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Literature summary extracted from

  • Liao, S.; Poonpairoj, P.; Ko, K.; Takatuska, Y.; Yamaguchi, Y.; Abe, N.; Kaneko, J.; Kamio, Y.
    Occurrence of agmatine pathway for putrescine synthesis in Selenomonas ruminatium (2008), Biosci. Biotechnol. Biochem., 72, 445-455.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.53 expressed in Escherichia coli Selenomonas ruminantium
3.5.1.53 expression in Escherichia coli strain DE3 Selenomonas ruminantium
3.5.3.12 expression in Escherichia coli strain DE3 Selenomonas ruminantium
4.1.1.19 expression in Escherichia coli strain DE3 Selenomonas ruminantium

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.53 C153A inactive mutant enzyme Selenomonas ruminantium
3.5.1.53 C153A mutant without enzyme activity Selenomonas ruminantium
3.5.1.53 E160A inactive mutant enzyme Selenomonas ruminantium
3.5.1.53 E160A mutant without enzyme activity Selenomonas ruminantium
3.5.3.12 C361A 20% enzyme activity of wild-type enzyme Selenomonas ruminantium

General Stability

EC Number General Stability Organism
3.5.1.53 in 50 mM PIPES buffer at pH 6.5 stable for 30 min up to 70°C Selenomonas ruminantium
4.1.1.19 at 50°C in absence of PALP lost of 90% enzyme activity within 10 min, no enzyme activity after heating at 70°C for 60 min in presence of PALP Selenomonas ruminantium

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.53 Co2+ strong inhibitor Selenomonas ruminantium
3.5.1.53 Fe2+ strong inhibitor Selenomonas ruminantium
3.5.1.53 Hg2+ strong inhibitor Selenomonas ruminantium
3.5.1.53 iodoacetamide strong inhibitor Selenomonas ruminantium
3.5.1.53 Mn2+ strong inhibitor Selenomonas ruminantium
3.5.1.53 Zn2+ strong inhibitor Selenomonas ruminantium
3.5.3.12 Al3+ 1 mM Selenomonas ruminantium
3.5.3.12 Co2+ 1 mM Selenomonas ruminantium
3.5.3.12 Cu2+ 1 mM Selenomonas ruminantium
3.5.3.12 Fe2+ 1 mM Selenomonas ruminantium
3.5.3.12 Fe3+ 1 mM Selenomonas ruminantium
3.5.3.12 Hg2+ strong inhibitor Selenomonas ruminantium
3.5.3.12 iodoacetamide strong inhibitor Selenomonas ruminantium
3.5.3.12 Ni2+ 1 mM Selenomonas ruminantium
3.5.3.12 Zn2+ 1 mM Selenomonas ruminantium
4.1.1.19 DL-alpha-difluoromethylarginine
-
 Selenomonas ruminantium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.53 0.22
-
 N-Carbamoylputrescine
-
 Selenomonas ruminantium
3.5.3.12 6.67
-
 agmatine
-
 Selenomonas ruminantium
4.1.1.19 5.6
-
 L-arginine
-
 Selenomonas ruminantium
4.1.1.19 50
-
 L-lysine
-
 Selenomonas ruminantium

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.53 32532
-
 2 * 32532, calculated from sequence Selenomonas ruminantium
3.5.1.53 37000
-
 SDS-PAGE Selenomonas ruminantium
3.5.1.53 37000
-
 2 * 37000, SDS-PAGE Selenomonas ruminantium
3.5.1.53 80000
-
 gel filtration Selenomonas ruminantium
3.5.3.12 45000
-
 gel filtration Selenomonas ruminantium
3.5.3.12 45000
-
 SDS-PAGE Selenomonas ruminantium
4.1.1.19 58000
-
 SDS-PAGE Selenomonas ruminantium
4.1.1.19 120000
-
 gel filtration Selenomonas ruminantium

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.53 Selenomonas ruminantium
-
-
-
3.5.1.53 Selenomonas ruminantium Q5KR04
-
-
3.5.3.12 Selenomonas ruminantium Q5KR05
-
-
4.1.1.19 Selenomonas ruminantium Q5NT93
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.53
-
 Selenomonas ruminantium
3.5.1.53 recombinant Selenomonas ruminantium
3.5.3.12
-
 Selenomonas ruminantium
4.1.1.19 2024fold after final purification step with MonoQ HR 5/5 Selenomonas ruminantium

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.1.19 0.00738
-
 crude extract Selenomonas ruminantium
4.1.1.19 0.01104
-
 heat treatment Selenomonas ruminantium
4.1.1.19 0.02394
-
 after purification with DEAE-Toyopearl 650M Selenomonas ruminantium
4.1.1.19 0.0381
-
 after purification with DEAE-5PW Selenomonas ruminantium
4.1.1.19 1.026
-
 at the semi-final step of purification with HA-1000 column chromatography Selenomonas ruminantium
4.1.1.19 14.94
-
 final step of purification with MonoQ HR 5/5 Selenomonas ruminantium

