Cloned (Comment) | Organism |
---|---|
expression of laccase 3 variants in Saccharomyces cerevisiae | Trametes sp. |
Protein Variants | Comment | Organism |
---|---|---|
additional information | a variant deleted of the last 13 residues (C(DELTA)) and its 6His tagged counterpart (C(DELTA)6H) are active enzymes. The production of C(DELTA)6H results in the synthesis of a unusually high proportion of highly glycosylated forms of the enzyme therefore allowing the additional purification of a hyper-glycosylated form of C(DELTA)6H noted C(DELTA)6Hh. Properties of (C(DELTA)), C(DELTA)6H and C(DELTA)6Hh are compared. Catalytic efficiency of C(DELTA) (kcat/KM) decreases 4fold (with syringaldazine as substrate) and 10fold (with 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) as substrate) respectively. The catalytic parameters kcat and KM of C(DELTA)6H and C(DELTA)6Hh are strictly comparable revealing that over-glycosylation does not affect the enzyme catalytic efficiency. To the contrary, in vitro deglycosylation of laccase drastically reduces its activity | Trametes sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0072 | - |
syringaldazine | 30°C, pH 6.0, C-6His tagged laccase 3 | Trametes sp. | |
0.0081 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) | Trametes sp. | |
0.0093 | - |
syringaldazine | 30°C, pH 6.0, deglycosylated enzyme | Trametes sp. | |
0.0116 | - |
syringaldazine | 30°C, pH 6.0, wild-type enzyme | Trametes sp. | |
0.0126 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) and hyper-glycosylated | Trametes sp. | |
0.0237 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) | Trametes sp. | |
0.722 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, C-6His tagged laccase 3 | Trametes sp. | |
0.835 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, deglycosylated enzyme | Trametes sp. | |
0.964 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) | Trametes sp. | |
1.237 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) | Trametes sp. | |
1.268 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)), 6His tagged (C(DELTA)6H) and hyper-glycosylated | Trametes sp. | |
2.042 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, wild-type enzyme | Trametes sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trametes sp. | Q6TH77 | - |
- |
Trametes sp. C30 | Q6TH77 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | in vitro deglycosylation of laccase drastically reduces its activity | Trametes sp. |
Purification (Comment) | Organism |
---|---|
- |
Trametes sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) + O2 | - |
Trametes sp. | ? | - |
? | |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) + O2 | - |
Trametes sp. C30 | ? | - |
? | |
syringaldazine + O2 | - |
Trametes sp. | ? | - |
? | |
syringaldazine + O2 | - |
Trametes sp. C30 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 80000, SDS-PAGE | Trametes sp. |
Synonyms | Comment | Organism |
---|---|---|
LAC3 | - |
Trametes sp. |
laccase 3 | - |
Trametes sp. |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
thermal stability of the enzyme is negatively influenced by an alteration of the amino acid sequence at the C terminus. Glycosylation enhances the thermal stability of the enzyme | Trametes sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
67 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) | Trametes sp. | |
71 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) | Trametes sp. | |
113 | - |
syringaldazine | 30°C, pH 6.0, C-6His tagged laccase 3 | Trametes sp. | |
170 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, C-6His tagged laccase 3 | Trametes sp. | |
188 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) | Trametes sp. | |
246 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) and hyper-glycosylated | Trametes sp. | |
290 | - |
syringaldazine | 30°C, pH 6.0, deglycosylated enzyme | Trametes sp. | |
299 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) | Trametes sp. | |
328 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)), 6His tagged (C(DELTA)6H) and hyper-glycosylated | Trametes sp. | |
386 | - |
syringaldazine | 30°C, pH 6.0, wild-type enzyme | Trametes sp. | |
445 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, deglycosylated enzyme | Trametes sp. | |
549 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, wild-type enzyme | Trametes sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
substrate: syringaldazine | Trametes sp. |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
2.2 | - |
unstable at | Trametes sp. |
8 | - |
24 h, residual activity of laccase 3 is 89%, residual activity of deglycosylated laccase 3 is 77% | Trametes sp. |
9 | - |
unstable at | Trametes sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
70 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) | Trametes sp. | |
240 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) | Trametes sp. | |
240 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, deglycosylated enzyme | Trametes sp. | |
260 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)), 6His tagged (C(DELTA)6H) and hyper-glycosylated | Trametes sp. | |
270 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, wild-type enzyme | Trametes sp. | |
530 | - |
2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) | 30°C, pH 6.0, C-6His tagged laccase 3 | Trametes sp. | |
3000 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) | Trametes sp. | |
16000 | - |
syringaldazine | 30°C, pH 6.0, deglycosylated enzyme | Trametes sp. | |
20000 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) and hyper-glycosylated | Trametes sp. | |
23000 | - |
syringaldazine | 30°C, pH 6.0, a variant of laccase 3 deleted of the last 13 residues (C(DELTA)) and 6His tagged (C(DELTA)6H) | Trametes sp. | |
31000 | - |
syringaldazine | 30°C, pH 6.0, C-6His tagged laccase 3 | Trametes sp. | |
33000 | - |
syringaldazine | 30°C, pH 6.0, wild-type enzyme | Trametes sp. |