Literature summary for 3.2.1.8 extracted from

Reitinger, S.; Yu, Y.; Wicki, J.; Ludwiczek, M.; DAngelo, I.; Baturin, S.; Okon, M.; Strynadka, N.C.; Lutz, S.; Withers, S.G.; McIntosh, L.P.
Circular permutation of Bacillus circulans xylanase: a kinetic and structural study (2010), Biochemistry, 49, 2464-2474.

Search for more literature for 3.2.1.8

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(lambdaDE3)	Niallia circulans

Crystallization (Commentary)

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

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

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 [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

Organism	UniProt	Comment	Textmining
Niallia circulans	P09850	-	-

Purification (Commentary)

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2,5-dinitrophenyl-beta-D-xylobioside + H2O	-	Niallia circulans	?	-	?

Subunits

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

Synonyms	Comment	Organism
Bcx	-	Niallia circulans
endoxylanase	-	Niallia circulans

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at	Niallia circulans

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
additional information	-	thermal denaturation measurements, circular dichroism, overview	Niallia circulans

Turnover Number [1/s]

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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Niallia circulans

kcat/KM [mM/s]

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

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Release 2023.1 (January 2023)