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Literature summary for 3.5.1.4 extracted from

  • Nawaz, S.M.; Khan, A.A.; Seng J.E.; Leakey, J.E.; Siitonen, P.H.; Cerniglia, C.E.
    Purification and characterization of an amidase from an acrylamide-degrading Rhodococcus sp. (1994), Appl. Environ. Microbiol., 60, 3343-3348.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
synthesis
-
Brevibacterium sp.
synthesis degradation of acrylamide Rhodococcus sp.

Inhibitors

Inhibitors Comment Organism Structure
aromatic amide
-
Rhodococcus sp.
Co2+
-
Rhodococcus sp.
Cu2+
-
Rhodococcus sp.
Fe2+ activity enhanced Rhodococcus sp.
Hg2+
-
Rhodococcus sp.
iodoacetate
-
Pseudomonas aeruginosa
iodoacetate
-
Rhodococcus sp.
Mg2+ partially inhibited Rhodococcus sp.
Ni2+
-
Rhodococcus sp.
specific iron chelator
-
Rhodococcus sp.
thiol blocking reagent
-
Rhodococcus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2
-
Acrylamide
-
Rhodococcus sp.
2.6
-
Propionamide
-
Rhodococcus sp.
2.7
-
acetamide
-
Rhodococcus sp.
3.2
-
butyramide
-
Rhodococcus sp.
3.5
-
Isobutyramide
-
Rhodococcus sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+ activity enhanced Rhodococcus sp.
Cr2+ activity enhanced Rhodococcus sp.
Iron 8 mol of iron per mol of enzyme, detection by inductively coupled plasma-atomic emission spectrometry Rhodococcus sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
x * 26000, SDS-PAGE Pseudomonas sp.
35000
-
6 * 35000, SDS-PAGE Pseudomonas aeruginosa
43000
-
gel filtration Pseudomonas sp.
43000
-
4 * 43000 Brevibacterium sp.
44500
-
8 * 44500, SDS-PAGE Rhodococcus sp.
180000
-
-
Brevibacterium sp.
200000
-
gel filtration and sedimentation equilibrium analysis, sedimentation coeffient 10.6 S Pseudomonas aeruginosa
360000
-
PAGE, nondenaturing conditions Rhodococcus sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
acrylamide + H2O Rhodococcus sp.
-
acrylic acid + NH3
-
?
butyramide + H2O Rhodococcus sp.
-
n-butyrate + NH3
-
?
isobutyramide + H2O Rhodococcus sp.
-
isobutyrate + NH3
-
?

Organism

Organism UniProt Comment Textmining
Brevibacterium sp.
-
-
-
Pseudomonas aeruginosa
-
-
-
Pseudomonas sp.
-
-
-
Rhodococcus sp.
-
-
-

Purification (Commentary)

Purification (Comment) Organism
acetone precipitation, ammonium sulfate fractionation, DEAE-Sepharose chromatography, Mono Q column and gel filtration, purified approximately 138fold with a yield of 20% Rhodococcus sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
37.6
-
micromol of NH3 * min-1 * mg of protein-1 Pseudomonas sp.
274.1
-
micromol of NH3 * min-1 * mg of protein-1 Brevibacterium sp.
3500
-
micromol of NH3 * min-1 * mg of protein-1 Rhodococcus sp.
5000
-
substrate isobutyramide Rhodococcus sp.
5900
-
substrate acrylamide Rhodococcus sp.
7700
-
substrate butyramide Rhodococcus sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acrylamide + H2O
-
Rhodococcus sp. acrylic acid + NH3
-
?
butyramide + H2O
-
Rhodococcus sp. n-butyrate + NH3
-
?
isobutyramide + H2O
-
Rhodococcus sp. isobutyrate + NH3
-
?

Subunits

Subunits Comment Organism
? x * 26000, SDS-PAGE Pseudomonas sp.
hexamer 6 * 35000, SDS-PAGE Pseudomonas aeruginosa
octamer 8 * 44500, SDS-PAGE Rhodococcus sp.
tetramer 4 * 43000 Brevibacterium sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
at 40°C Rhodococcus sp.

pH Range

pH Minimum pH Maximum Comment Organism
5.5 9
-
Rhodococcus sp.