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Literature summary for 1.1.5.2 extracted from

  • Hofer, M.; Boensch, K.; Greiner-Stoeffele, T.; Ballschmiter, M.
    Characterization and engineering of a novel pyrroloquinoline quinone dependent glucose dehydrogenase from Sorangium cellulosum So ce56 (2011), Mol. Biotechnol., 47, 253-261.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli NovaBlue (DE3) cells Sorangium cellulosum

Protein Variants

Protein Variants Comment Organism
Q126E the mutant shows 25.6% relative activity on maltose Sorangium cellulosum
Q126R the mutant shows 11.4% relative activity on maltose Sorangium cellulosum
Q126S the mutant shows 12.4% relative activity on maltose Sorangium cellulosum
Q219E/F220E the mutant shows 5.2% relative activity on maltose Sorangium cellulosum
Q219K/F220C the mutant shows 20% relative activity on maltose Sorangium cellulosum
Q219K/F220K the mutant shows 29.3% relative activity on maltose Sorangium cellulosum
Q219N/F220K the mutant shows 11.7% relative activity on maltose Sorangium cellulosum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4
-
D-glucose wild type enzyme, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
7.2
-
D-glucose mutant enzyme Q219K/F220C, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
7.6
-
D-glucose mutant enzyme Q219K/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
10.6
-
D-glucose mutant enzyme Q219N/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
11
-
D-glucose mutant enzyme Q126E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
11.3
-
D-glucose mutant enzyme Q219E/F220E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
22.6
-
D-glucose mutant enzyme Q126S, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
41.4
-
D-glucose mutant enzyme Q126R, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ Ca2+ is needed for the correct folding and for the functionalization of pyrroloquinoline quinone as cofactor Sorangium cellulosum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
56000
-
calculated from amino acid sequence Sorangium cellulosum
63000
-
recombinant enzyme, SDS-PAGE Sorangium cellulosum

Organism

Organism UniProt Comment Textmining
Sorangium cellulosum
-
-
-
Sorangium cellulosum So ce56
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography, gel filtration Sorangium cellulosum

Renatured (Commentary)

Renatured (Comment) Organism
protein refolding is started on the column during elution in the presence of Ca2+. Refolding is finished by dialysis against 50 mM sodium potassium phosphate buffer pH 7.5 containing 0.003 mM CaCl2 and 0.002 mM pyrroloquinoline quinone. Refolding without Ca2+ and pyrroloquinoline quinone leads to an inactive enzyme Sorangium cellulosum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-deoxy-D-glucose + 2,6-dichlorophenolindophenol 3% activity compared to D-glucose Sorangium cellulosum 2-deoxy-D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
2-deoxy-D-glucose + 2,6-dichlorophenolindophenol 3% activity compared to D-glucose Sorangium cellulosum So ce56 2-deoxy-D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-allose + 2,6-dichlorophenolindophenol 85% activity compared to D-glucose Sorangium cellulosum D-allono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-allose + 2,6-dichlorophenolindophenol 85% activity compared to D-glucose Sorangium cellulosum So ce56 D-allono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-galactose + 2,6-dichlorophenolindophenol 71% activity compared to D-glucose Sorangium cellulosum D-galactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-galactose + 2,6-dichlorophenolindophenol 71% activity compared to D-glucose Sorangium cellulosum So ce56 D-galactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-glucose + 2,6-dichlorophenolindophenol 100% activity Sorangium cellulosum D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-glucose + 2,6-dichlorophenolindophenol 100% activity Sorangium cellulosum So ce56 D-glucono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-lactose + 2,6-dichlorophenolindophenol 91% activity compared to D-glucose Sorangium cellulosum D-lactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-lactose + 2,6-dichlorophenolindophenol 91% activity compared to D-glucose Sorangium cellulosum So ce56 D-lactono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-maltose + 2,6-dichlorophenolindophenol 71% activity compared to D-glucose Sorangium cellulosum D-maltono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-mannose + 2,6-dichlorophenolindophenol 36% activity compared to D-glucose Sorangium cellulosum D-mannono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
D-xylose + 2,6-dichlorophenolindophenol 86% activity compared to D-glucose Sorangium cellulosum D-xylono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?
L-arabinose + 2,6-dichlorophenolindophenol 97% activity compared to D-glucose Sorangium cellulosum L-arabinono-1,5-lactone + reduced 2,6-dichlorophenolindophenol
-
?

Synonyms

Synonyms Comment Organism
PQQ GDH
-
Sorangium cellulosum
pyrroloquinoline quinone dependent glucose dehydrogenase
-
Sorangium cellulosum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
-
Sorangium cellulosum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
25 40 at 25°C the enzyme activity is reduced to 50%, whereas at 40°C the activity is only reduced to 90% compared to the activity at 37°C Sorangium cellulosum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 65 100% activity at 25-30°C, about 90% activity at 35°C, about 80% activity at 40-45°C, about 750% activity at 50°C, about 60% activity at 55°C, after incubation at 60°C for 1 h the enzyme still has 27% of its initial activity while loosing nearly all activity after treatment at 65°C Sorangium cellulosum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
121.5
-
D-glucose mutant enzyme Q219K/F220C, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
197.1
-
D-glucose mutant enzyme Q126E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
227.2
-
D-glucose mutant enzyme Q219N/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
305.9
-
D-glucose mutant enzyme Q219E/F220E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
341.6
-
D-glucose mutant enzyme Q126R, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
445.1
-
D-glucose mutant enzyme Q126S, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
1114
-
D-glucose mutant enzyme Q219K/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
1779
-
D-glucose wild type enzyme, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
the highest activity is observed with 42 mM sodium potassium phosphate buffer at pH 7.5 Sorangium cellulosum

pH Range

pH Minimum pH Maximum Comment Organism
5.5 9
-
Sorangium cellulosum

Cofactor

Cofactor Comment Organism Structure
pyrroloquinoline quinone required for activity Sorangium cellulosum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
8.2
-
D-glucose mutant enzyme Q126R, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
16.8
-
D-glucose mutant enzyme Q219K/F220C, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
18
-
D-glucose mutant enzyme Q126E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
19.7
-
D-glucose mutant enzyme Q126S, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
21.7
-
D-glucose mutant enzyme Q219N/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
27
-
D-glucose mutant enzyme Q219E/F220E, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
146.6
-
D-glucose mutant enzyme Q219K/F220K, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum
744
-
D-glucose wild type enzyme, in 42 mM sodium phosphate buffer pH 7.5 at 37°C Sorangium cellulosum