Literature summary for 2.4.1.10 extracted from

Li, S.Y.; Chen, M.; Li, G.; Yan, Y.L.; Yu, H.Y.; Zhan, Y.H.; Peng, Z.X.; Wang, J.; Lin, M.
Amino acid substitutions of His296 alter the catalytic properties of Zymomonas mobilis 10232 levansucrase (2008), Acta Biochim. Pol., 55, 201-206.

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Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli JM109	Zymomonas mobilis

Protein Variants

Protein Variants	Comment	Organism
H296C	site-directed mutagenesis, no detectable activities	Zymomonas mobilis
H296E	site-directed mutagenesis, no detectable activities	Zymomonas mobilis
H296K	site-directed mutagenesis, minimal hydrolysis and transfructosylation activities, transfructosylation activity is reduced to 4.6% of the wild-type	Zymomonas mobilis
H296L	site-directed mutagenesis, minimal hydrolysis and transfructosylation activities, transfructosylation activity is reduced to 1.0% of the wild-type, mediates the accumulation of low amounts of trisaccharide, but did not produce pentasaccharide	Zymomonas mobilis
H296Q	site-directed mutagenesis, minimal hydrolysis and transfructosylation activities, transfructosylation activity is reduced to 2.5% of the wild-type	Zymomonas mobilis
H296R	site-directed mutagenesis, retains partial hydrolysis and transfructosylation activities, transfructosylation activity is reduced to 23.1% of the wild-type	Zymomonas mobilis
H296S	site-directed mutagenesis, no detectable activities	Zymomonas mobilis
H296W	site-directed mutagenesis, retains partial hydrolysis and transfructosylation activities, transfructosylation activity is reduced to 19.8% of the wild-type	Zymomonas mobilis
H296Y	site-directed mutagenesis, no detectable activities	Zymomonas mobilis
additional information	The His296 mutation influences the substrate recognizing and binding, and then has an effect on sucrose hydrolysis and fructosyl-enzyme intermediate formation. Its mutations weakens the transfructosylation reaction and changes product specificities	Zymomonas mobilis

Organism

Organism	UniProt	Comment	Textmining
Zymomonas mobilis	-	10232 (sacB), expression in Escherichia coli JM109, Zymomonas mobilis levansucrase His296, located in homologous region IV, is equivalent to Bacillus subtilis SacB Arg331 and Gluconacetobacter diazotrophicus LsdA His419	-

Purification (Commentary)

Purification (Comment)	Organism
Ni-NTA column	Zymomonas mobilis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
sucrose + sucrose	is assayed in a standard reaction containing 1 ml 10% sucrose and 1 ml enzyme (0.01-0.05mg/ml) incubated at 15°C	Zymomonas mobilis	D-glucose + levan	-	?

Synonyms

Synonyms	Comment	Organism
levansucrase	-	Zymomonas mobilis

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Release 2023.1 (January 2023)