Storage Stability

EC Number Storage Stability Organism
4.1.1.19 4°C, 1 month, no lost of enzyme activity, pH 6.0-7.5 Selenomonas ruminantium
4.1.1.19 50°C, absence of PALP, 10 min, only 10% residual activity Selenomonas ruminantium
4.1.1.19 70°C, presence of PALP, 60 min, no residual activity Selenomonas ruminantium

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.53 N-carbamoylputrescine + H2O
-
 Selenomonas ruminantium putrescine + CO2 + NH3
-
 ?
3.5.1.53 N-carbamoylputrescine + H2O 10 mM, assay at pH 6.5, 10 min, 40°C Selenomonas ruminantium putrescine + CO2 + NH3
-
 ?
3.5.3.12 agmatine + H2O 25 mM, assay at pH 6.5, 10 min, 40°C Selenomonas ruminantium putrescine + ?
-
 ?
4.1.1.19 L-arginine assay at pH 6.5, 10 min, 50°C Selenomonas ruminantium agmatine + CO2
-
 ?
4.1.1.19 L-lysine assay at pH 6.5, 10 min, 50°C Selenomonas ruminantium ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.5.1.53 dimer 2 * 37000, SDS-PAGE Selenomonas ruminantium
3.5.1.53 dimer 2 * 32532, calculated from sequence Selenomonas ruminantium
3.5.1.53 homodimer
-
 Selenomonas ruminantium
3.5.3.12 monomer
-
 Selenomonas ruminantium
4.1.1.19 dimer
-
 Selenomonas ruminantium

Synonyms

EC Number Synonyms Comment Organism
3.5.1.53 AguB
-
 Selenomonas ruminantium
3.5.1.53 N-carbamoyl-putrescine amidohydrolase
-
 Selenomonas ruminantium
3.5.1.53 N-carbamoylputrescine amidohydrolase
-
 Selenomonas ruminantium
3.5.1.53 NCP amidohydrolase
-
 Selenomonas ruminantium
3.5.3.12 agmatine deiminase
-
 Selenomonas ruminantium
3.5.3.12 AguA
-
 Selenomonas ruminantium
4.1.1.19 ADC
-
 Selenomonas ruminantium
4.1.1.19 arginine decarboxylase
-
 Selenomonas ruminantium
4.1.1.19 L-Arginine decarboxylase
-
 Selenomonas ruminantium

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.53 45
-
-
 Selenomonas ruminantium
3.5.1.53 45
-
 assay at pH 6.5, 10 min, 40°C Selenomonas ruminantium
3.5.3.12 40
-
 assay at Selenomonas ruminantium
4.1.1.19 60
-
-
 Selenomonas ruminantium

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.1.1.19 40 70 after 10 min incubation at 50°C 4fold higher enzyme activity than at 40°C, after 10 min incubation at 55°C 7fold higher enzyme activity than at 40°C, after 10 min incubation at 60°C 10fold higher enzyme activity than at 40°C, after 10 min incubation at 70°C 1.2fold higher enzyme activity than at 40°C Selenomonas ruminantium

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.1.53 70
-
 AguB activity is stable at up to 70°C for 30 min in 50 mM PIPES buffer, pH 6.5 Selenomonas ruminantium
3.5.1.53 70
-
 up to 70°C, stable for 30 min at pH 6.5, 50 mM PIPES buffer Selenomonas ruminantium
3.5.3.12 40
-
 below 40°C stable for 30 min, pH 6.0-9.0 Selenomonas ruminantium
4.1.1.19 35
-
 without pyridoxal 5'-phosphate, 50% redsidual enzyme activity Selenomonas ruminantium
4.1.1.19 60
-
 with pyridoxal 5'-phosphate, 54% residual enzyme activity Selenomonas ruminantium

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.53 7
-
-
 Selenomonas ruminantium
3.5.1.53 7
-
 assay at pH 6.5, 10 min, 40°C Selenomonas ruminantium
3.5.3.12 6.5
-
 assay at Selenomonas ruminantium
4.1.1.19 6.5
-
 assay at pH 6.5, 10 min, 50°C Selenomonas ruminantium

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.1.1.19 6.5 9 80% enzyme activity at pH 7.0, 70% enzyme activity at pH 8.0, 50% enzyme activity at pH 9.0 Selenomonas ruminantium

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.5.1.53 6 8
-
 Selenomonas ruminantium
3.5.1.53 6 8 stable Selenomonas ruminantium
3.5.3.12 6 9 30 min, temperature below 40°C Selenomonas ruminantium
4.1.1.19 6.5 7.5 more than 80% enzyme activity, decreased enzyme activity at higher pH Selenomonas ruminantium

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.1.19 pyridoxal 5'-phosphate
-
 Selenomonas ruminantium

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
4.1.1.19 0.8
-
 DL-alpha-difluoromethylarginine
-
 Selenomonas ruminantium