Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase

Akito Nishizawa, Ayaka Harada, Miki Senda, Yuka Tachihara, Daisuke Muramatsu, Shinya Kishigami, Shigemasa Mori, Keisuke Sugiyama, Toshiya Senda, Shigenobu Kimura

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The coenzyme specificity of enzymes is one of the critical parameters for the engineered production of biological compounds using bacteria. SinceNADPH is produced abundantly in photosynthetic organisms, conversion of an NADH-specific enzyme into an NADPH-specific one is a useful approach for the efficient carbon-neutral production of biological compounds in photosynthetic organisms. In the present study, anNADH-specific ferredoxin reductase component, BphA4 of biphenyl dioxygenase BphA from Acidovorax sp. Strain KKS102, was changed to an NADPH-dependent form using a method combining structure-based systematic mutations and site-directed random mutagenesis. The resultant CRG mutant, in which Glu175-Thr176-Gln177 of an NADH-recognition loop in the wild-type BphA4 was replaced with Cys 175-Arg176-Gly177, was highly specific and active for NADPH, and its biochemical and structural properties for NADPH were nearly the same as those of the wild-type BphA4 for NADH. In addition, this mutation project was assessed by a semiempirical prediction method of mutation effects, and the results suggested that the CRG mutant was one of the best NADPHspecific mutants.

Original languageEnglish
Pages (from-to)257-265
Number of pages9
JournalBiochemical Journal
Volume462
Issue number2
DOIs
Publication statusPublished - 2014 Sep 1
Externally publishedYes

Fingerprint

Ferredoxins
NADP
NAD
Oxidoreductases
Nucleotides
Mutation
Comamonadaceae
Enzymes and Coenzymes
Dioxygenases
Mutagenesis
Coenzymes
Enzymes
Site-Directed Mutagenesis
Structural properties
Bacteria
Carbon
pyridine

Keywords

  • Biphenyl degradation
  • Electron transfer
  • Flavoprotein
  • Protein design
  • Random mutagenesis
  • Specificity prediction

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Nishizawa, A., Harada, A., Senda, M., Tachihara, Y., Muramatsu, D., Kishigami, S., ... Kimura, S. (2014). Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase. Biochemical Journal, 462(2), 257-265. https://doi.org/10.1042/BJ20140384

Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase. / Nishizawa, Akito; Harada, Ayaka; Senda, Miki; Tachihara, Yuka; Muramatsu, Daisuke; Kishigami, Shinya; Mori, Shigemasa; Sugiyama, Keisuke; Senda, Toshiya; Kimura, Shigenobu.

In: Biochemical Journal, Vol. 462, No. 2, 01.09.2014, p. 257-265.

Research output: Contribution to journalArticle

Nishizawa, A, Harada, A, Senda, M, Tachihara, Y, Muramatsu, D, Kishigami, S, Mori, S, Sugiyama, K, Senda, T & Kimura, S 2014, 'Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase', Biochemical Journal, vol. 462, no. 2, pp. 257-265. https://doi.org/10.1042/BJ20140384
Nishizawa, Akito ; Harada, Ayaka ; Senda, Miki ; Tachihara, Yuka ; Muramatsu, Daisuke ; Kishigami, Shinya ; Mori, Shigemasa ; Sugiyama, Keisuke ; Senda, Toshiya ; Kimura, Shigenobu. / Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase. In: Biochemical Journal. 2014 ; Vol. 462, No. 2. pp. 257-265.
@article{a1ff7982d9394dfd8972b6897b8ea3cf,
title = "Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase",
abstract = "The coenzyme specificity of enzymes is one of the critical parameters for the engineered production of biological compounds using bacteria. SinceNADPH is produced abundantly in photosynthetic organisms, conversion of an NADH-specific enzyme into an NADPH-specific one is a useful approach for the efficient carbon-neutral production of biological compounds in photosynthetic organisms. In the present study, anNADH-specific ferredoxin reductase component, BphA4 of biphenyl dioxygenase BphA from Acidovorax sp. Strain KKS102, was changed to an NADPH-dependent form using a method combining structure-based systematic mutations and site-directed random mutagenesis. The resultant CRG mutant, in which Glu175-Thr176-Gln177 of an NADH-recognition loop in the wild-type BphA4 was replaced with Cys 175-Arg176-Gly177, was highly specific and active for NADPH, and its biochemical and structural properties for NADPH were nearly the same as those of the wild-type BphA4 for NADH. In addition, this mutation project was assessed by a semiempirical prediction method of mutation effects, and the results suggested that the CRG mutant was one of the best NADPHspecific mutants.",
keywords = "Biphenyl degradation, Electron transfer, Flavoprotein, Protein design, Random mutagenesis, Specificity prediction",
author = "Akito Nishizawa and Ayaka Harada and Miki Senda and Yuka Tachihara and Daisuke Muramatsu and Shinya Kishigami and Shigemasa Mori and Keisuke Sugiyama and Toshiya Senda and Shigenobu Kimura",
year = "2014",
month = "9",
day = "1",
doi = "10.1042/BJ20140384",
language = "English",
volume = "462",
pages = "257--265",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - Complete pyridine-nucleotide-specific conversion of an NADH-dependent ferredoxin reductase

