Literature summary for 2.4.1.10 extracted from

Kanjanatanin, P.; Pichyangkura, R.; Sitthiyotha, T.; Charoenwongpaiboon, T.; Wangpaiboon, K.; Chunsrivirot, S.
Computational design of Bacillus licheniformis RN-01 levansucrase for control of the chain length of levan-type fructooligosaccharides (2019), Int. J. Biol. Macromol., 140, 1239-1248 .

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Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli Top10 cells	Bacillus licheniformis

Protein Variants

Protein Variants	Comment	Organism
E349F	inactive	Bacillus licheniformis
E349F/K372Y	inactive	Bacillus licheniformis
E349W	inactive	Bacillus licheniformis
K372A	the mutant shows reduced activity compared to the wild type enzyme	Bacillus licheniformis
K372Y	the mutant shows reduced activity compared to the wild type enzyme	Bacillus licheniformis
N251W	the mutant shows reduced activity compared to the wild type enzyme	Bacillus licheniformis
N251W/E349F	inactive	Bacillus licheniformis
N251W/E349W/K372Y	inactive	Bacillus licheniformis
N251W/K372Y	the mutant shows reduced activity compared to the wild type enzyme	Bacillus licheniformis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
19.6	-	sucrose	wild type enzyme, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
44.46	-	sucrose	wild type enzyme, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
44.74	-	sucrose	mutant enzyme K372Y, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
77.91	-	sucrose	mutant enzyme K372Y, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
86.69	-	sucrose	mutant enzyme K372A, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
113.13	-	sucrose	mutant enzyme K372A, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
127.96	-	sucrose	mutant enzyme N251W, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
127.96	-	sucrose	wild type enzyme, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
137.62	-	sucrose	mutant enzyme N251W/K372Y, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
176.72	-	sucrose	mutant enzyme K372Y, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
193.4	-	sucrose	mutant enzyme K372A, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
795.22	-	sucrose	mutant enzyme N251W/K372Y, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
820.75	-	sucrose	mutant enzyme N251W, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
978.37	-	sucrose	mutant enzyme N251W, total activity, at pH 6.0 and 50°C	Bacillus licheniformis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
sucrose + [beta-D-fructofuranosyl-(2->6)]n alpha-D-glucopyranoside	Bacillus licheniformis	-	D-glucose + [beta-D-fructofuranosyl-(2->6)]n+1 alpha-D-glucopyranoside	-	?
sucrose + [beta-D-fructofuranosyl-(2->6)]n alpha-D-glucopyranoside	Bacillus licheniformis RN-01	-	D-glucose + [beta-D-fructofuranosyl-(2->6)]n+1 alpha-D-glucopyranoside	-	?

Organism

Organism	UniProt	Comment	Textmining
Bacillus licheniformis	-	-	-
Bacillus licheniformis RN-01	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
Toyopearl DEAE-650M column chromatography	Bacillus licheniformis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	the enzyme also hydrolyzes sucrose to D-glucose and D-fructose	Bacillus licheniformis	?	-	-
additional information	the enzyme also hydrolyzes sucrose to D-glucose and D-fructose	Bacillus licheniformis RN-01	?	-	-
sucrose + sucrose	-	Bacillus licheniformis	D-glucose + 1-kestose + nystose + ?	-	?
sucrose + sucrose	-	Bacillus licheniformis RN-01	D-glucose + 1-kestose + nystose + ?	-	?
sucrose + [beta-D-fructofuranosyl-(2->6)]n alpha-D-glucopyranoside	-	Bacillus licheniformis	D-glucose + [beta-D-fructofuranosyl-(2->6)]n+1 alpha-D-glucopyranoside	-	?
sucrose + [beta-D-fructofuranosyl-(2->6)]n alpha-D-glucopyranoside	-	Bacillus licheniformis RN-01	D-glucose + [beta-D-fructofuranosyl-(2->6)]n+1 alpha-D-glucopyranoside	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
41.8	-	sucrose	mutant enzyme K372A, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
58.24	-	sucrose	mutant enzyme N251W/K372Y, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
81.77	-	sucrose	mutant enzyme N251W, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
99.69	-	sucrose	mutant enzyme K372A, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
144.68	-	sucrose	mutant enzyme K372Y, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
156.86	-	sucrose	mutant enzyme K372A, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
207.11	-	sucrose	wild type enzyme, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
214.01	-	sucrose	mutant enzyme K372Y, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
220.35	-	sucrose	mutant enzyme N251W/K372Y, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
326.62	-	sucrose	mutant enzyme K372Y, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
373.22	-	sucrose	wild type enzyme, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
672.38	-	sucrose	wild type enzyme, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
758.78	-	sucrose	mutant enzyme N251W, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
1118.63	-	sucrose	mutant enzyme N251W, total activity, at pH 6.0 and 50°C	Bacillus licheniformis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.28	-	sucrose	mutant enzyme N251W/K372Y, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
0.42	-	sucrose	mutant enzyme N251W/K372Y, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
0.46	-	sucrose	wild type enzyme, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
0.48	-	sucrose	mutant enzyme K372A, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
0.64	-	sucrose	mutant enzyme N251W, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
0.81	-	sucrose	mutant enzyme K372A, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
0.88	-	sucrose	mutant enzyme K372A, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
0.92	-	sucrose	mutant enzyme N251W, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
1.14	-	sucrose	mutant enzyme N251W, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
1.21	-	sucrose	mutant enzyme K372Y, transfructosylation activity, at pH 6.0 and 50°C	Bacillus licheniformis
3.23	-	sucrose	mutant enzyme K372Y, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis
4.19	-	sucrose	mutant enzyme K372Y, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
8.45	-	sucrose	wild type enzyme, total activity, at pH 6.0 and 50°C	Bacillus licheniformis
10.57	-	sucrose	wild type enzyme, hydrolytic activity, at pH 6.0 and 50°C	Bacillus licheniformis

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Release 2023.1 (January 2023)