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Literature summary extracted from

  • Steiner, K.; Novotny, R.; Patel, K.; Vinogradov, E.; Whitfield, C.; Valvano, M.A.; Messner, P.; Schaeffer, C.
    Functional characterization of the initiation enzyme of S-layer glycoprotein glycan biosynthesis in Geobacillus stearothermophilus NRS 2004/3a (2007), J. Bacteriol., 189, 2590-2598.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.8.6 expressed in Escherichia coli TOP10 and BL21(DE3) Star cells, and in Salmonella enterica serovar Typhimurium Geobacillus stearothermophilus
2.7.8.6 from genomic DNA, (i) in pET28a for inducible expression of entire coding sequence or C-terminal part with N-terminal hexa-His-tag and subcellular localisation in Escherichia coli BL21(DE3) Star, (ii) in pBAD-DEST49: for inducible expression with N-terminal horseradish peroxide-thioredoxin-tag and C-terminal hexa-His-tag in Escherichia coli TOP10, (iii) in pBluescript II SK(+): for complementation studies with full-length WsaP, C-terminal part (aa168-417), N-terminal part (aa1-314) or a mutant lacking the transmembrane domains (aa311-471) in Escherichia coli strain CWG466 and Salmonella enterica strain MSS2 Geobacillus stearothermophilus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.8.6 membrane
-
 Geobacillus stearothermophilus 16020
-
2.7.8.6 membrane membrane fraction, differential centrifugation, Western blot Geobacillus stearothermophilus 16020
-
2.7.8.6 additional information no inclusion bodies Geobacillus stearothermophilus
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.8.6 36500
-
 theoretical and apparent for hexa-His-tagged C-terminal part (aa168-417), SDS-PAGE Geobacillus stearothermophilus
2.7.8.6 41800
-
 hexa-His-tagged full-length enzyme, SDS-PAGE/Western blot Geobacillus stearothermophilus
2.7.8.6 56800
-
 SDS-PAGE Geobacillus stearothermophilus
2.7.8.6 56800
-
 theoretical for hexa-His-tagged full-length enzyme Geobacillus stearothermophilus
2.7.8.6 72870
-
 theoretical for full-length with N-terminal horseradish peroxide-thioredoxin-tag and C-terminal hexa-His-tag, apparent weight slightly less in SDS-PAGE Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.8.6 additional information Geobacillus stearothermophilus initiation step in S-layer glycoprotein glycan biosynthesis ?
-
 ?
2.7.8.6 additional information Geobacillus stearothermophilus NRS 2004/3a initiation step in S-layer glycoprotein glycan biosynthesis ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.8.6 Geobacillus stearothermophilus
-
-
-
2.7.8.6 Geobacillus stearothermophilus Q7BG44 NRS 2004/3a
-
2.7.8.6 Geobacillus stearothermophilus NRS 2004/3a
-
-
-
2.7.8.6 Geobacillus stearothermophilus NRS 2004/3a Q7BG44 NRS 2004/3a
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.8.6 from Escherichia coli BL21(DE3) Star and TOP10 membrane preparations, nickel affinity chromatography (batch method), in presence of 0.1 mM lauryldimethylamine N-oxide Geobacillus stearothermophilus
2.7.8.6 His-Select nickel affinity gel column chromatography Geobacillus stearothermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.8.6 additional information initiation step in S-layer glycoprotein glycan biosynthesis Geobacillus stearothermophilus ?
-
 ?
2.7.8.6 additional information K30 antigen biosynthesis by full-length enzyme and N-terminal truncated version, membrane fraction Geobacillus stearothermophilus ?
-
 ?
2.7.8.6 additional information initiation step in S-layer glycoprotein glycan biosynthesis Geobacillus stearothermophilus NRS 2004/3a ?
-
 ?
2.7.8.6 additional information K30 antigen biosynthesis by full-length enzyme and N-terminal truncated version, membrane fraction Geobacillus stearothermophilus NRS 2004/3a ?
-
 ?
2.7.8.6 UDP-D-galactose + phosphoryl-polyprenol glycosyltransferase activity within C-terminus of WsaP, in vitro assay: 1 h, 37°C, pH 8.5, 1 mM EDTA, 10 mM MgCl2, membrane fraction Geobacillus stearothermophilus galactosyl-diphospho-polyprenol + CMP analyses by thin-layer chromatography ?
2.7.8.6 UDP-D-galactose + phosphoryl-polyprenol glycosyltransferase activity within C-terminus of WsaP, in vitro assay: 1 h, 37°C, pH 8.5, 1 mM EDTA, 10 mM MgCl2, membrane fraction Geobacillus stearothermophilus NRS 2004/3a galactosyl-diphospho-polyprenol + CMP analyses by thin-layer chromatography ?
2.7.8.6 UDP-galactose + phosphoryl-polyprenol
-
 Geobacillus stearothermophilus UMP + alpha-D-galactosyl-diphosphopolyprenol
-
 ?
2.7.8.6 UDP-galactose + phosphoryl-polyprenol
-
 Geobacillus stearothermophilus NRS 2004/3a UMP + alpha-D-galactosyl-diphosphopolyprenol
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.8.6 UDP-Gal: phosphoryl-polyprenol Gal-1-phosphate transferase
-
 Geobacillus stearothermophilus
2.7.8.6 UDP-Gal:phosphoryl-polyprenol Gal-1-phosphate transferase
-
 Geobacillus stearothermophilus
2.7.8.6 WsaP
-
 Geobacillus stearothermophilus
2.7.8.6 WsaP the enzyme transfers in vitro a galactose-1-phosphate from UDP-galactose to endogenous phosphoryl-polyprenol Geobacillus stearothermophilus