Review
- [MINIREVIEW] Antimicrobial actions of dual oxidases and lactoperoxidase
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Demba Sarr , Eszter Tóth , Aaron Gingerich , Balázs Rada
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J. Microbiol. 2018;56(6):373-386. Published online June 1, 2018
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DOI: https://doi.org/10.1007/s12275-018-7545-1
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Abstract
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The NOX/DUOX family of NADPH oxidases are transmembrane
proteins generating reactive oxygen species as their
primary enzymatic products. NADPH oxidase (NOX) 1–5
and Dual oxidase (DUOX) 1 and 2 are members of this family.
These enzymes have several biological functions including
immune defense, hormone biosynthesis, fertilization, cell proliferation
and differentiation, extracellular matrix formation
and vascular regulation. They are found in a variety of tissues
such as the airways, salivary glands, colon, thyroid gland and
lymphoid organs. The discovery of NADPH oxidases has drastically
transformed our view of the biology of reactive oxygen
species and oxidative stress. Roles of several isoforms including
DUOX1 and DUOX2 in host innate immune defense
have been implicated and are still being uncovered. DUOX
enzymes highly expressed in the respiratory and salivary gland
epithelium have been proposed as the major sources of hydrogen
peroxide supporting mucosal oxidative antimicrobial
defenses. In this review, we shortly present data on DUOX
discovery, structure and function, and provide a detailed, upto-
date summary of discoveries regarding antibacterial, antiviral,
antifungal, and antiparasitic functions of DUOX enzymes.
We also present all the literature describing the immune
functions of lactoperoxidase, an enzyme working in
partnership with DUOX to produce antimicrobial substances.
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Journal Article
- The NADPH oxidase AoNoxA in Arthrobotrys oligospora functions as an initial factor in the infection of Caenorhabditis elegans
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Xin Li , Ying-Qian Kang , Yan-Lu Luo , Ke-Qin Zhang , Cheng-Gang Zou , Lian-Ming Liang
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J. Microbiol. 2017;55(11):885-891. Published online October 27, 2017
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DOI: https://doi.org/10.1007/s12275-017-7169-x
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Abstract
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Reactive oxygen species (ROS) produced by NADPH oxidases
can serve as signaling molecules to regulate a variety of
physiological processes in multi-cellular organisms. In the
nematophagous fungus Arthrobotrys oligospora, we found
that ROS were produced during conidial germination, hyphal
extension, and trap formation in the presence of nematodes.
Generation of an AoNoxA knockout strain demonstrated
the crucial role of NADPH oxidase in the production
of ROS in A. oligospora, with trap formation impaired in
the AoNoxA mutant, even in the presence of the nematode
host. In addition, the expression of virulence factor serine
protease P186 was up-regulated in the wild-type strain, but
not in the mutant strain, in the presence of Caenorhabditis
elegans. These results indicate that ROS derived from AoNoxA
are essential for full virulence of A. oligospora in nematodes.
