Characterization and gene deletion analysis of four homologues of group 3 pyridine nucleotide disulfide oxidoreductases from Thermococcus kodakarensis

Harnvoravongchai, Phurt; Kobori, Hiroki; Orita, Izumi; Nakamura, Satoshi; Imanaka, Tadayuki; Fukui, Toshiaki
May 2014
Extremophiles;May2014, Vol. 18 Issue 3, p603
Academic Journal
Enzymatic characterization of the four group 3 pyridine nucleotide disulfide oxidoreductase (PNDOR) homologues TK1299, TK0304, TK0828, and TK1481 from Thermococcus kodakarensis was performed, with a focus on their CoA-dependent NAD(P)H: elemental sulfur (S) oxidoreductase (NSR) and NAD(P)H oxidase (NOX) activities. TK1299 exhibited NSR activity with a preference for NADPH and showed strict CoA-dependency similar to that of the Pyrococcus furiosus homologue PF1186. During the assays, the non-enzymatic formation of HS from S and free CoA-SH was observed, and the addition of enzyme and NADPH enhanced HS evolution. A catalytic cycle of TK1299 was proposed suggesting that CoA-SH acted to solubilize S by forming CoA persulfides, followed by reduction of an enzyme-S-S-CoA intermediate produced after both enzymatic and non-enzymatic evolution of HS from the CoA persulfide, with NADPH as an electron donor. TK1481 showed NSR activity independently of CoA-SH, implying a direct reaction with S. TK1299, TK1481, and TK0304 exhibited high NOX activity, and the NADH-dependent activities were inhibited by the addition of free CoA-SH. Multiple disruptions of the four group 3 PNDOR homologues in T. kodakarensis demonstrated that none of these homologues were essential for S-dependent growth. Many disruptants grew better than the parent strain, but a few multiple disruptants showed decreased growth properties after aerobic inoculation into a pyruvate-containing medium without S, suggesting the complicated participation of these group 3 PNDORs in sensitivity/resistance to dissolved oxygen when S was absent.


Related Articles

  • TK1656, an L-asparaginase from Thermococcus kodakarensis, a novel candidate for therapeutic applications. Chohan, Shahid Mahmood; Nisar, Muhammad Atif; Rashid, Naeem; Gharib, Ghazaleh; Bashir, Qamar; Siddiqui, Masood Ahmed // Biologia;Dec2016, Vol. 71 Issue 12, p1315 

    L-Asparaginase is a potential therapeutic agent owing to its anti-tumor activity. We have previously characterized a thermostable L-asparaginase (TK1656F from Thermococcus kodakaraensis that exhibits highest ever reported enzymatic activity, no glutaminase activity and high stability. In this...

  • Architecture and characterization of a thermostable MoxR family AAA ATPase from Thermococcus kodakarensis KOD1. Pham, Bang; Lee, Sangmin; Jia, Baolei; Kwak, Jae; Cheong, Gang-Won // Extremophiles;May2014, Vol. 18 Issue 3, p537 

    AAAATPases are ubiquitous enzymes that can function as molecular chaperones, employing the energy obtained from ATP hydrolysis to remodel macromolecules. In this report, the MoxR enzyme from Thermococcus kodakarensis KOD1 (TkMoxR) was shown to have two native forms: a two-stack hexameric ring...

  • Mechanisms of phototransformation of protochlorophyllide into chlorophyllide. Belyaeva, O.; Litvin, F. // Biochemistry (00062979);Apr2014, Vol. 79 Issue 4, p337 

    The purpose of this review is to summarize and discuss data obtained in studies on the mechanisms of the primary photophysical and photochemical reactions of protochlorophyllide photoreduction in plant materials (etiolated leaves and leaf homogenates) and in model systems. Based on the results...

