new termini in loop regions while linking its native termini directly or via one or two glycines, qualitative analysis, overview. Several permutations place key catalytic residues at or near the new termini with minimal deleterious effects on activity, up to 4fold increased activity. Mutant structure determination and analysis by X-ray diffraction and by NMR spectroscopy. Detailed stability and activity studies on three selected permutants, cpN35G2', cpY94G2', and cpY174G2', overview | Niallia circulans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2,4-dinitrophenyl 2-deoxy-2-fluoro-beta-D-xylobioside | 2,4-dinitrophenolate is released, due to covalent inactivation of the active enzyme, used for active site titrations | Niallia circulans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics of recombinant wild-type and mutant enzymes, overview | Niallia circulans | |
2.4 | - |
2,5-dinitrophenyl-beta-D-xylobioside | recombinant wild-type enzyme, pH 7.2, 22°C | Niallia circulans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Niallia circulans | P09850 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(lambdaDE3) by cation exchange chromatography and gel filtration | Niallia circulans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,5-dinitrophenyl-beta-D-xylobioside + H2O | - |
Niallia circulans | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | Bcx possesses a beta-jellyroll fold that places its N- and C-termini in salt bridge contact. Mutant enzyme structure determination and analysis by X-ray diffraction and by NMR spectroscopy, overview. Overall conformation of Bcx changes very little in response to circular permutation, with effects largely being limited to increased local mobility near the new and the linked old termini and to a decrease in global stability against thermal denaturation | Niallia circulans |
Synonyms | Comment | Organism |
---|---|---|
Bcx | - |
Niallia circulans |
endoxylanase | - |
Niallia circulans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at | Niallia circulans |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
thermal denaturation measurements, circular dichroism, overview | Niallia circulans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
34 | - |
2,5-dinitrophenyl-beta-D-xylobioside | recombinant wild-type enzyme, pH 7.2, 22°C | Niallia circulans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Niallia circulans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
14 | - |
2,5-dinitrophenyl-beta-D-xylobioside | recombinant wild-type enzyme, pH 7.2, 22°C | Niallia circulans |