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Research Support, Non-U.S. Gov'ts
- Biochemical Properties and Physiological Roles of NADP-Dependent Malic Enzyme in Escherichia coli
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Baojuan Wang , Peng Wang , Enxia Zheng , Xiangxian Chen , Hanjun Zhao , Ping Song , Ruirui Su , Xiaoning Li , Guoping Zhu
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J. Microbiol. 2011;49(5):797-802. Published online November 9, 2011
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DOI: https://doi.org/10.1007/s12275-011-0487-5
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Abstract
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Malic enzymes catalyze the reversible oxidative decarboxylation of L-malate using NAD(P)+ as a cofactor. NADP-dependent malic enzyme (MaeB) from Escherichia coli MG1655 was expressed and purified as a fusion protein. The molecular weight of MaeB was about 83 kDa, as determined by SDS-PAGE. The recombinant MaeB showed a maximum activity at pH 7.8 and 46°C. MaeB activity was dependent on the presence of Mn2+ but was strongly inhibited by Zn2+. In order to understand the physiological roles, recombinant E. coli strains (icdNADP/ΔmaeB and icdNAD/ΔmaeB) containing NADP-dependent isocitrate dehydrogenase (IDH), or engineered NAD-dependent IDH with the deletion of the maeB gene, were constructed using homologous recombination. During growth on acetate, icdNAD/ΔmaeB grew poorly, having a growth rate only 60% that of the wild-type strain (icdNADP). Furthermore, icdNADP/ΔmaeB exhibited a 2-fold greater adaptability to acetate than icdNAD/ΔmaeB, which may be explained by more NADPH production for biosynthesis in icdNADP/ΔmaeB due to its NADP-dependent IDH. These results indicated that MaeB was important for NADPH production for bacterial growth on acetate. We also observed that MaeB activity was significantly enhanced (7.83-fold) in icdNAD, which was about 3-fold higher than that in icdNADP, when switching from glucose to acetate. The marked increase of MaeB activity was probably induced by the shortage of NADPH in icdNAD. Evidently, MaeB contributed to the NADPH generation needed for bacterial growth on two carbon compounds.
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Hyun Gyu Lim, Ji Hoon Lee, Myung Hyun Noh, Gyoo Yeol Jung
Journal of Agricultural and Food Chemistry.2018; 66(16): 3998. CrossRef - Enhanced performance of the methylerythritol phosphate pathway by manipulation of redox reactions relevant to IspC, IspG, and IspH
Jia Zhou, Liyang Yang, Chonglong Wang, Eui-Sung Choi, Seon-Won Kim
Journal of Biotechnology.2017; 248: 1. CrossRef - Rational design of a synthetic Entner–Doudoroff pathway for improved and controllable NADPH regeneration
Chiam Yu Ng, Iman Farasat, Costas D. Maranas, Howard M. Salis
Metabolic Engineering.2015; 29: 86. CrossRef - Metabolic network capacity of Escherichia coli for Krebs cycle-dependent proline hydroxylation
Eleni Theodosiou, Oliver Frick, Bruno Bühler, Andreas Schmid
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- Characterization of Deinococcus radiophilus Thioredoxin Reductase Active with Both NADH and NADPH
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Hee-Jeong Seo , Young Nam Lee
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J. Microbiol. 2010;48(5):637-643. Published online November 3, 2010
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DOI: https://doi.org/10.1007/s12275-010-0283-7
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Abstract
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Thioredoxin reductase (TrxR, EC 1.6.4.5) of Deinococcus radiophilus was purified by steps of sonication, ammonium sulfate fractionation, 2'5' ADP Sepharose 4B affinity chromatography, and Sephadex G-100 gel filtration. The purified TrxR, which was active with both NADPH and NADH, gave a 368 U/mg protein of specific activity with 478-fold purification and 18% recovery from the cell-free extract. An isoelectric point of the purified enzymes was ca. 4.5. The molecular weights of the purified TrxR estimated by PAGE and gel filtration were about 63.1 and 72.2 kDa, respectively. The molecular mass of a TrxR subunit is 37 kDa. This suggests that TrxR definitely belongs to low molecular weight TrxR (L-TrxR). The Km and Vmax of TrxR for NADPH are 12.5 μM and 25 μM/min, whereas those for NADH are 30.2 μM and 192 μ M/min. The Km and Vmax for 5, 5'-dithio-bis-2-nitrobenzoic acid (DTNB, a substituted substrate for thioredoxin) are 463 μM and 756 μM/min, respectively. The presence of FAD in TrxR was confirmed with the absorbance peaks at 385 and 460 nm. The purified TrxR was quite stable from pH 3 to 9, and was thermo-stable up to 70°C. TrxR activity was drastically reduced (ca. 70%) by Cu2+, Zn2+, Hg2+, and Cd2+, but moderately reduced (ca. 50%) by Ag+. A significant inhibition of TrxR by N ethylmaleimide suggests an occurrence of cysteine at its active sites. Amino acid sequences at the N-terminus of purified TrxR are H2N-Ser-Glu-Gln-Ala-Gln-Met-Tyr-Asp-Val-Ile-Ile-Val-Gly-Gly-Gly-Pro-Ala-Gly-Leu-Thr-Ala-COOH. These sequences show high similarity with TrxRs reported in Archaea, such as Methanosarcina mazei, Archaeoglobus fulgidus etc.
