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Literature summary for 2.4.1.207 extracted from

  • Kallas, A.M.; Piens, K.; Denman, S.E.; Henriksson, H.; Faldt, J.; Johansson, P.; Brumer, H.; Teeri, T.T.
    Enzymatic properties of native and deglycosylated hybrid aspen (Populus tremulaxtremuloides) xyloglucan endotransglycosylase 16A expressed in Pichia pastoris (2005), Biochem. J., 390, 105-113.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
DNA sequence determination and analysis, methanol-induced, functional expression of wild-type and N93S mutant enzymes in Pichia pastoris, the recombinant wild-type enzyme is heterogeneous due to presence of variable N-glycosylation and incomplete cleavage of the alpha-factor secretion signal peptide Populus tremula x Populus tremuloides

Protein Variants

Protein Variants Comment Organism
N93S site-directed mutagenesis, activity and thermal stability of the mutant enzyme are similar to the wild-type enzyme, but the mutant shows reduced solubility Populus tremula x Populus tremuloides

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.062
-
nonasaccharide XLLG-ol pH 5.5, 25°C, recombinant N93S mutant enzyme Populus tremula x Populus tremuloides
0.075
-
nonasaccharide XLLG-ol pH 5.5, 25°C, recombinant wild-type enzyme Populus tremula x Populus tremuloides
1.4
-
xyloglucan pH 5.5, 25°C, recombinant N93S mutant enzyme Populus tremula x Populus tremuloides
2.2
-
xyloglucan pH 5.5, 25°C, recombinant wild-type enzyme Populus tremula x Populus tremuloides

Organism

Organism UniProt Comment Textmining
Populus tremula x Populus tremuloides Q8GZD5
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein enzyme contains a conserved N-glycosylation site situated proximal to the predicted catalytic site, deglycosylation by endoglycosidase H or by site-directed mutagenesis does not influence the enzyme activity Populus tremula x Populus tremuloides

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Pichia pastoris by ultrafiltration, one or two steps of cation exchange chromatography, dialysis, and adsorption chromatography Populus tremula x Populus tremuloides

Reaction

Reaction Comment Organism Reaction ID
2 Xylalpha(1-6)Glcbeta(1-4)(Xylalpha(1-6))Glcbeta(1-4))(Xylalpha(1-6))Glcbeta(1-4)(Xylalpha(1-6))Glcbeta(1-4)Glc = Xylalpha(1-6)Glcbeta(1-4)(Xylalpha(1-6))Glc + Xylalpha(1-6))Glcbeta(1-4)(Xylalpha(1-6))Glcbeta(1-4)Glcbeta(1-4)(Xylalpha(1-6))Glcbeta(1-4)(Xylalpha(1-6))Glcbeta(1-4))(Xylalpha(1-6))Glcbeta(1-4)(Xylalpha(1-6))Glcbeta(1-4)Glc the enzyme contains a catalytic sequence motif conserved in the glycosyl hydrolase family 16 Populus tremula x Populus tremuloides

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.06
-
pH 5.5, 25°C, recombinant N93S mutant enzyme Populus tremula x Populus tremuloides
0.18
-
pH 5.5, 25°C, recombinant wild-type enzyme Populus tremula x Populus tremuloides

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information no hydrolytic activity of the wild-type or N93S mutant enzyme Populus tremula x Populus tremuloides ?
-
?
xyloglucan + nonasaccharide XLLG-ol
-
Populus tremula x Populus tremuloides ?
-
?
xyloglucan nonasaccharide + xyloglucan acceptor: nonasaccharide XLLGol, donor: tamarind xyloglucan Populus tremula x Populus tremuloides ?
-
?

Synonyms

Synonyms Comment Organism
More the enzyme belongs to the glycosyl hydrolase family 16, i.e. GH16 Populus tremula x Populus tremuloides
XET16A
-
Populus tremula x Populus tremuloides
xyloglucan endotransglycosylase 16A
-
Populus tremula x Populus tremuloides

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Populus tremula x Populus tremuloides

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 55 temperature-profile, about 20% of maximal activity at 15°C and 55°C Populus tremula x Populus tremuloides

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
52.3
-
unfolding temperature for the recombinant N93S mutant enzyme, slight protection by 20% glycerol or sucrose, 0.5 M urea, or NaCl at 350 mM, further destabilization by NaCl at 1 M Populus tremula x Populus tremuloides
52.9
-
unfolding temperature for the recombinant wild-type enzyme, slight protection by 20% glycerol or sucrose, 0.5 M urea, or NaCl at 350 mM, further destabilization by NaCl at 1 M Populus tremula x Populus tremuloides

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.4
-
-
Populus tremula x Populus tremuloides

pH Range

pH Minimum pH Maximum Comment Organism
4.5 6.5 pH-profile, rapid loss of activity below pH 4.5 and above pH 6.5 Populus tremula x Populus tremuloides