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Literature summary for 2.4.1.62 extracted from

  • Bernatchez, S.; Gilbert, M.; Blanchard, M.C.; Karwaski, M.F.; Li, J.; Defrees, S.; Wakarchuk, W.W.
    Variants of the beta 1,3-galactosyltransferase CgtB from the bacterium Campylobacter jejuni have distinct acceptor specificities (2007), Glycobiology, 17, 1333-1343.
    View publication on PubMed
Search for more literature for 2.4.1.62

Application

Application Comment Organism
biotechnology the galactosylation of IFNalpha2b[Tn] with an engineered variant of CgtB clearly demonstrates that a bacterial glycosyltransferase is able to glycosylate a human protein in vitro. This is the first time that the direct glycosylation of a human protein by a bacterial glycosyltransferase is reported Campylobacter jejuni

Cloned(Commentary)

Cloned (Comment) Organism
expressed as a MalE fusion protein in Escherichia coli Campylobacter jejuni

Protein Variants

Protein Variants Comment Organism
DELTA20C mutant with deletion of 20 amino acids from carboxy terminus + C-terminal MalE fusion, specific activity (mU/mg): 58.1 (substrate: GalNAcalpha), 127.5 (substrate: GalNAcbeta), 17.5 (substrate: GM2), 374 (substrate: GA2) Campylobacter jejuni
DELTA20N mutant with deletion of 20 amino acids from carboxy terminus + N-terminal MalE fusion, specific activity (mU/mg): 3.3 (substrate: GalNAcalpha), 8.0 (substrate: GalNAcbeta), 2.3 (substrate: GM2), 34.4 (substrate: GA2) Campylobacter jejuni
DELTA30C mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion, specific activity (mU/mg): 11.6 (substrate: GalNAcalpha), 75.7 (substrate: GalNAcbeta), 80.9 (substrate: GM2) Campylobacter jejuni
DELTA30C mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion, specific activity (mU/mg): 531.5 (substrate: GalNAcalpha), 1299.5 (substrate: GalNAcbeta), 2420 (substrate: GM2), 271.5 (substrate: IFNalpha2b). kcat (1/sec): 0.5 (GalNAcalpha-FCHASE), 1.07 (GalNAcbeta-FCHASE), 0.05 (UDP-Gal), Km (mM): 1.68 (GalNAcalpha-FCHASE), 3.18 (GalNAcbeta-FCHASE), 1.11 (UDP-Gal) Campylobacter jejuni
DELTA30N mutant with deletion of 30 amino acids from carboxy terminus + N-terminal MalE fusion, specific activity (mU/mg): 0.9 (substrate: GalNAcalpha), 5.3 (substrate: GalNAcbeta), 14 (substrate: GM2) Campylobacter jejuni
DELTA30N mutant with deletion of 30 amino acids from carboxy terminus + N-terminal MalE fusion, specific activity (mU/mg): 49.5 (substrate: GalNAcalpha), 36 (substrate: GalNAcbeta), 96 (substrate: GM2), 14 (substrate: IFNalpha2b) Campylobacter jejuni

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.11
-
 UDP-Gal using fusion construct CgtBOH4384DELTA30-MalE (mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion) Campylobacter jejuni
1.68
-
 GalNAcalpha-FCHASE using fusion construct CgtBOH4384DELTA30-MalE (mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion) Campylobacter jejuni
3.18
-
 GalNAcbeta-FCHASE using fusion construct CgtBOH4384DELTA30-MalE (mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion) Campylobacter jejuni

Organism

Organism UniProt Comment Textmining
Campylobacter jejuni Q0P9B5
-
-
Campylobacter jejuni Q93MQ1
-
-
Campylobacter jejuni Q9K2V8
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 relative activity: GalNAcalpha: 1.4%, GalNAcbeta: 1.9%, GM2: 38%, lyso-GM2: 0.3%, GA2: 100% Campylobacter jejuni
additional information
-
 relative activity: GalNAcalpha: 3.0%, GalNAcbeta: 50.5%, GM2: 100%, lyso-GM2: 606%, GA2: 8.8% Campylobacter jejuni
additional information
-
 relative activity: GalNAcalpha: 6.4%, GalNAcbeta: 47.5%, GM2: 100%, lyso-GM2: 444%, GA2: 29% Campylobacter jejuni
additional information
-
 specific activity of N-term MalE-CgtBOH4384 fusion construct (mU/mg): 158 (substrate: GalNAcalpha), 192 (substrate: GalNAcbeta), 210 (substrate: GM2), 83.5 (substrate: IFNalpha2b) Campylobacter jejuni
additional information
-
 specific activity of N-term MalE-CgtBOH4384 fusion construct (mU/mg): 9.7 (substrate: GalNAcalpha), 4.8 (substrate: GalNAcbeta), 4.5 (substrate: GM2), 86.9 (substrate: GA2) Campylobacter jejuni
additional information
-
 specific activity of N-term MalE–CgtBOH4384 fusion construct (mU/mg): 13.6 (substrate: GalNAcalpha), 40.4 (substrate: GalNAcbeta), 27 (substrate: GM2) Campylobacter jejuni

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-Gal + GA2-FCHASE FCHASE: 6-(fluorescein-5-carboxamido)hexanoic acid succinimidyl ester Campylobacter jejuni ?
-
 ?
UDP-Gal + GalNAcalpha-FCHASE FCHASE: 6-(fluorescein-5-carboxamido)hexanoic acid succinimidyl ester Campylobacter jejuni ?
-
 ?
UDP-Gal + GalNAcbeta-FCHASE FCHASE: 6-(fluorescein-5-carboxamido)hexanoic acid succinimidyl ester Campylobacter jejuni ?
-
 ?
UDP-Gal + GM2-FCHASE FCHASE: 6-(fluorescein-5-carboxamido)hexanoic acid succinimidyl ester Campylobacter jejuni ?
-
 ?
UDP-Gal + IFNalpha2b-[Tn]-FCHASE the galactosylation of human therapeutic protein IFNalpha2b[Tn] with an engineered variant of CgtB clearly demonstrates that a bacterial glycosyltransferase is able to glycosylate a human protein in vitro. FCHASE: 6-(fluorescein-5-carboxamido)hexanoic acid succinimidyl ester Campylobacter jejuni ?
-
 ?
UDP-Gal + lyso-GM2-FCHASE FCHASE: 6-(fluorescein-5-carboxamido)hexanoic acid succinimidyl ester Campylobacter jejuni ?
-
 ?

Synonyms

Synonyms Comment Organism
beta 1,3-galactosyltransferase
-
 Campylobacter jejuni
CgtB 11168
-
 Campylobacter jejuni
cgtBOH HS:10
-
 Campylobacter jejuni
cgtBOH4384
-
 Campylobacter jejuni

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Campylobacter jejuni

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.05
-
 UDP-Gal using fusion construct CgtBOH4384DELTA30-MalE (mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion) Campylobacter jejuni
0.5
-
 GalNAcalpha-FCHASE using fusion construct CgtBOH4384DELTA30-MalE (mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion) Campylobacter jejuni
1.07
-
 GalNAcbeta-FCHASE using fusion construct CgtBOH4384DELTA30-MalE (mutant with deletion of 30 amino acids from carboxy terminus + C-terminal MalE fusion) Campylobacter jejuni

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 assay at Campylobacter jejuni