AU - Nishizawa, Akito

AU - Harada, Ayaka

AU - Senda, Miki

AU - Tachihara, Yuka

AU - Muramatsu, Daisuke

AU - Kishigami, Shinya

AU - Mori, Shigemasa

AU - Sugiyama, Keisuke

AU - Senda, Toshiya

AU - Kimura, Shigenobu

PY - 2014/9/1

Y1 - 2014/9/1

N2 - The coenzyme specificity of enzymes is one of the critical parameters for the engineered production of biological compounds using bacteria. SinceNADPH is produced abundantly in photosynthetic organisms, conversion of an NADH-specific enzyme into an NADPH-specific one is a useful approach for the efficient carbon-neutral production of biological compounds in photosynthetic organisms. In the present study, anNADH-specific ferredoxin reductase component, BphA4 of biphenyl dioxygenase BphA from Acidovorax sp. Strain KKS102, was changed to an NADPH-dependent form using a method combining structure-based systematic mutations and site-directed random mutagenesis. The resultant CRG mutant, in which Glu175-Thr176-Gln177 of an NADH-recognition loop in the wild-type BphA4 was replaced with Cys 175-Arg176-Gly177, was highly specific and active for NADPH, and its biochemical and structural properties for NADPH were nearly the same as those of the wild-type BphA4 for NADH. In addition, this mutation project was assessed by a semiempirical prediction method of mutation effects, and the results suggested that the CRG mutant was one of the best NADPHspecific mutants.

AB - The coenzyme specificity of enzymes is one of the critical parameters for the engineered production of biological compounds using bacteria. SinceNADPH is produced abundantly in photosynthetic organisms, conversion of an NADH-specific enzyme into an NADPH-specific one is a useful approach for the efficient carbon-neutral production of biological compounds in photosynthetic organisms. In the present study, anNADH-specific ferredoxin reductase component, BphA4 of biphenyl dioxygenase BphA from Acidovorax sp. Strain KKS102, was changed to an NADPH-dependent form using a method combining structure-based systematic mutations and site-directed random mutagenesis. The resultant CRG mutant, in which Glu175-Thr176-Gln177 of an NADH-recognition loop in the wild-type BphA4 was replaced with Cys 175-Arg176-Gly177, was highly specific and active for NADPH, and its biochemical and structural properties for NADPH were nearly the same as those of the wild-type BphA4 for NADH. In addition, this mutation project was assessed by a semiempirical prediction method of mutation effects, and the results suggested that the CRG mutant was one of the best NADPHspecific mutants.

KW - Biphenyl degradation

KW - Electron transfer

KW - Flavoprotein

KW - Protein design

KW - Random mutagenesis

KW - Specificity prediction

UR - http://www.scopus.com/inward/record.url?scp=84905911662&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84905911662&partnerID=8YFLogxK

U2 - 10.1042/BJ20140384

DO - 10.1042/BJ20140384

M3 - Article

C2 - 24902961

AN - SCOPUS:84905911662

VL - 462

SP - 257

EP - 265

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -