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Literature summary for 3.2.1.21 extracted from

  • Perugino, G.; Trincone, A.; Giordano, A.; van der Oost, J.; Kaper, T.; Rossi, M.; Moracci, M.
    Activity of hyperthermophilic glycosynthases is significantly enhanced at acidic pH (2003), Biochemistry, 42, 8484-8493.
    View publication on PubMed
Search for more literature for 3.2.1.21

Protein Variants

Protein Variants Comment Organism
N62Y/Q63K/L231F/V303A/A412V mutations observed in all the clones isolated Thermosphaera aggregans
N62Y/Q63K/L231F/V303A/E386G/A412V additional mutation in nucleophile residue E386, mutation completely abolishes activity. The addition of 2 M sodium formate as an external nucleophile leads to the recovery of 4.60% activity with accumulation of oligosaccharides. At pH 3.0 and low concentrations of sodium formate buffer, the hyperthermophilic glycosynthase shows kcat values similar to those of the wild-type and 17fold higher than those observed at the usual reactivation conditions in 2 M sodium formate at pH 6.5 Thermosphaera aggregans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.2
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C Thermosphaera aggregans
0.38
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C, presence of 2 M formate Thermosphaera aggregans
1.01
-
 2-nitrophenyl beta-D-glucopyranoside mutant E387G, pH 6.5, 65°C, presence of 2 M formate Thermosphaera aggregans
2.8
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 6.0, 65°C Thermosphaera aggregans
3.1
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 5.0, 65°C Thermosphaera aggregans
3.3
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 4.0, 65°C Thermosphaera aggregans
6.6
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 3.0, 65°C Thermosphaera aggregans

Organism

Organism UniProt Comment Textmining
Thermosphaera aggregans Q9YGA8
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.39
-
 mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate cellobiose, pH 3.0, 65°C Thermosphaera aggregans
6.3
-
 mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2-nitrophenyl beta-D-galactopyranoside, pH 3.0, 65°C Thermosphaera aggregans
6.4
-
 mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2-nitrophenyl beta-D-xylopyranoside, pH 3.0, 65°C Thermosphaera aggregans
10.4
-
 wild-type, substrate 2-nitrophenyl beta-D-xylopyranoside, pH 3.0, 65°C Thermosphaera aggregans
12
-
 mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 4-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Thermosphaera aggregans
33.8
-
 wild-type, substrate cellobiose, pH 3.0, 65°C Thermosphaera aggregans
61.7
-
 wild-type, substrate 4-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Thermosphaera aggregans
72.2
-
 wild-type, substrate 2-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Thermosphaera aggregans
75
-
 wild-type, substrate 2,4-dinitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Thermosphaera aggregans
129.8
-
 mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2-nitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Thermosphaera aggregans
140.9
-
 wild-type, substrate 2-nitrophenyl beta-D-galactopyranoside, pH 3.0, 65°C Thermosphaera aggregans
207.1
-
 mutant N62Y/Q63K/L231F/V303A/E386G/A412V, substrate 2,4-dinitrophenyl beta-D-glucopyranoside, pH 3.0, 65°C Thermosphaera aggregans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,4-dinitrophenyl beta-D-glucopyranoside + H2O
-
 Thermosphaera aggregans 2,4-dinitrophenol + D-glucopyranose
-
 ?
2-nitrophenyl beta-D-galactopyranoside + H2O
-
 Thermosphaera aggregans 2-nitrophenol + D-galactopyranose
-
 ?
2-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermosphaera aggregans 2-nitrophenol + D-glucopyranose
-
 ?
2-nitrophenyl beta-D-xylopyranoside + H2O
-
 Thermosphaera aggregans 2-nitrophenol + D-xylopyranose
-
 ?
4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Thermosphaera aggregans 4-nitrophenol + D-glucopyranose
-
 ?
cellobiose + H2O
-
 Thermosphaera aggregans 2 D-glucose
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
11.6
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 6.0, 65°C Thermosphaera aggregans
53.9
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 5.0, 65°C Thermosphaera aggregans
72.9
-
 2-nitrophenyl beta-D-glucopyranoside mutant E387G, pH 6.5, 65°C, presence of 2 M formate Thermosphaera aggregans
305
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 4.0, 65°C Thermosphaera aggregans
309
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C Thermosphaera aggregans
439.8
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C, presence of 2 M formate Thermosphaera aggregans
970
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 3.0, 65°C Thermosphaera aggregans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 6.0, 65°C Thermosphaera aggregans
17.2
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 5.0, 65°C Thermosphaera aggregans
72.9
-
 2-nitrophenyl beta-D-glucopyranoside mutant E387G, pH 6.5, 65°C, presence of 2 M formate Thermosphaera aggregans
91
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 4.0, 65°C Thermosphaera aggregans
147.4
-
 2-nitrophenyl beta-D-glucopyranoside mutant N62Y/Q63K/L231F/V303A/E386G/A412V, pH 3.0, 65°C Thermosphaera aggregans
1164
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C, presence of 2 M formate Thermosphaera aggregans
1575
-
 2-nitrophenyl beta-D-glucopyranoside wild-type, pH 6.5, 65°C Thermosphaera aggregans