- NtrC-Sensed Nitrogen Availability Is Important for Oxidative Stress Defense in Pseudomonas putida KT2440
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Sujin Yeom , Jinki Yeom , Woojun Park
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J. Microbiol. 2010;48(2):153-159. Published online May 1, 2010
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DOI: https://doi.org/10.1007/s12275-010-0075-0
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The zwf, which encodes glucose-6-phosphate dehydrogenase, is repressed by NtrC under nitrogen-limited condition. Previously, we demonstrated that induction of zwf-1 is required for protecting Pseudomonas putida cells under oxidative stress, which could be possible probably because of derepression of HexR on the zwf-1 gene under oxidative stress. These findings led us investigate that NtrC still represses the zwf-1 under nitrogen-limited oxidative stress condition, which makes cells more sensitive under such condition. Interestingly, deletion of the ntrC gene significantly reduces growth rate, but renders cells more resistant to oxidative stress, under nitrogen limited condition in P. putida. More vitality of the ntrC mutant under oxidative stress condition was also confirmed by the fluorogenic redox dye using flow cytometry. The results of transcriptome analysis demonstrated that the derepression of several oxidative stress genes along with the zwf-1 gene might confer high resistance to oxidative stress in the ntrC mutant. Here, we presented the data for the first time, showing that different sets of genes are involved in nitrogen-rich and nitrogen-limited oxidative stress conditions and NtrC-sensed nitrogen availability is one of the most important prerequisite for full cellular defense against oxidative stress in P. putida.
- Occurrence of Thioredoxin Reductase in Deinococcus Species, the UV resistant Bacteria
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Hee Jeong Seo , Young Nam Lee
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J. Microbiol. 2006;44(4):461-465.
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DOI: https://doi.org/2404 [pii]
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The occurrence of thioredoxin reductase (NAD(P)H: oxidized-thioredoxin reductase, EC 1.6.4.5, TrxR) in five mesophilic species of Deinococcus was investigated by PAGE. Each species possessed a unique TrxR pattern, for example, a single TrxR characterized D. radiopugnans while multiple forms of TrxR occurred in other Deinococcal spp. Most of TrxRs occurring in Deinococcus showed dual cofactor specificity, active with either NADH or NADPH, although the NADPH specific-TrxR was observed in D. radiophilus and D.proteolyticus.
- Regulation of fpr Gene encoding NADPH :Ferredoxin oxidoreductase by the soxRS locus in escherichia coli
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Koh, Young Sang , Chouh, Jenny , Roe, Jung Hye
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J. Microbiol. 1996;34(2):137-143.
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We isolated a promoter inducible by paraquat, a superoxide-generating agent, from Escherichia coli using a promoter-probing plasmid pRS415. From sequence analysis we found out the promoter is for fpr ENCODING nadph : ferredoxin oxidoreductase. We constructed on operon fusion of lacZ gene with fpr promoter to monitor the expression of the gene in the single-copy state. LacZ expression generators, menadione and plumbagin, also induced the expression of β-galactosidase in the fusion strain. On the other hand, no significant induction was observed by treatment with hydrogen peroxide, ethanol, and heat shock. Induction of β-galactosidase was significantly reduced by introducing a Δsox 8 :: cat of soxS3 :: Tn10 mutation into the fusion strain, indicating that fpr gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis. Possible roles of fpr induction in superoxide stress were discussed.