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Literature summary for 4.2.2.3 extracted from

  • Uchimura, K.; Miyazaki, M.; Nogi, Y.; Kobayashi, T.; Horikoshi, K.
    Cloning and sequencing of alginate lyase genes from deep-sea strains of Vibrio and Agarivorans and characterization of a new Vibrio enzyme (2010), Mar. Biotechnol., 12, 526-533.
    View publication on PubMed
Search for more literature for 4.2.2.3

Activating Compound

Activating Compound Comment Organism Structure
(NH4)2SO4 (NH4)2SO4 at 0.2 M increases the original activity of isoform A9mT 23.5times Vibrio sp.
CH3COO- NH4+ CH3COONH4 at 0.2 M increases the original activity of isoform A9mT 20.9times Vibrio sp.
CH3COONa CH3COONa at 0.2 M increases the original activity of isoform A9mT 21times Vibrio sp.
CsCl CsCl at 0.2 M increases the original activity of isoform A9mT 25times Vibrio sp.
KCl KCl at 0.2 M increases the original activity of isoform A9mT 24.9times Vibrio sp.
LiCl LiCl at 0.2 M increases the original activity of isoform A9mT 18.8times Vibrio sp.
additional information 5 mM EDTA and EGTA do not influence enzyme activity of isoform A9mT Vibrio sp.
NaBr NaBr at 0.2 M increases the original activity of isoform A9mT 21.8times Vibrio sp.
NaCl recombinant isoform A9mT is remarkably activated in the presence of NaCl, when 0.4 M NaCl is added to the reaction mixture, the activity is increased maximally to 24times the original activity, the activity gradually decreases at higher NaCl concentrations, but is still activated 20times at 0.8 M and 8 times at 1.6 M NaCl Vibrio sp.
NaF NaF at 0.2 M increases the original activity of isoform A9mT 21.7times Vibrio sp.
NH4Cl NH4Cl at 0.2 M increases the original activity of isoform A9mT 22.7times Vibrio sp.
Nonidet P40 the enzyme is moderately activated by Nonidet P40 by 134% Vibrio sp.
RbCl RbCl at 0.2 M increases the original activity of isoform A9mT 25times Vibrio sp.
Tween 20 the enzyme is moderately activated by Tween 20 by 139% Vibrio sp.

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli DH5alpha cells Vibrio sp.
expressed in Escherichia coli DH5alpha cells Agarivorans sp.
expression in Escherichia coli Vibrio sp.

Inhibitors

Inhibitors Comment Organism Structure
Co2+ isoform A9mT is inhibited by 14% at 1 mM Vibrio sp.
Cu2+ isoform A9mT is inhibited by 16% at 1 mM Vibrio sp.
NaCl in the presence of 0.2 M NaCl, the residual activities of isoform A9mL from pH 6 to 10 are decreased 40-75% of those observed in the absence of NaCl Vibrio sp.
Ni2+ isoform A9mT is inhibited by 35% at 1 mM Vibrio sp.
SDS strongly inhibits the enzyme activity by 90% Vibrio sp.
Sr2+ isoform A9mT is inhibited by 14% at 1 mM Vibrio sp.
Zn2+ isoform A9mT is inhibited by 19% at 1 mM Vibrio sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Cs+ 0.2 M, 2500% of initial activity Vibrio sp.
K+ 0.2 M, 2500% of initial activity Vibrio sp.
Li+ 0.2 M, 1880% of initial activity Vibrio sp.
additional information Sn2+, Ca2+, Mn2+, and Mg2+ do not influence enzyme activity of isoform A9mT Vibrio sp.
Na+ 0.2 M, 2200% of initial activity Vibrio sp.
NH4+ 0.2 M, 2270% of initial activity Vibrio sp.
Rb+ 0.2 M, 2500% of initial activity Vibrio sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
25010
-
 isoform A9mC, calculated from amino acid sequence Vibrio sp.
28000
-
 isoform A9mT, SDS-PAGE Vibrio sp.
29640
-
 isoform A9mT, calculated from amino acid sequence Vibrio sp.
29643
-
 1 * 29643, calculated, 1 * 28000, SDS-PAGE Vibrio sp.
30218
-
 x * 30218, calculated Agarivorans sp.
30220
-
 isoform A1mU, calculated from amino acid sequence Agarivorans sp.
31000
-
 gel filtration Vibrio sp.
31000
-
 isoform A9mT, Bio-Gel-A0.5 m gel filtration Vibrio sp.
54430
-
 isoform A9mL, calculated from amino acid sequence Vibrio sp.

Organism

Organism UniProt Comment Textmining
Agarivorans sp. C4TIN8
-
-
Agarivorans sp. JAM-A1m C4TIN8
-
-
Vibrio sp.
-
 sequence contains a putative signal sequence of 23 amino acids
-
Vibrio sp. C4TJD2
-
-
Vibrio sp. C4TJD2 isoform A9mT; strain JAM-A9m
-
Vibrio sp. C4TJD3 isoform A9mC; strain JAM-A9m
-
Vibrio sp. C4TJD4 isoform A9mL; strain JAM-A9m
-
Vibrio sp. A9m
-
 sequence contains a putative signal sequence of 23 amino acids
-
Vibrio sp. A9m C4TJD2
-
-
Vibrio sp. A9m C4TJD2 isoform A9mT; strain JAM-A9m
-
Vibrio sp. A9m C4TJD3 isoform A9mC; strain JAM-A9m
-
Vibrio sp. A9m C4TJD4 isoform A9mL; strain JAM-A9m
-

Posttranslational Modification

Posttranslational Modification Comment Organism
proteolytic modification sequence contains a putative signal sequence of 23 amino acids Vibrio sp.

