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Literature summary for 3.2.1.141 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
Search for more literature for 3.2.1.141

Application

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

Cloned(Commentary)

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

Protein Variants

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

KM Value [mM]

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

Molecular Weight [Da]

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

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus
-
 ATCC 35092
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
maltoheptaose + H2O
-
 Saccharolobus solfataricus ?
-
 ?
maltohexaose + H2O
-
 Saccharolobus solfataricus ?
-
 ?
maltooligosacccharide + H2O
-
 Saccharolobus solfataricus D-glucose
-
 ?
maltooligosyltrehalose + H2O alpha-1,4-glucosidic and alpha-1,1-linked terminal dissaccharides of maltooligosyltrehalose Saccharolobus solfataricus maltooligosaccharide + alpha,alpha'-trehalose trehalose ?
maltopentaose + H2O
-
 Saccharolobus solfataricus ?
-
 ?
maltopentaosyltrehalose + H2O
-
 Saccharolobus solfataricus maltopentaose + alpha,alpha'-trehalose
-
 ?
maltotetraosyltrehalose + H2O
-
 Saccharolobus solfataricus maltotetraose + alpha,alpha'-trehalose
-
 ?
maltotriosyltrehalose + H2O
-
 Saccharolobus solfataricus maltotriose + alpha,alpha'-trehalose
-
 ?

Synonyms

Synonyms Comment Organism
maltooligosyltrehalose trehalohydrolase
-
 Saccharolobus solfataricus
MTHase
-
 Saccharolobus solfataricus

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

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

pH Optimum

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