Literature summary for 5.1.3.11 extracted from

Kuschel, B.; Claa�en, W.; Mu, W.; Jiang, B.; Stressler, T.; Fischer, L.
Reaction investigation of lactulose-producing cellobiose 2-epimerases under operational relevant conditions (2016), J. Mol. Catal. B, 133, S80-S87 .

No PubMed abstract available

Search for more literature for 5.1.3.11

Application

Application	Comment	Organism
food industry	the enzyme is of interest for the dairy industry due to their ability to convert lactose into epilactose and lactulose, rare disaccharides with prebiotic properties	Caldicellulosiruptor saccharolyticus
food industry	the enzyme is of interest for the dairy industry due to their ability to convert lactose into epilactose and lactulose, rare disaccharides with prebiotic properties	Dictyoglomus turgidum
pharmacology	synthesis of lactulose, which is used as a pharmaceutical against various illnesses, such as chronic constipation	Caldicellulosiruptor saccharolyticus
pharmacology	synthesis of lactulose, which is used as a pharmaceutical against various illnesses, such as chronic constipation	Dictyoglomus turgidum
synthesis	the enzyme from Caldicellulosiruptor saccharolyticus is applied for epilactose and lactulose production in milk at lower temperatures	Caldicellulosiruptor saccharolyticus

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli XL-1	Caldicellulosiruptor saccharolyticus
expression in Escherichia coli XL-1	Dictyoglomus turgidum
recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)	Caldicellulosiruptor saccharolyticus
recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3)	Dictyoglomus turgidum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	Michaelis-Menten kinetics	Caldicellulosiruptor saccharolyticus
additional information	-	additional information	Michaelis-Menten kinetics	Dictyoglomus turgidum
257	-	lactulose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
257	-	lactulose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus
276	-	epilactose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
276	-	epilactose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus
339	-	lactulose	pH 7.5, 50°C	Dictyoglomus turgidum
339	-	lactulose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
381	-	epilactose	pH 7.5, 50°C	Dictyoglomus turgidum
381	-	epilactose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
462	-	lactose	pH 7.5, 50°C	Dictyoglomus turgidum
462	-	lactose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
553	-	lactose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
553	-	lactose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
42000	-	-	Caldicellulosiruptor saccharolyticus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
2 lactulose	Caldicellulosiruptor saccharolyticus	-	lactose + epilactose	-	r
2 lactulose	Dictyoglomus turgidum	-	lactose + epilactose	-	r
2 lactulose	Dictyoglomus turgidum DSM 6724	-	lactose + epilactose	-	r

