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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.5.1.42 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.42 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
1.5.1.42 0.00443
-
 FMN recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653
1.5.1.42 0.05166
-
 NADH recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.42 Nocardioides sp. PD653 A0A1V1W312
-
-

Purification (Commentary)

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

Reaction

EC Number Reaction Comment Organism Reaction ID
1.5.1.42 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.42 FMN + NADH + H+
-
 Nocardioides sp. PD653 FMNH2 + NAD+
-
 ?
1.5.1.42 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

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.42 20
-
 assay at Nocardioides sp. PD653

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.42 66.74
-
 NADH recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653
1.5.1.42 66.74
-
 FMN recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.42 7.5
-
 recombinant His-tagged enzyme Nocardioides sp. PD653

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.5.1.42 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

EC Number Cofactor Comment Organism Structure
1.5.1.42 NADH
-
 Nocardioides sp. PD653

General Information

EC Number General Information Comment Organism
1.5.1.42 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.5.1.42 1291.9
-
 NADH recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653
1.5.1.42 15065.5
-
 FMN recombinant enzyme HcbA3, pH 7.5, 20°C Nocardioides sp. PD653