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Literature summary for 3.2.1.14 extracted from

  • Boer, H.; Simolin, H.; Cottaz, S.; Soederlund, H.; Koivula, A.
    Heterologous expression and site-directed mutagenesis studies of two Trichoderma harzianum chitinases, Chit33 and Chit42, in Escherichia coli (2007), Protein Expr. Purif., 51, 216-226.
    View publication on PubMed
Search for more literature for 3.2.1.14

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli. Cytoplasmic overexpression results in inclusion body formation. Active enzyme can be recovered after refolding. Periplasmic proves to be better suited for mutagenesis purposes Trichoderma harzianum

Protein Variants

Protein Variants Comment Organism
D170N 0.6% of wild-type activity Trichoderma harzianum
E172Q 0.3% of wild-type activity Trichoderma harzianum

Inhibitors

Inhibitors Comment Organism Structure
allosamidin
-
 Trichoderma harzianum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.9
-
 4-nitrophenyl 2-(acetylamino)-4-O-[2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl]-2-deoxy-beta-D-glucopyranoside pH 6.5, wild-type enzyme from Trichoderma harzianum Trichoderma harzianum
1.1
-
 4-nitrophenyl 2-(acetylamino)-4-O-[2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl]-2-deoxy-beta-D-glucopyranoside pH 6.5, wilde-type recombinant enzyme expressed in Escherichia coli Trichoderma harzianum

Organism

Organism UniProt Comment Textmining
Trichoderma harzianum P48827
-
-
Trichoderma harzianum Q12713
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
additional information Chit33 does not contain any putative N-glycosylation site Trichoderma harzianum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-N,N'-diacetylchitobioside + H2O essential role of the two carboxylic acid residues E172 and D170 in the catalytic mechanism of Chit42 Trichoderma harzianum ?
-
 ?
additional information essential role of E172, D170 and E145 in the catalytic mechanism of Chit33 Trichoderma harzianum ?
-
 ?

Synonyms

Synonyms Comment Organism
Chit33
-
 Trichoderma harzianum
CHIT42
-
 Trichoderma harzianum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
38
-
 enzyme form Trichoderma harzianum Trichoderma harzianum
47
-
 enzyme recombinantly expressed in Escherichia coli and refolded Trichoderma harzianum
50
-
 enzyme recombinantly expressed in Escherichia coli and refolded Trichoderma harzianum
55
-
 enzyme form Trichoderma harzianum Trichoderma harzianum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
23 50 23°C: about 50% of maximal activity, 50°C: about 30% of maximal activity, enzyme form Trichoderma harzianum Trichoderma harzianum
23 50 23°C: about 60% of maximal activity, 50°C: about 30% of maximal activity, enzyme recombinantly expressed in Escherichia coli and refolded Trichoderma harzianum
40 55 40°C: about 80% of maximal activity, 55°C: about 90% of maximal activity, 65°C: no activity, enzyme form Trichoderma harzianum Trichoderma harzianum
40 60 40°C: about 70% of maximal activity, 60°C: about 80% of maximal activity, 70°C, no activity, enzyme recombinantly expressed in Escherichia coli and refolded Trichoderma harzianum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.267
-
 4-nitrophenyl 2-(acetylamino)-4-O-[2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl]-2-deoxy-beta-D-glucopyranoside pH 6.5, wild-type enzyme from Trichoderma harzianum Trichoderma harzianum
0.35
-
 4-nitrophenyl 2-(acetylamino)-4-O-[2-(acetylamino)-2-deoxy-beta-D-glucopyranosyl]-2-deoxy-beta-D-glucopyranoside pH 6.5, wilde-type recombinant enzyme expressed in Escherichia coli Trichoderma harzianum