Literature summary extracted from

  • Fang, T.Y.; Tseng, W.C.; Shih, T.Y.; Wang, M.Y.
    Identification of the essential catalytic residues and selectivity-related residues of maltooligosyltrehalose trehalohydrolase from the thermophilic archaeon Sulfolobus solfataricus ATCC 35092 (2008), J. Agric. Food Chem., 56, 5628-5633.
    View publication on PubMed

Application

EC Number Application Comment Organism
3.2.1.141 additional information trehalose is gaining applications as sweetener component, preservative or stabilizer of food, cosmetics, vaccines, medicines, cells, and organs Saccharolobus solfataricus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.141 Escherichia coli Rosetta (DE3) with mutated vector pET-15b-deltaH-treZ, Terrific broth, 37°C Saccharolobus solfataricus

Engineering

EC Number Protein Variants Comment Organism
3.2.1.141 W218A mutation decreases selectivity ratio glucose/trehalose formation Saccharolobus solfataricus
3.2.1.141 D255A 0.15% of wild-type activity Saccharolobus solfataricus
3.2.1.141 E286A 0.09% of wild-type activity Saccharolobus solfataricus
3.2.1.141 D380A 0.01% of wild-type activity Saccharolobus solfataricus
3.2.1.141 A259S mutation increases selectivity ratio glucose/trehalose formation Saccharolobus solfataricus
3.2.1.141 Y328F mutation increases selectivity ratio glucose/trehalose formation Saccharolobus solfataricus
3.2.1.141 F355Y mutation increases selectivity ratio glucose/trehalose formation Saccharolobus solfataricus
3.2.1.141 R356K mutation increases selectivity ratio glucose/trehalose formation Saccharolobus solfataricus
3.2.1.141 W218F mutation decreases selectivity ratio glucose/trehalose formation Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.141 5.26
-
maltopentaosyltrehalose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 8.9
-
maltotriosyltrehalose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 2.19
-
maltoheptaose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 2.19
-
maltoheptaose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 10.9
-
maltotriosyltrehalose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 2.16
-
maltohexaose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 2.63
-
maltoheptaose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 5.08
-
maltopentaose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 5.89
-
maltopentaosyltrehalose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 13.6
-
maltotriosyltrehalose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 3.01
-
maltohexaose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 5.37
-
maltopentaose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 8.55
-
maltotriosyltrehalose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 2.81
-
maltoheptaose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 3.42
-
maltohexaose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 7.15
-
maltopentaosyltrehalose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5.63
-
maltohexaose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 23.2
-
maltohexaose mutant enzyme W218F, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 20.5
-
maltohexaose mutant enzyme W218A, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 2.88
-
maltoheptaose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 6.79
-
maltotetraosyltrehalose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 6.79
-
maltotetraosyltrehalose mutant enzyme W218F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5.03
-
maltotetraosyltrehalose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 7.22
-
maltotriosyltrehalose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 6.94
-
maltotetraosyltrehalose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5.82
-
maltopentaose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 5.66
-
maltotetraosyltrehalose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 6.73
-
maltotetraosyltrehalose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5.81
-
maltopentaosyltrehalose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 7.91
-
maltotetraosyltrehalose mutant enzyme W218A, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5.75
-
maltopentaosyltrehalose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5.86
-
maltopentaose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 4.69
-
maltohexaose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 7.74
-
maltopentaose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.141 129000
-
mass of wild-type and mutant enzymes, BioSep-SEC-S4000 gel filtration column Saccharolobus solfataricus

