Literature summary for 2.4.1.4 extracted from

Verges, A.; Barbe, S.; Cambon, E.; Moulis, C.; Tranier, S.; Remaud-Simeon, M.; Andre, I.
Engineering of anp efficient mutant of Neisseria polysaccharea amylosucrase for the synthesis of controlled size maltooligosaccharides (2017), Carbohydr. Polym., 173, 403-411 .

Search for more literature for 2.4.1.4

Application

Application	Comment	Organism
synthesis	mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A only produces soluble oligosaccharides as no insoluble high molecular weight amylose is observed. The mutant enzyme is an attractive enzymatic tool that could offer interesting opportunities for the design of amylodextrins with controlled size	Neisseria polysaccharea

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Neisseria polysaccharea

Crystallization (Commentary)

Crystallization (Comment)	Organism
the mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A is crystallized by hanging drop vapor diffusion method at 12°C	Neisseria polysaccharea

Protein Variants

Protein Variants	Comment	Organism
R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A	mutant enzyme that shows a 6fold enhanced activity toward sucrose compared to the wild-type enzyme. Only soluble maltooligosaccharide products of controlled size chains (2 < DP < 21) with a narrow polydispersity are observed. This variant, containing 9 mutations in the active site, is characterized at both biochemical and structural levels. Its X-ray structure is determined and further investigated by molecular dynamics to understand the molecular origins of its higher activity on sucrose and higher production of small maltooligosaccharides, with a totally abolished insoluble glucan synthesis	Neisseria polysaccharea

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
50.2	-	sucrose	30°C, pH 7.0, wild-type enzyme	Neisseria polysaccharea
128.3	-	sucrose	30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A	Neisseria polysaccharea

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
sucrose + [(1->4)-alpha-D-glucosyl]n	Neisseria polysaccharea	the enzyme catalyzes the synthesis of alpha-1,4 glucans from sucrose. The product profile is quite polydisperse, ranging from soluble chains called maltooligosaccharides to high-molecular weight insoluble amylose	D-fructose + [(1->4)-alpha-D-glucosyl]n+1	-	?

Organism

Organism	UniProt	Comment	Textmining
Neisseria polysaccharea	Q9ZEU2	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Neisseria polysaccharea

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
sucrose + [(1->4)-alpha-D-glucosyl]n	the enzyme catalyzes the synthesis of alpha-1,4 glucans from sucrose. The product profile is quite polydisperse, ranging from soluble chains called maltooligosaccharides to high-molecular weight insoluble amylose	Neisseria polysaccharea	D-fructose + [(1->4)-alpha-D-glucosyl]n+1	-	?
sucrose + [(1->4)-alpha-D-glucosyl]n	mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A only produces soluble oligosaccharides as no insoluble high molecular weight amylose is observed	Neisseria polysaccharea	D-fructose + [(1->4)-alpha-D-glucosyl]n+1	-	?

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
46.1	-	Tm-value, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A	Neisseria polysaccharea
49	-	Tm-value, wild-type enzyme	Neisseria polysaccharea

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
1.3	-	sucrose	30°C, pH 7.0, wild-type enzyme	Neisseria polysaccharea
1.61	-	sucrose	30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A	Neisseria polysaccharea

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.013	-	sucrose	30°C, pH 7.0, mutant enzyme R226K/I228V/A289I/F290Y/E300I/V331T/Q437S/N439D/C445A	Neisseria polysaccharea
0.026	-	sucrose	30°C, pH 7.0, wild-type enzyme	Neisseria polysaccharea

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Release 2023.1 (January 2023)