Taiwanofungus camphorata nitroreductase

CDNA cloning and biochemical characterisation

Chih Chen Chen, Chuian-Fu Ken, Lisa Wen, Ching Fong Chang, Chi Tsai Lin

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Nitroreductases (Nrs) play important roles in redox system via NADPH or NADH as a reductant. A TcNr cDNA encoding a putative Nr was cloned from Taiwanofungus camphorata. A 3-D structural model of the TcNr has been created based on the known structure of BcNr (Bacillus cereus). To characterise the TcNr, the coding region was subcloned into an expression vector and transformed into Escherichia coli. The recombinant His6-tagged TcNr was purified by Ni affinity chromatography. The purified enzyme showed a single band at molecular mass of approximately 25 kDa on 12% sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The enzyme exhibited Nr activity via ferricyanide assay. The Michaelis constant (KM) value for ferricyanide was 0.86 mM. The enzyme's half-life of deactivation at 45 °C was 12.3 min. The enzyme was most active at pH 6. The enzyme's preferred substrate is 1-chloro-2, 4-dinitrobenzene.

Original languageEnglish
Pages (from-to)2708-2713
Number of pages6
JournalFood Chemistry
Volume135
Issue number4
DOIs
Publication statusPublished - 2012 Dec 15

Fingerprint

Taiwanofungus
Nitroreductases
Cloning
Organism Cloning
Enzymes
enzymes
ferricyanides
Bacillus cereus
Affinity chromatography
Dinitrochlorobenzene
reducing agents
NAD (coenzyme)
Structural Models
Reducing Agents
Molecular mass
affinity chromatography
Electrophoresis
NADP
Affinity Chromatography
NADP (coenzyme)

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Food Science

Cite this

Chen, Chih Chen ; Ken, Chuian-Fu ; Wen, Lisa ; Chang, Ching Fong ; Lin, Chi Tsai. / Taiwanofungus camphorata nitroreductase : CDNA cloning and biochemical characterisation. In: Food Chemistry. 2012 ; Vol. 135, No. 4. pp. 2708-2713.
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abstract = "Nitroreductases (Nrs) play important roles in redox system via NADPH or NADH as a reductant. A TcNr cDNA encoding a putative Nr was cloned from Taiwanofungus camphorata. A 3-D structural model of the TcNr has been created based on the known structure of BcNr (Bacillus cereus). To characterise the TcNr, the coding region was subcloned into an expression vector and transformed into Escherichia coli. The recombinant His6-tagged TcNr was purified by Ni affinity chromatography. The purified enzyme showed a single band at molecular mass of approximately 25 kDa on 12{\%} sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The enzyme exhibited Nr activity via ferricyanide assay. The Michaelis constant (KM) value for ferricyanide was 0.86 mM. The enzyme's half-life of deactivation at 45 °C was 12.3 min. The enzyme was most active at pH 6. The enzyme's preferred substrate is 1-chloro-2, 4-dinitrobenzene.",
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Taiwanofungus camphorata nitroreductase : CDNA cloning and biochemical characterisation. / Chen, Chih Chen; Ken, Chuian-Fu; Wen, Lisa; Chang, Ching Fong; Lin, Chi Tsai.

In: Food Chemistry, Vol. 135, No. 4, 15.12.2012, p. 2708-2713.

Research output: Contribution to journalArticle

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T2 - CDNA cloning and biochemical characterisation

AU - Chen, Chih Chen

AU - Ken, Chuian-Fu

AU - Wen, Lisa

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