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Literature summary for 3.2.1.8 extracted from

  • Ribeiro, L.F.; Furtado, G.P.; Lourenzoni, M.R.; Costa-Filho, A.J.; Santos, C.R.; Nogueira, S.C.; Betini, J.A., Polizeli, Mde L.; Murakami, M.T.; Ward, R.J.
    Engineering bifunctional laccase-xylanase chimeras for improved catalytic performance (2011), J. Biol. Chem., 286, 43026-4338.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.2.1.8

Cloned(Commentary)

Cloned (Comment) Organism
the chimeric enzymes, CorA-XynA and CorA-XynAG3, the parental enzyme XynA, and the thermostable variant XynAG3 are produced in Escherichia coli Bacillus subtilis

Protein Variants

Protein Variants Comment Organism
additional information the fusion of the 1642-bp laccase (CorA) with either the 555-bp xylanase (XynA) or the thermostable variant (XynAG3) are performed by insertion of the xylanase into a surface loop of the laccase. The resulting chimeric constructs of 2197 bp contain a central region composed of the XynA or XynAG3 sequence flanked by the regions of the CorA encoding the N-terminal residues 1–216 (forming the 5' region of the chimera) and the C-terminal region comprising residues 217–513 of CorA. As a consequence, the final construct results in two linkage points between the laccase and xylanase domains Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 1,4-beta-D-xylan Km for chimeric enzyme CorA-XynA at pH 6.5, 65°C: 6.3 mg/ml, KM for chimeric enzyme CorA-XynAG3 at pH 7.5, 75°C: 5.3 mg/ml, Km for xylanase XynA at pH 6.5, 55°C: 5.5 mg/ml, Km for thermostabel xylanase variant XynAG3 at pH 7.5, 70°C: 5.1 mg/ml Bacillus subtilis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
23000
-
 parental enzyme XynA, SDS-PAGE Bacillus subtilis
85500
-
 chimeric enzymes CorA-XynA and CorA-XynAG3, SDS-PAGE Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
the chimeric enzymes, CorA-XynA and CorA-XynAG3, the parental enzyme XynA, and the thermostable variant XynAG3 are produced in Escherichia coli Bacillus subtilis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1,4-beta-D-xylan + H2O
-
 Bacillus subtilis ?
-
 ?

Synonyms

Synonyms Comment Organism
endo-1,4-beta-xylanase
-
 Bacillus subtilis
XynA
-
 Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
 xylanase XynA Bacillus subtilis
65
-
 chimeric enzyme CorAXynA Bacillus subtilis
70
-
 thermostable variant XynAG3 and chimeric enzyme CorAXynAG3 Bacillus subtilis

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
 the thermostable variant XynAG3 loses 90% of its activity after 10 min, and after 30 min no activity is observed. The chimeric CotA-XynAG3 retains 60% activity after 10 min and about 20% activity after 120 min. The half-lives of the chimeric enzymes XynAG3 and CotA-XynAG3 are 2.2 and 21 min Bacillus subtilis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
61
-
 1,4-beta-D-xylan pH 7.5, 70°C, thermostable xylanase variant XynAG3, calculated as D-xylose liberated from 1,4-beta-D-xylan per second Bacillus subtilis
65
-
 1,4-beta-D-xylan pH 7.5, 70°C, chimeric enzyme CorA-XynAG3, calculated as D-xylose liberated from 1,4-beta-D-xylan per second Bacillus subtilis
75
-
 1,4-beta-D-xylan pH 6.5, 65°C, chimeric enzyme CorA-XynA, calculated as D-xylose liberated from 1,4-beta-D-xylan per second Bacillus subtilis
82
-
 1,4-beta-D-xylan pH 6.5, 55°C, xylanase XynA, calculated as D-xylose liberated from 1,4-beta-D-xylan per second Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
 xylanase XynA and chimeric enzyme CorA-XynA Bacillus subtilis
7.5
-
 thermostabel xylanase variant XynAG3 and chimeric enzyme CorA-XynAG3 Bacillus subtilis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
additional information
-
 1,4-beta-D-xylan kcat/Km for chimeric enzyme CorA-XynA at pH 6.5, 65°C: 12 ml/mg*ml, kcat/KM for chimeric enzyme CorA-XynAG3 at pH 7.5, 75°C: 12 ml/mg*ml, kcat/Km for xylanase XynA at pH 6.5, 55°C: 14.9 ml/mg*ml, kcat/Km for thermostabel xylanase variant XynAG3 at pH 7.5, 70°C: 11.9 ml/mg*ml Bacillus subtilis