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 | 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Populus tremula x Populus tremuloides | Q8GZD5 | - |
- |
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 (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 | 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 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 | 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 | 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 Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
- |
Populus tremula x Populus tremuloides |
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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.4 | - |
- |
Populus tremula x Populus tremuloides |
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 |