Literature summary for 3.2.1.156 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.

Search for more literature for 3.2.1.156

Application

Application	Comment	Organism
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)

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

Protein Variants

Protein Variants	Comment	Organism
D263N	mutations at D263 results in decreased F- releasing activity	Halalkalibacterium halodurans
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
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]

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

Organism

Organism	UniProt	Comment	Textmining
Halalkalibacterium halodurans	-	-	-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms	Comment	Organism
Rex	-	Halalkalibacterium halodurans

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	-	Halalkalibacterium halodurans

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Release 2023.1 (January 2023)