Literature summary for 2.4.1.10 extracted from

Rairakhwada, D.; Seo, J.W.; Seo, M.Y.; Kwon, O.; Rhee, S.K.; Kim, C.H.
Gene cloning, characterization, and heterologous expression of levansucrase from Bacillus amyloliquefaciens (2010), J. Ind. Microbiol. Biotechnol., 37, 195-204.

Cloned(Commentary)

Cloned (Comment)	Organism
gene sacB, cloning in Escherichia coli strain DH5alpha, expression in Escherichia coli strain BL21(DE3), and high xylose-inducible expression of the His6-tagged enzyme in Bacillus megaterium, where the recombinant enzyme is secreted to the medium, method optimization by response-surface methodology, overview. The enhanced secretion of His6-tagged protein is an advantage for purification via the C-terminal His6 tag, which leads to the purification of intact levansucrase, and a 21fold higher compared to the untagged enzyme	Bacillus amyloliquefaciens

Cloned (Comment)

Organism

gene sacB, cloning in Escherichia coli strain DH5alpha, expression in Escherichia coli strain BL21(DE3), and high xylose-inducible expression of the His6-tagged enzyme in Bacillus megaterium, where the recombinant enzyme is secreted to the medium, method optimization by response-surface methodology, overview. The enhanced secretion of His6-tagged protein is an advantage for purification via the C-terminal His6 tag, which leads to the purification of intact levansucrase, and a 21fold higher compared to the untagged enzyme

Bacillus amyloliquefaciens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetics, overview	Bacillus amyloliquefaciens
47.81	-	D-glucose	pH 6.0, 30°C, recombinant enzyme	Bacillus amyloliquefaciens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Bacillus amyloliquefaciens	5829	-
extracellular	secretion of the enzyme	Bacillus amyloliquefaciens	-	-

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
49000	-	x * 49000, recombinant enzyme expressed in Escherichia coli, SDS-PAGE	Bacillus amyloliquefaciens

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Bacillus amyloliquefaciens	-	type 1, gene sacB	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged enzyme 4fold by nickel affinity chromatography	Bacillus amyloliquefaciens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
10.8	-	native enzyme from strain type 1, culture supernatant	Bacillus amyloliquefaciens
37.32	-	purified recombinant His-tagged enzyme from Escherichia coli, release of glucose	Bacillus amyloliquefaciens
669.6	-	recombinant His6-tagged enzyme from Bacillus megaterium, culture supernatant	Bacillus amyloliquefaciens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2 sucrose	-	Bacillus amyloliquefaciens	D-glucose + beta-D-fructofuranosyl-(2,6)-beta-D-fructofuranosyl-(2,1)-alpha-D-glucopyranoside	-	?
sucrose + beta-D-fructofuranosyl-[(2,6)beta-D-fructofuranosyl-]n alpha-D-glucopyranoside	-	Bacillus amyloliquefaciens	D-glucose + beta-D-fructofuranosyl-[(2,6)beta-D-fructofuranosyl-]n+1 alpha-D-glucopyranoside	-	?

Subunits

Subunits	Comment	Organism
?	x * 49000, recombinant enzyme expressed in Escherichia coli, SDS-PAGE	Bacillus amyloliquefaciens

Synonyms

Synonyms	Comment	Organism
SacB	-	Bacillus amyloliquefaciens
type 1 levansucrase	-	Bacillus amyloliquefaciens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
4	-	levan formation	Bacillus amyloliquefaciens
30	-	D-glucose production	Bacillus amyloliquefaciens

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
4	60	-	Bacillus amyloliquefaciens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6	-	D-glucose production, recombinant enzyme	Bacillus amyloliquefaciens
8	-	levan formation, recombinant enzyme	Bacillus amyloliquefaciens

pH Range

pH Minimum	pH Maximum	Comment	Organism
4	9	pH activity profile, overview	Bacillus amyloliquefaciens