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Literature summary extracted from

  • Dennhart, N.; Fukamizo, T.; Brzezinski, R.; Lacombe-Harvey, M.E.; Letzel, T.
    Oligosaccharide hydrolysis by chitosanase enzymes monitored by real-time electrospray ionization-mass spectrometry (2008), J. Biotechnol., 134, 253-260.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.132 wild-type and mutants subcloned into vector pFD666 and expressed in Streptomyces lividans TK-24 Streptomyces sp. N174
3.2.1.165 Streptomyces lividans TK-24 with shuttle vector pFD666, M14 medium, pH 4.2 Amycolatopsis orientalis

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.132 D201A time-course is almost identical to that obtained by the wild-type Streptomyces sp. N174
3.2.1.132 D40G reduces activity to 2% Streptomyces sp. N174

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.165 additional information Amycolatopsis orientalis Chitosanases hydrolyze beta-1,4-glycosidic linkage of chitosan, a polysaccharide consisting mainly of D-glucosamine with a variable content of N-acetyl-D-glucosamine ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.132 Streptomyces sp. N174
-
-
-
3.2.1.165 Amycolatopsis orientalis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.132 by a two-step procedure involving ion exchange chromatography followed by gel filtration, to purity Streptomyces sp. N174
3.2.1.165 SP-Sepharose ion exchange chromatography followed by hydroxyapatite column chromatography Amycolatopsis orientalis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.165 additional information
-
 real-time MS as high sensitivity method to measure chitosanase activity quickly and with low substrate concentration, simultaneous detection of substrate and product Amycolatopsis orientalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.132 GlcNbeta(1-4)GlcNbeta(1-4)GlcNbeta(1-4)GlcN + H2O
-
 Streptomyces sp. N174 GlcNbeta(1-4)GlcN + GlcNbeta(1-4)GlcNbeta(1-4)GlcN + D-glucosamine
-
 ?
3.2.1.132 GlcNbeta(1-4)GlcNbeta(1-4)GlcNbeta(1-4)GlcNbeta(1-4)GlcN + H2O
-
 Streptomyces sp. N174 GlcNbeta(1-4)GlcNbeta(1-4)GlcN + GlcNbeta(1-4)GlcN
-
 ?
3.2.1.132 GlcNbeta(1-4)GlcNbeta(1-4)GlcNbeta(1-4)GlcNbeta(1-4)GlcNbeta(1-4)GlcN + H2O
-
 Streptomyces sp. N174 GlcNbeta(1-4)GlcNbeta(1-4)GlcN + GlcNbeta(1-4)GlcN + GlcNbeta(1-4)GlcNbeta(1-4)GlcNbeta(1-4)GlcN
-
 ?
3.2.1.132 hexa-N-acetylchitohexaose + H2O
-
 Streptomyces sp. N174 ?
-
 ?
3.2.1.132 additional information does not hydrolyze GlcNbeta(1-4)GlcN or GlcNbeta(1-4)GlcNbeta(1-4)GlcN Streptomyces sp. N174 ?
-
 ?
3.2.1.165 chitohexaose + H2O GlcNn: polymerization degree of n=2-6, chitosan oligosaccharides, 0.96 nM exochiosanase, 10 mM ammonium acetate, pH 5.2, substrate concentrations: 6.25, 12.5, 25, 37.5 microM, hexa-N-acetylchitohexaose as control substrate at 25 microM Amycolatopsis orientalis 2-amino-2-deoxy-D-glucopyranose + chitopentaose
-
 ?
3.2.1.165 additional information Chitosanases hydrolyze beta-1,4-glycosidic linkage of chitosan, a polysaccharide consisting mainly of D-glucosamine with a variable content of N-acetyl-D-glucosamine Amycolatopsis orientalis ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.132 endochitosanase
-
 Streptomyces sp. N174
3.2.1.165 exo-beta-glucosaminidase
-
 Amycolatopsis orientalis
3.2.1.165 exochitosanase
-
 Amycolatopsis orientalis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.165 27
-
 chitohexaose 2-amino-2-deoxy-D-glucopyranose split off, pH 5.2, 20°C Amycolatopsis orientalis