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.141 Saccharolobus solfataricus
-
ATCC 35092
-
5.4.99.15 Saccharolobus solfataricus
-
strain ATCC 35092
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.141 Q-Sepharose anion exchange column chromatography Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.141 0.1
-
D380A MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 5.31
-
W218A MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 52
-
W218F MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 814
-
wild-type MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 1.23
-
D255A MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 0.74
-
E286A MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 492
-
Y328F MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 472
-
R356K MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 597
-
F355Y MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 787
-
A259S MTHase, hydrolysis of maltotetraosyltrehalose at 60°C in 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.141 maltotriosyltrehalose + H2O
-
Saccharolobus solfataricus maltotriose + alpha,alpha'-trehalose
-
?
3.2.1.141 maltotetraosyltrehalose + H2O
-
Saccharolobus solfataricus maltotetraose + alpha,alpha'-trehalose
-
?
3.2.1.141 maltopentaose + H2O
-
Saccharolobus solfataricus ?
-
?
3.2.1.141 maltohexaose + H2O
-
Saccharolobus solfataricus ?
-
?
3.2.1.141 maltoheptaose + H2O
-
Saccharolobus solfataricus ?
-
?
3.2.1.141 maltopentaosyltrehalose + H2O
-
Saccharolobus solfataricus maltopentaose + alpha,alpha'-trehalose
-
?
3.2.1.141 4'-O-(1,4-alpha-D-glucosyl)n-alpha,alpha'-trehalose + H2O with isoamylase and maltooligosyltrehalose synthase Saccharolobus solfataricus (1,4-alpha-D-glucosyl)n + alpha,alpha'-trehalose
-
?
5.4.99.15 soluble starch
-
Saccharolobus solfataricus trehalose
-
?
3.2.1.141 maltooligosacccharide + H2O
-
Saccharolobus solfataricus D-glucose
-
?
3.2.1.141 maltooligosyltrehalose + H2O alpha-1,4-glucosidic and alpha-1,1-linked terminal dissaccharides of maltooligosyltrehalose Saccharolobus solfataricus maltooligosaccharide + alpha,alpha'-trehalose trehalose ?

Synonyms

EC Number Synonyms Comment Organism
5.4.99.15 MTSase
-
Saccharolobus solfataricus
3.2.1.141 maltooligosyltrehalose trehalohydrolase
-
Saccharolobus solfataricus
3.2.1.141 MTHase
-
Saccharolobus solfataricus
5.4.99.15 maltooligosyltrehalose synthase
-
Saccharolobus solfataricus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.141 85
-
wild-type, W218A, W218F, A259S, Y328F, F355Y, and R356K MTHases Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.141 80
-
all enzyme activity retained after 2 hours at 80°C, wild-type, W218A, W218F, A259S, Y328F, F355Y, and R356K MTHases Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.141 16
-
maltotetraosyltrehalose mutant enzyme W218A, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 13.5
-
maltohexaose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 760
-
maltopentaosyltrehalose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5
-
maltoheptaose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 17.7
-
maltopentaose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 12.6
-
maltohexaose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 15.5
-
maltopentaose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 0.36
-
maltohexaose mutant enzyme W218A, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 17.4
-
maltopentaose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 14.4
-
maltopentaose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5 Saccharolobus solfataricus
3.2.1.141 0.54
-
maltohexaose mutant enzyme W218F, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 19.1
-
maltopentaose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 9.5
-
maltohexaose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 721
-
maltotetraosyltrehalose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 1030
-
maltotriosyltrehalose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 18.2
-
maltohexaose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 634
-
maltotriosyltrehalose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 675
-
maltopentaosyltrehalose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 627
-
maltotetraosyltrehalose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 654
-
maltotetraosyltrehalose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 1002
-
maltotriosyltrehalose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 5.49
-
maltoheptaose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 572
-
maltotriosyltrehalose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 58.4
-
maltotetraosyltrehalose mutant enzyme W218F, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 6.01
-
maltoheptaose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 741
-
maltopentaosyltrehalose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 734
-
maltotriosyltrehalose mutant enzyme F355Y, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 1192
-
maltotetraosyltrehalose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 1106
-
maltotetraosyltrehalose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 1226
-
maltopentaosyltrehalose wild-type enzyme, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 1282
-
maltopentaosyltrehalose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, trehalose formation Saccharolobus solfataricus
3.2.1.141 8.23
-
maltohexaose mutant enzyme Y328F, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 6.49
-
maltoheptaose mutant enzyme A259S, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus
3.2.1.141 6.49
-
maltoheptaose mutant enzyme R356K, 60°C, 50 mM citrate phosphate buffer, pH 5, glucose formation Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.141 5
-
wild-type, W218A, W218F, A259S, Y328F, F355Y, and R356K MTHases Saccharolobus solfataricus