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Literature summary for 3.5.1.105 extracted from

  • Jiang, Z.; Niu, T.; Lv, X.; Liu, Y.; Li, J.; Lu, W.; Du, G.; Chen, J.; Liu, L.
    Secretory expression fine-tuning and directed evolution of diacetylchitobiose deacetylase by Bacillus subtilis (2019), Appl. Environ. Microbiol., 85, e01076-19 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.1.105

Application

Application Comment Organism
synthesis Dac is a key enzyme used in the biodegradation of chitin and chitosan to produce chitosan oligosaccharides and monosaccharides Pyrococcus horikoshii

Cloned(Commentary)

Cloned (Comment) Organism
gene PH0499, recombinant expression of wild-type and mutants in Bacillus subtilis strain WB600 under control of the HpaII promoter and secreteion as extracellular enzymes, screening of signal peptides from Bacillus subtilis for optimal expression. The signal peptide YncM achieves the highest extracellular diacetylchitobiose deacetylase activity of 13.5 U/ml. Subcloning in Escherichia coli. Quantitative real-time PCR expression analysis. Method optimization, overview Pyrococcus horikoshii

Protein Variants

Protein Variants Comment Organism
R157H site-directed mutagenesis, the mutant exhibits increased specific activity compared to the wild-type Pyrococcus horikoshii
R157T site-directed mutagenesis, the mutant R157T exhibits much higher specific activity than the wild-type. Achievement of efficient secretory production and improvement of the catalytic efficiency of diacetylchitobiose deacetylase in Bacillus subtilis Pyrococcus horikoshii
R157W site-directed mutagenesis, the mutant exhibits increased specific activity compared to the wild-type Pyrococcus horikoshii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.19
-
 N,N'-diacetylchitobiose pH 8.0, 37°C, mutant R157T Pyrococcus horikoshii
5.34
-
 N,N'-diacetylchitobiose pH 8.0, 37°C, mutant R157H Pyrococcus horikoshii
5.39
-
 N,N'-diacetylchitobiose pH 8.0, 37°C, mutant R157W Pyrococcus horikoshii
7.04
-
 N,N'-diacetylchitobiose pH 8.0, 37°C, wild-type enzyme Pyrococcus horikoshii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N,N'-diacetylchitobiose + H2O Pyrococcus horikoshii
-
 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O Pyrococcus horikoshii DSM 12428
-
 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O Pyrococcus horikoshii NBRC 100139
-
 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O Pyrococcus horikoshii JCM 9974
-
 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O Pyrococcus horikoshii ATCC 700860
-
 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O Pyrococcus horikoshii OT-3
-
 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?

Organism

Organism UniProt Comment Textmining
Pyrococcus horikoshii O58235
-
-
Pyrococcus horikoshii ATCC 700860 O58235
-
-
Pyrococcus horikoshii DSM 12428 O58235
-
-
Pyrococcus horikoshii JCM 9974 O58235
-
-
Pyrococcus horikoshii NBRC 100139 O58235
-
-
Pyrococcus horikoshii OT-3 O58235
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant engineered extracellular enzyme mutant enzymes from Bacillus subtilis by anion exchange chromatography Pyrococcus horikoshii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2047.3
-
 pH 8.0, 37°C, purified wild-type enzyme Pyrococcus horikoshii
3112.2
-
 pH 8.0, 37°C, purified mutant R157T Pyrococcus horikoshii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N,N'-diacetylchitobiose + H2O
-
 Pyrococcus horikoshii N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O
-
 Pyrococcus horikoshii DSM 12428 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O
-
 Pyrococcus horikoshii NBRC 100139 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O
-
 Pyrococcus horikoshii JCM 9974 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O
-
 Pyrococcus horikoshii ATCC 700860 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?
N,N'-diacetylchitobiose + H2O
-
 Pyrococcus horikoshii OT-3 N-acetyl-beta-D-glucosaminyl-(1->4)-D-glucosamine + acetate
-
 ?

Subunits

Subunits Comment Organism
? x * 32000, recombinant enzyme, SDS-PAGE Pyrococcus horikoshii

Synonyms

Synonyms Comment Organism
Dac
-
 Pyrococcus horikoshii
diacetylchitobiose deacetylase
-
 Pyrococcus horikoshii
PH0499
-
 Pyrococcus horikoshii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Pyrococcus horikoshii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
85
-
 purified recombinant enzyme, 30 min, stable Pyrococcus horikoshii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Pyrococcus horikoshii

General Information

General Information Comment Organism
evolution the enzyme belongs to the carbohydrate esterase family 14 Pyrococcus horikoshii
metabolism diacetylchitobiose deacetylase works in the chitin degradation pathway in combination with glucosaminidase to hydrolyze diacetylchitobiose (GlcNAc2) to glucosamine (GlcN). First, the N-acetyl group of GlcNAc2 is catalyzed by Dac from the nonreducing end residue N-acetylglucosamine (GlcNAc) of GlcNAc2 and generates the product GlcN-GlcNAc, the product (GlcNGlcNAc) is then hydrolyzed by glucosaminidase following degradation into GlcN and GlcNAc. Finally, the resulting monomer GlcNAc is catalyzed by Dac to generate GlcN Pyrococcus horikoshii
physiological function the diacetylchitobiose deacetylase (Dac) plays an important role in a unique chitin degradation pathway in Archaea Pyrococcus horikoshii