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Literature summary for 2.4.1.20 extracted from

  • Wu, Y.; Mao, G.; Fan, H.; Song, A.; Zhang, Y.P.; Chen, H.
    Biochemical properties of GH94 cellodextrin phosphorylase THA_1941 from a thermophilic eubacterium Thermosipho africanus TCF52B with cellobiose phosphorylase activity (2017), Sci. Rep., 7, 4849 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Thermosipho africanus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.06
-
cellopentaose at pH 7.5 and 75°C Thermosipho africanus
0.062
-
cellotetraose at pH 7.5 and 75°C Thermosipho africanus
0.094
-
cellotriose at pH 7.5 and 75°C Thermosipho africanus
0.12
-
cellobiose at pH 7.5 and 75°C Thermosipho africanus
1.57
-
alpha-D-glucose 1-phosphate at pH 7.5 and 75°C Thermosipho africanus
14.3
-
D-glucose at pH 7.5 and 75°C Thermosipho africanus
25.7
-
D-xylose at pH 7.5 and 75°C Thermosipho africanus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ 1 mM used in assay conditions Thermosipho africanus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
alpha-D-glucose 1-phosphate + D-glucose Thermosipho africanus
-
cellobiose + phosphate
-
r
cellobiose + phosphate Thermosipho africanus
-
alpha-D-glucose 1-phosphate + D-glucose
-
r

Organism

Organism UniProt Comment Textmining
Thermosipho africanus B7IED6
-
-
Thermosipho africanus TCF52B B7IED6
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-chelating column chromatography and HiTrap Q column chromatography Thermosipho africanus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.16
-
with D-maltose as substrate, at pH 7.5 and 75°C Thermosipho africanus
0.22
-
with D-arabinose as substrate, at pH 7.5 and 75°C Thermosipho africanus
0.22
-
with D-fructose as substrate, at pH 7.5 and 75°C Thermosipho africanus
0.22
-
with N-acetyl-D-glucosamine as substrate, at pH 7.5 and 75°C Thermosipho africanus
0.27
-
with D-galactose as substrate, at pH 7.5 and 75°C Thermosipho africanus
0.35
-
with D-xylose as substrate, at pH 7.5 and 75°C Thermosipho africanus
0.38
-
with D-mannose as substrate, at pH 7.5 and 75°C Thermosipho africanus
0.85
-
with 2-deoxy-D-glucose as substrate, at pH 7.5 and 75°C Thermosipho africanus
1.08
-
with D-glucosamine as substrate, at pH 7.5 and 75°C Thermosipho africanus
1.86
-
with D-glucose as substrate, at pH 7.5 and 75°C Thermosipho africanus
49.98
-
with gentiobiose as substrate, at pH 7.5 and 75°C Thermosipho africanus
58.61
-
with cellobiose as substrate, at pH 7.5 and 75°C Thermosipho africanus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-D-glucose 1-phosphate + 2-deoxy-D-glucose
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + D-arabinose
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + D-fructose
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + D-galactose
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + D-glucosamine
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + D-glucose
-
Thermosipho africanus cellobiose + phosphate
-
r
alpha-D-glucose 1-phosphate + D-maltose
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + D-mannose
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + D-xylose
-
Thermosipho africanus phosphate + ?
-
r
alpha-D-glucose 1-phosphate + N-acetyl-D-glucosamine
-
Thermosipho africanus phosphate + ?
-
r
cellobiose + phosphate
-
Thermosipho africanus alpha-D-glucose 1-phosphate + D-glucose
-
r
cellobiose + phosphate 38.6% activity compared to cellotetraose Thermosipho africanus alpha-D-glucose 1-phosphate + D-glucose
-
r
cellopentaose + phosphate 63.3% activity compared to cellotetraose Thermosipho africanus ?
-
r
cellotetraose + phosphate 100% activity Thermosipho africanus ?
-
r
cellotriose + phosphate 89.7% activity compared to cellotetraose Thermosipho africanus ?
-
r
gentiobiose + phosphate
-
Thermosipho africanus ?
-
r
additional information beyond its cellobiose phosphorylase activity, this enzyme can use cellobiose and long-chain cellodextrins with a degree of polymerization of greater than 2 as a glucose acceptor, releasing phosphate from alpha-D-glucose 1-phosphate Thermosipho africanus ?
-
-

Subunits

Subunits Comment Organism
? x * 120000, SDS-PAGE Thermosipho africanus

Synonyms

Synonyms Comment Organism
Cbp
-
Thermosipho africanus
THA_1941
-
Thermosipho africanus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
75
-
-
Thermosipho africanus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
50 90 the enzyme shows more than 50% activity between 50 and 90°C Thermosipho africanus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
75
-
the enzyme retains 80% of its initial activity after incubating at 75°C and pH 7.5 for 30 min Thermosipho africanus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.23
-
cellobiose at pH 7.5 and 75°C Thermosipho africanus
0.28
-
cellotriose at pH 7.5 and 75°C Thermosipho africanus
0.34
-
cellopentaose at pH 7.5 and 75°C Thermosipho africanus
0.49
-
cellotetraose at pH 7.5 and 75°C Thermosipho africanus
1.37
-
D-xylose at pH 7.5 and 75°C Thermosipho africanus
7.61
-
alpha-D-glucose 1-phosphate at pH 7.5 and 75°C Thermosipho africanus
30.3
-
D-glucose at pH 7.5 and 75°C Thermosipho africanus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Thermosipho africanus

pH Range

pH Minimum pH Maximum Comment Organism
4 9.5 the enzyme shows more than 50% activity between pH 4.0 and 9.5 Thermosipho africanus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.05
-
D-xylose at pH 7.5 and 75°C Thermosipho africanus
1.92
-
cellobiose at pH 7.5 and 75°C Thermosipho africanus
2.12
-
D-glucose at pH 7.5 and 75°C Thermosipho africanus
2.98
-
cellotriose at pH 7.5 and 75°C Thermosipho africanus
4.85
-
alpha-D-glucose 1-phosphate at pH 7.5 and 75°C Thermosipho africanus
5.67
-
cellopentaose at pH 7.5 and 75°C Thermosipho africanus
7.91
-
cellotetraose at pH 7.5 and 75°C Thermosipho africanus