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Literature summary for 1.5.1.42 extracted from

  • Ito, K.; Takagi, K.; Kataoka, R.; Kiyota, H.
    Biochemical characterization of NADH FMN oxidoreductase HcbA3 from Nocardioides sp. PD653 in catalyzing aerobic HCB dechlorination (2020), J. Pestic. Sci., 45, 125-131 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.5.1.42

Cloned(Commentary)

Cloned (Comment) Organism
gene hcbA3, recombinant expression of C-terminally His-tagged enzyme in Rhodococcus erythropolis, subcloning in Escherichia coli strain DH5alpha Nocardioides sp. PD653

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information steady-state kinetic analysis, the initial rates of NADH oxidation follow typical Michaelis-Menten kinetics, and Lineweaver-Burk plots show parallel patterns for a ping-pong mechanism Nocardioides sp. PD653
0.00443
-
 FMN recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653
0.05166
-
 NADH recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
FMN + NADH + H+ Nocardioides sp. PD653
-
 FMNH2 + NAD+
-
 ?

Organism

Organism UniProt Comment Textmining
Nocardioides sp. PD653 A0A1V1W312
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant C-terminally His-tagged enzyme from Rhodococcus erythropolis by metal affinity chromatography Nocardioides sp. PD653

Reaction

Reaction Comment Organism Reaction ID
FMNH2 + NAD+ = FMN + NADH + H+ HcbA3 follows a ping-pong bi-bi mechanism for the reduction of flavins Nocardioides sp. PD653
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FMN + NADH + H+
-
 Nocardioides sp. PD653 FMNH2 + NAD+
-
 ?
additional information HcbA3 is an NADH:FMN oxidoreductase. Successful reconstitution of the oxidative dehalogenase reaction in vitro, which consists of HcbA1, HcbA3, FMN, and NADH, suggesting that HcbA3 may also be the partner reductase component for HcbA1 in Nocardioides sp. PD653. The optimal molar ratio of HcbA1 and HcbA3 is 7:1 Nocardioides sp. PD653 ?
-
-

Synonyms

Synonyms Comment Organism
HcbA3
-
 Nocardioides sp. PD653
hexachlorobenzene oxidative dehalogenase system reductase component UniProt Nocardioides sp. PD653
NADH:FMN oxidoreductase
-
 Nocardioides sp. PD653

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
20
-
 assay at Nocardioides sp. PD653

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
66.74
-
 NADH recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653
66.74
-
 FMN recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 recombinant His-tagged enzyme Nocardioides sp. PD653

pH Range

pH Minimum pH Maximum Comment Organism
7 8.6 at pH 7.0, 7.2, and 8.0, the enzyme activity levels in 100 mM phosphate buffer are approx. 50.2, 88.1, and 81.1%, respectively, relative to that at pH 7.5. HcbA3C-His shows the highest flavin reductase activity in 60 mM KPi buffer Nocardioides sp. PD653

Cofactor

Cofactor Comment Organism Structure
NADH
-
 Nocardioides sp. PD653

General Information

General Information Comment Organism
physiological function Nocardioides sp. PD653 genes hcbA1, hcbA2, and hcbA3 encode enzymes that catalyze the oxidative dehalogenation of hexachlorobenzene (HCB), which is one of the most recalcitrant persistent organic pollutants (POPs). HcbA3 shows the highest affinity for FMN. HcbA3 may be the partner reductase component for HcbA1 in Nocardioides sp. PD653 Nocardioides sp. PD653

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1291.9
-
 NADH recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653
15065.5
-
 FMN recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653