Purification (Commentary)

Purification (Comment) Organism
DEAE-Toyopearl column chromatography Vibrio sp.
DEAE-Toyopearl column chromatography Agarivorans sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1401
-
 pH 7.5, 30°C Vibrio sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alginate preferably degrades M blocks Agarivorans sp. ?
-
 ?
alginate preferably degrades M blocks Agarivorans sp. JAM-A1m ?
-
 ?
alginate
-
 Vibrio sp. unsaturated algino-oligosaccharides
-
 ?
alginate
-
 Agarivorans sp. unsaturated algino-oligosaccharides
-
 ?
alginate isoform A9mT favorably degrades mannuronate polymer in alginate, the relative activities for alginate, beta-D-mannuronic acid, 1,4 linked alpha-L-guluronic acid, and 1,4 linked alpha-L-guluronic acid/beta-D-mannuronic acid blocks are 100%, 75%, 21%, and 15%, respectively, under the standard assay conditions at pH 7.5 Vibrio sp. unsaturated algino-oligosaccharides
-
 ?
alginate isoform A9mT favorably degrades mannuronate polymer in alginate, the relative activities for alginate, beta-D-mannuronic acid, 1,4 linked alpha-L-guluronic acid, and 1,4 linked alpha-L-guluronic acid/beta-D-mannuronic acid blocks are 100%, 75%, 21%, and 15%, respectively, under the standard assay conditions at pH 7.5 Vibrio sp. A9m unsaturated algino-oligosaccharides
-
 ?
alginate
-
 Vibrio sp. A9m unsaturated algino-oligosaccharides
-
 ?
alginate
-
 Agarivorans sp. JAM-A1m unsaturated algino-oligosaccharides
-
 ?
beta-D-mannuronyl linkage in alginate preferably degrades the M block over the G block in alginate Vibrio sp. ?
-
 ?
beta-D-mannuronyl linkage in alginate preferably degrades the M block over the G block in alginate Vibrio sp. A9m ?
-
 ?
additional information the relative activities for alginate, M, G, and GM blocks are 100%, 75%, 21%, and 15%, respectively Vibrio sp. ?
-
 ?
additional information the relative activities for alginate, M, G, and GM blocks are 100%, 75%, 21%, and 15%, respectively Vibrio sp. A9m ?
-
 ?
poly(beta-D-mannuronate)
-
 Agarivorans sp. ?
-
 ?
poly(beta-D-mannuronate)
-
 Agarivorans sp. JAM-A1m ?
-
 ?

Subunits

Subunits Comment Organism
? x * 30218, calculated Agarivorans sp.
monomer 1 * 31000, isoform A9mT, Bio-Gel-A0.5 m gel filtration Vibrio sp.
monomer 1 * 29643, calculated, 1 * 28000, SDS-PAGE Vibrio sp.

Synonyms

Synonyms Comment Organism
A1mU isoform Agarivorans sp.
A9mC isoform Vibrio sp.
A9mL isoform Vibrio sp.
A9mT
-
 Vibrio sp.
A9mT isoform Vibrio sp.
alg
-
 Agarivorans sp.
alginate lyase
-
 Vibrio sp.
alginate lyase
-
 Agarivorans sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 recombinant isoform A9mT, around 30°C in the presence of 0.2 M NaCl at pH 7.5 Vibrio sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
5 50 the relative activities of isoform A9mT at 10°C and 2°C are around 45% and 30% of the maximal activity at 30°C, respectively, isoform A9mT shows the highest thermal stability in the presence of NaCl, recombinant isoform A9mT is rapidly inactivated above 30°C when the enzyme is incubated at 5°C to 50°C for 15 min in 100 mM Tris-HCl buffer (pH 7.5) with or without 0.2 M NaCl and the enzyme is rapidly inactivated when incubated at above 40°C in the absence of NaCl Vibrio sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6 9
-
 Vibrio sp.
7.6 9 recombinant isoform A9mT, in 100 mM Tris-HCl buffer, MOPS buffer, and glycine-NaOH buffer in the presence of 0.2 M NaCl Vibrio sp.
8 9 crude recombinant A9mL isozyme Vibrio sp.

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 10 isoform A9mT is stable at pH 7.0-10.0 in various 20 mM buffers without NaCl after incubation at 30°C for 15 min Vibrio sp.
9.5 10 the relative activities at pH 9.5 and 10 in glycine-NaOH buffer are approximately 50% and 20%, respectively, of the maximal activity of isoform A9mL Vibrio sp.

pI Value

Organism Comment pI Value Maximum pI Value
Vibrio sp. isoform A9mC, calculated from amino acid sequence
-
 4.7
Vibrio sp. isoform A9mL, calculated from amino acid sequence
-
 4.9
Agarivorans sp. calculated
-
 6.4
Agarivorans sp. isoform A1mU, calculated from amino acid sequence
-
 6.4
Vibrio sp. calculated
-
 7.2
Vibrio sp. isoform A9mT, calculated from amino acid sequence
-
 7.2