Literature summary extracted from

  • Seo, J.S.; An, J.H.; Cheong, J.J.; Choi, Y.D.; Kim, C.H.
    Bifunctional recombinant fusion enzyme between maltooligosyltrehalose synthase and maltooligosyltrehalose trehalohydrolase of thermophilic microorganism Metallosphaera hakonensis (2008), J. Microbiol. Biotechnol., 18, 1544-1549.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.141 Escherichia coli BL21(DE3)pLysS, LB medium, vector pRSET-B Metallosphaera hakonensis
5.4.99.15 expressed in Escherichia coli BL21(DE3)pLysS cells as fusion enzyme with maltooligosyltrehalose trehalohydrolase Metallosphaera hakonensis

Engineering

EC Number Protein Variants Comment Organism
3.2.1.141 additional information bifunctional fusion enzyme (MhMTSH) of maltooligosyltrehalose synthase (MhMTS) and malooligosyltrehalose trehalohydrolase (MhMTH) Metallosphaera hakonensis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.141 150000
-
SDS-PAGE, in accordance with the calculated value from the amino acid sequence, fusion enzyme including the hexahistidine domain from the vector Metallosphaera hakonensis
5.4.99.15 150000
-
MTSH fusion enzyme Metallosphaera hakonensis

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.141 Metallosphaera hakonensis
-
grown on JCM165 medium at 70°C
-
5.4.99.15 Metallosphaera hakonensis
-
strain JCM8857
-
5.4.99.15 Metallosphaera hakonensis JCM 8857
-
strain JCM8857
-
3.2.1.141 Metallosphaera hakonensis JMC8857
-
grown on JCM165 medium at 70°C
-

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 additional information
-
5 mM maltooligosaccharides, 100mM sodium phosphate buffer, pH 5, 70°C, 30 min, with substrate soluble starch 24 h Metallosphaera hakonensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.141 maltotriosyltrehalose + H2O sequential hydrolysis by the bifunctional fusion enzyme of maltooligosyltrehalose synthase (MhMTS) and malooligosyltrehalose trehalohydrolase (MhMTH), synthase converts alpha-1,4-glycosidic linkage into alpha-1,1 linkage to produce maltotriosyltrehalose, which is the substrate for the MTH to produce trehalose Metallosphaera hakonensis maltotriose + alpha,alpha'-trehalose
-
?
3.2.1.141 maltotriosyltrehalose + H2O sequential hydrolysis by the bifunctional fusion enzyme of maltooligosyltrehalose synthase (MhMTS) and malooligosyltrehalose trehalohydrolase (MhMTH), synthase converts alpha-1,4-glycosidic linkage into alpha-1,1 linkage to produce maltotriosyltrehalose, which is the substrate for the MTH to produce trehalose Metallosphaera hakonensis JCM 8857 maltotriose + alpha,alpha'-trehalose
-
?
3.2.1.141 maltotriosyltrehalose + H2O sequential hydrolysis by the bifunctional fusion enzyme of maltooligosyltrehalose synthase (MhMTS) and malooligosyltrehalose trehalohydrolase (MhMTH), synthase converts alpha-1,4-glycosidic linkage into alpha-1,1 linkage to produce maltotriosyltrehalose, which is the substrate for the MTH to produce trehalose Metallosphaera hakonensis JMC8857 maltotriose + alpha,alpha'-trehalose
-
?
5.4.99.15 maltohexaose
-
Metallosphaera hakonensis alpha-maltotetraosyl trehalose
-
?
5.4.99.15 maltohexaose
-
Metallosphaera hakonensis JCM 8857 alpha-maltotetraosyl trehalose
-
?
5.4.99.15 maltohexaose
-
Metallosphaera hakonensis JMC8857 alpha-maltotetraosyl trehalose
-
?
3.2.1.141 4'-O-(1,4-alpha-D-glucosyl)n-alpha,alpha'-trehalose + H2O hydrolysis to disaccharides after 24 h Metallosphaera hakonensis (1,4-alpha-D-glucosyl)n + alpha,alpha'-trehalose
-
?
3.2.1.141 4'-O-(1,4-alpha-D-glucosyl)n-alpha,alpha'-trehalose + H2O hydrolysis to disaccharides after 24 h Metallosphaera hakonensis JCM 8857 (1,4-alpha-D-glucosyl)n + alpha,alpha'-trehalose
-
?
3.2.1.141 4'-O-(1,4-alpha-D-glucosyl)n-alpha,alpha'-trehalose + H2O hydrolysis to disaccharides after 24 h Metallosphaera hakonensis JMC8857 (1,4-alpha-D-glucosyl)n + alpha,alpha'-trehalose
-
?

Subunits

EC Number Subunits Comment Organism
3.2.1.141 ? x * 150000, fusion enzyme of maltooligosyltrehalose synthase (MhMTS) and malooligosyltrehalose trehalohydrolase (MhMTH), SDS-PAGE Metallosphaera hakonensis

Synonyms

EC Number Synonyms Comment Organism
3.2.1.141 maltooligosyltrehalose trehalohydrolase
-
Metallosphaera hakonensis
5.4.99.15 MTS
-
Metallosphaera hakonensis
3.2.1.141 MTH
-
Metallosphaera hakonensis
5.4.99.15 maltooligosyltrehalose synthase
-
Metallosphaera hakonensis
5.4.99.15 MTSH fusion enzyme consisting of maltooligosyltrehalose synthase and maltooligosyltrehalose trehalohydrolase Metallosphaera hakonensis
3.2.1.141 MhMTH
-
Metallosphaera hakonensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.141 70
-
-
Metallosphaera hakonensis
5.4.99.15 70
-
MTSH fusion enzyme Metallosphaera hakonensis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.141 40 90 fusion enzyme with 5 mM maltopentaose, 30 min: 100% activity at 70, about 90% at 60, about 80% at 50 and 80, below 40% at 40 and 90°C Metallosphaera hakonensis

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
5.4.99.15 70 80 the MTSH fusion enzyme exhibits a high degree of thermostability, retaining 80% of the activity when pre-incubated at 70°C for 48 h, the stability is gradually abolished by incubating the fusion enzyme at above 80°C for 30 min Metallosphaera hakonensis
3.2.1.141 40 90 80% activity retained after 48 h pre-incubation at 70°C, activities after pre-incubation at set temperatures for 30 min: 90-100% at 40-70°C, less than 80% at 80°C, quickly abolished beyond 80°C Metallosphaera hakonensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.141 5 6
-
Metallosphaera hakonensis
5.4.99.15 5 6 MTSH fusion enzyme Metallosphaera hakonensis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.141 4 9 pH 4: about 70% activity, pH 5-6: about 100% activity, pH 7: about 40% activity, pH 8-9: between 20-30%, pH 4-5: 0.1 M McIlvaine buffer, pH 5-8: 0.1 M sodium phosphate buffer, pH 8-9: 0.1 M sodium bicarbonate buffer, 70°C, 30 min Metallosphaera hakonensis