  • Cytotoxin-induced NADPH oxides activation: roles in regulation of cell death. Zhang, Yongtao; Bi, Xiaolei; Jiang, Fan // Archives of Toxicology;Jul2015, Vol. 89 Issue 7, p991 

    Numerous studies have shown that a variety of cytotoxic agents can activate the NADPH oxidase system and induce redox-dependent regulation of cellular functions. Cytotoxin-induced NADPH oxidase activation may either exert cytoprotective actions (e.g., survival, proliferation, and stress...

  • NADPH oxidase p47phox siRNA attenuates adventitial fibroblasts proliferation and migration in apoE(-/-) mouse. Fang Xu; Ying Liu; Lei Shi; Wei Liu; Li Zhang; Hongjing Cai; Jie Qi; Yong Cui; Weichen Wang; Yejia Hu // Journal of Translational Medicine;2015, Vol. 13 Issue 1, p969 

    Background Reactive oxide species (ROS) derived from NADPH oxidases is involved in atherosclerosis. However, as a key component of NADPH oxidase, how p47phox regulates NADPH oxidases activity, ROS production and adventitial fibroblasts (AFs) function remains unclear. Methods p47phox in aortic...

  • BdcA, a Protein Important for Escherichia coli Biofilm Dispersal, Is a Short-Chain Dehydrogenase/Reductase that Binds Specifically to NADPH. Lord, Dana M.; Baran, Ayse Uzgoren; Wood, Thomas K.; Peti, Wolfgang; Page, Rebecca // PLoS ONE;Sep2014, Vol. 9 Issue 9, p1 

    The Escherichia coli protein BdcA (previously referred to as YjgI) plays a key role in the dispersal of cells from bacterial biofilms, and its constitutive activation provides an attractive therapeutic target for dismantling these communities. In order to investigate the function of BdcA at a...

  • Clinical, Functional and Genetic Analysis of Twenty-Four Patients with Chronic Granulomatous Disease - Identification of Eight Novel Mutations in CYBB and NCF2 Genes. Martel, Cécile; Mollin, Michelle; Beaumel, Sylvain; Brion, Jean; Coutton, Charles; Satre, Véronique; Vieville, Gaëlle; Callanan, Mary; Lefebvre, Christine; Salmon, Alexandra; Pagnier, Anne; Plantaz, Dominique; Bost-Bru, Cécile; Eitenschenck, Laurence; Durieu, Isabelle; Floret, Daniel; Galambrun, Claire; Chambost, Hervé; Michel, Gérard; Stephan, Jean-Louis // Journal of Clinical Immunology;Oct2012, Vol. 32 Issue 5, p942 

    Chronic granulomatous disease is an inherited disorder in which phagocytes lack a functional NADPH oxidase and cannot produce superoxide anions. The most common form is caused by mutations in CYBB encoding gp91 phox. We investigated 24 CGD patients and their families. Twenty-one mutations in...

  • A Hydrazine Coupled Cycling Assay Validates the Decrease in Redox Ratio under Starvation in Drosophila. Chen-Tseh Zhu; Rand, David M. // PLoS ONE;Oct2012, Vol. 7 Issue 10, Special section p1 

    A commonly used enzymatic recycling assay for pyridine nucleotides has been adapted to directly measure the NAD+/NADH redox ratio in Drosophila melanogaster. This method is also suitable for quantification of NADP+ and NADPH. The addition of a coupling reaction removing acetaldehyde produced...

  • High level expression and characterization of a thermostable lysophospholipase from Thermococcus kodakarensis KOD1. Cui, Zhicheng; Wang, Yuhan; Pham, Bang; Ping, Fangfang; Pan, Hongyu; Cheong, Gang-Won; Zhang, Shihong; Jia, Baolei // Extremophiles;Jul2012, Vol. 16 Issue 4, p619 

    Phospholipases can catalyze the hydrolysis of one or more ester and phosphodiester bonds and have a considerable interest in the food, oil leather and pharmaceutical industries. In this report, a lysophospholipase gene from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics