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Literature summary extracted from

  • Honda, Y.; Fushinobu, S.; Hidaka, M.; Wakagi, T.; Shoun, H.; Taniguchi, H.; Kitaoka, M.
    Alternative strategy for converting an inverting glycoside hydrolase into a glycosynthase (2008), Glycobiology, 18, 325-330.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.2.1.156 biotechnology a novel method for producing a glycosynthase from an inverting glycoside hydrolase by mutating a residue that holds the nucleophilic water molecule (Y198) with the general base residue while keeping the general base residue intact Halalkalibacterium halodurans

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.156 expressed as a His-tagged fusion protein in Escherichia coli Halalkalibacterium halodurans

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.156 D263N mutations at D263 results in decreased F- releasing activity Halalkalibacterium halodurans
3.2.1.156 Y198F a mutation in the tyrosine residue changing it into phenylalanine causes a drastic decrease in the hydrolytic activity and a small increase in F- releasing activity from alpha-xylobiosyl fluoride in the presence of xylose. As a result of the high F- releasing activity and low hydrolytic activity, Y198F of Rex accumulates a large amount of product during the glycosynthase reaction. Km: 14.8 (xylose), kcat: 11.9/sec (xylose) Halalkalibacterium halodurans
3.2.1.156 Y198F/D263N Y198F/D263N showed 20 times less F- releasing activity than Y198F, which is comparable to that of the corresponding single mutant, D263N. Km: 6.0 (xylose), kcat: 0.42/sec (xylose) Halalkalibacterium halodurans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.156 6
-
 D-xylose mutant Y198F/D263N Halalkalibacterium halodurans
3.2.1.156 6.6
-
 D-xylose
-
 Halalkalibacterium halodurans
3.2.1.156 14.8
-
 D-xylose mutant Y198F Halalkalibacterium halodurans

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.156 Halalkalibacterium halodurans
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.156 by using a Ni-NTA agarose column Halalkalibacterium halodurans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.156 alpha-xylobiosyl fluoride + D-xylose
-
 Halalkalibacterium halodurans alpha-xylobiose + HF + D-xylose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.156 Rex
-
 Halalkalibacterium halodurans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.156 30
-
 assay at Halalkalibacterium halodurans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.156 0.42
-
 D-xylose mutant Y198F/D263N Halalkalibacterium halodurans
3.2.1.156 5.04
-
 D-xylose
-
 Halalkalibacterium halodurans
3.2.1.156 11.9
-
 D-xylose mutant Y198F Halalkalibacterium halodurans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.156 7
-
-
 Halalkalibacterium halodurans