Organism

Organism	UniProt	Comment	Textmining
Caldicellulosiruptor saccharolyticus	A4XGA6	-	-
Dictyoglomus turgidum	B8DZK4	-	-
Dictyoglomus turgidum DSM 6724	B8DZK4	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Caldicellulosiruptor saccharolyticus
-	Dictyoglomus turgidum
recombinant His-tagged enzyme 151fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage by HRV 3C protease, desalting gel filtration, and another step of nickel affinity chromatography	Dictyoglomus turgidum
recombinant His-tagged enzyme 6.5fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage by HRV 3C protease, desalting gel filtration, and another step of nickel affinity chromatography	Caldicellulosiruptor saccharolyticus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
74.39	-	purified recombinant enzyme, pH 7.5, 50°C, substrate lactose	Caldicellulosiruptor saccharolyticus
90.58	-	purified recombinant enzyme, pH 7.5, 50°C, substrate lactose	Dictyoglomus turgidum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2 lactulose	-	Caldicellulosiruptor saccharolyticus	lactose + epilactose	-	r
2 lactulose	-	Dictyoglomus turgidum	lactose + epilactose	-	r
2 lactulose	-	Dictyoglomus turgidum DSM 6724	lactose + epilactose	-	r
epilactose	lactose production occurs earlier and faster than lactulose production. The lactose content decreases after an initial increase. Simultaneously, the epilactose content decreases rapidly during this first part of the conversion to about 26%. Therefore, 20 min into the conversion of epilactose as a starting substrate, an inversion of the amounts of the two sugars occurr, resulting in about 26% epilactose and 66% lactose	Caldicellulosiruptor saccharolyticus	lactose	-	r
epilactose	lactose production occurs earlier and faster than lactulose production. The lactose content decreases after an initial increase. Simultaneously,the epilactose content decreases rapidly during this first part of the conversion to about 26%. Therefore, 20 min into the conversion of epilactose as a starting substrate, an inversion of the amounts of the two sugars occurrs, resulting in about 26% epilactose and 66% lactose	Dictyoglomus turgidum	lactose	-	r
epilactose	lactose production occurs earlier and faster than lactulose production. The lactose content decreases after an initial increase. Simultaneously,the epilactose content decreases rapidly during this first part of the conversion to about 26%. Therefore, 20 min into the conversion of epilactose as a starting substrate, an inversion of the amounts of the two sugars occurrs, resulting in about 26% epilactose and 66% lactose	Dictyoglomus turgidum DSM 6724	lactose	-	r
epilactose	lactose production occurs earlier and faster than lactulose production. The lactose content decreases after an initial increase. Simultaneously,the epilactose content decreases rapidly during this first part of the conversion to about 26%. Therefore, 20 min into the conversion of epilactose as a starting substrate, an inversion of the amounts of the two sugars occurr, resulting in about 26% epilactose and 66% lactose	Caldicellulosiruptor saccharolyticus	lactulose	-	r
epilactose	lactose production occurs earlier and faster than lactulose production. The lactose content decreases after an initial increase. Simultaneously,the epilactose content decreases rapidly during this first part of the conversion to about 26%. Therefore, 20 min into the conversion of epilactose as a starting substrate, an inversion of the amounts of the two sugars occurrs, resulting in about 26% epilactose and 66% lactose	Dictyoglomus turgidum	lactulose	-	r
epilactose	lactose production occurs earlier and faster than lactulose production. The lactose content decreases after an initial increase. Simultaneously,the epilactose content decreases rapidly during this first part of the conversion to about 26%. Therefore, 20 min into the conversion of epilactose as a starting substrate, an inversion of the amounts of the two sugars occurrs, resulting in about 26% epilactose and 66% lactose	Dictyoglomus turgidum DSM 6724	lactulose	-	r
lactose	the epimerisation and, therefore, the production of epilactose can be seen to occure arlier and faster than the production of lactulose. After reaching a maximal ratio of about 28% after 10 min, the epilactose content decreases and so does the lactose content, while the lactulose content increases over the course of the whole conversion. The conversion reaches an apparent steady state with apparent final sugar ratios of 54.4% lactulose, 14.8% epilactose and 30.8% lactose	Dictyoglomus turgidum	epilactose	-	r
lactose	the epimerisation and, therefore, the production of epilactose can be seen to occure arlier and faster than the production of lactulose. After reaching a maximal ratio of about 28% after 10 min, the epilactose content decreases and so does the lactose content, while the lactulose content increases over the course of the whole conversion. The conversion reaches an apparent steady state with apparent final sugar ratios of 55.0% lactulose, 14.6% epilactose and 30.4% lactose	Caldicellulosiruptor saccharolyticus	epilactose	-	r
lactose	the epimerisation and, therefore, the production of epilactose can be seen to occure arlier and faster than the production of lactulose. After reaching a maximal ratio of about 28% after 10 min, the epilactose content decreases and so does the lactose content, while the lactulose content increases over the course of the whole conversion. The conversion reaches an apparent steady state with apparent final sugar ratios of 54.4% lactulose, 14.8% epilactose and 30.8% lactose	Dictyoglomus turgidum DSM 6724	epilactose	-	r
lactose	the epimerisation and, therefore, the production of epilactose can be seen to occure arlier and faster than the production of lactulose. After reaching a maximal ratio of about 28% after 10 min, the epilactose content decreases and so does the lactose content, while the lactulose content increases over the course of the whole conversion. The conversion reaches an apparent steady state with apparent final sugar ratios of 54.4% lactulose, 14.8% epilactose and 30.8% lactose	Dictyoglomus turgidum	lactulose	-	r
lactose	the epimerisation and, therefore, the production of epilactose can be seen to occure earlier and faster than the production of lactulose. After reaching a maximal ratio of about 28% after 10 min, the epilactose content decreases and so does the lactose content, while the lactulose content increases over the course of the whole conversion. The conversion reaches an apparent steady state with apparent final sugar ratios of 55.0% lactulose, 14.6% epilactose and 30.4% lactose	Caldicellulosiruptor saccharolyticus	lactulose	-	r
lactose	the epimerisation and, therefore, the production of epilactose can be seen to occure arlier and faster than the production of lactulose. After reaching a maximal ratio of about 28% after 10 min, the epilactose content decreases and so does the lactose content, while the lactulose content increases over the course of the whole conversion. The conversion reaches an apparent steady state with apparent final sugar ratios of 54.4% lactulose, 14.8% epilactose and 30.8% lactose	Dictyoglomus turgidum DSM 6724	lactulose	-	r

Subunits

Subunits	Comment	Organism
?	x * 42000, recombinant detagged enzyme, SDS-PAGE, x * 47300, about sequence calculation	Caldicellulosiruptor saccharolyticus
?	x * 42000, recombinant detagged enzyme, SDS-PAGE, x * 47500, about sequence calculation	Dictyoglomus turgidum

Synonyms

Synonyms	Comment	Organism
cellobiose 2-epimerase	-	Caldicellulosiruptor saccharolyticus
cellobiose 2-epimerase	-	Dictyoglomus turgidum
CsCE	-	Caldicellulosiruptor saccharolyticus
DsCE	-	Dictyoglomus turgidum
DtCE	-	Dictyoglomus turgidum

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	-	Caldicellulosiruptor saccharolyticus
50	-	-	Dictyoglomus turgidum
70	-	-	Dictyoglomus turgidum
80	-	-	Caldicellulosiruptor saccharolyticus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.54	-	lactulose	pH 7.5, 50°C	Dictyoglomus turgidum
0.54	-	lactulose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
0.88	-	lactulose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
0.88	-	lactulose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus
101	-	epilactose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
101	-	epilactose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus
125	-	epilactose	pH 7.5, 50°C	Dictyoglomus turgidum
125	-	epilactose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
237	-	lactose	pH 7.5, 50°C	Dictyoglomus turgidum
237	-	lactose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
303	-	lactose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
303	-	lactose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	-	Dictyoglomus turgidum
7.5	-	-	Caldicellulosiruptor saccharolyticus
7.5	-	-	Dictyoglomus turgidum

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.0016	-	lactulose	pH 7.5, 50°C	Dictyoglomus turgidum
0.0016	-	lactulose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
0.0034	-	lactulose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
0.0034	-	lactulose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus
0.33	-	epilactose	pH 7.5, 50°C	Dictyoglomus turgidum
0.33	-	epilactose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
0.37	-	epilactose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
0.37	-	epilactose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus
0.51	-	lactose	pH 7.5, 50°C	Dictyoglomus turgidum
0.51	-	lactose	pH 7.5, 50°C, recombinant enzyme	Dictyoglomus turgidum
0.55	-	lactose	pH 7.5, 50°C	Caldicellulosiruptor saccharolyticus
0.55	-	lactose	pH 7.5, 50°C, recombinant enzyme	Caldicellulosiruptor saccharolyticus

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Release 2023.1 (January 2023)