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Literature summary for 1.10.3.2 extracted from

  • Liu, Y.; Cusano, A.M.; Wallace, E.C.; Mekmouche, Y.; Ullah, S.; Robert, V.; Tron, T.
    Characterization of C-terminally engineered laccases (2014), Int. J. Biol. Macromol., 69, 435-441 .
    View publication on PubMed
Search for more literature for 1.10.3.2

Cloned(Commentary)

Cloned (Comment) Organism
expression of laccase 3 variants in Saccharomyces cerevisiae Trametes sp.

Protein Variants

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 [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

Organism UniProt Comment Textmining
Trametes sp. Q6TH77
-
-
Trametes sp. C30 Q6TH77
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein in vitro deglycosylation of laccase drastically reduces its activity Trametes sp.

Purification (Commentary)

Purification (Comment) Organism
-
 Trametes sp.

Substrates and Products (Substrate)

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

Subunits Comment Organism
? x * 80000, SDS-PAGE Trametes sp.

Synonyms

Synonyms Comment Organism
LAC3
-
 Trametes sp.
laccase 3
-
 Trametes sp.

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5
-
 substrate: syringaldazine Trametes sp.

pH Stability

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 [mM/s]

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.