scholarly journals Electron carriers involved in autotrophic and heterotrophic acetogenesis in the thermophilic bacterium Thermoanaerobacter kivui

Extremophiles ◽  
2021 ◽  
Author(s):  
Alexander Katsyv ◽  
Surbhi Jain ◽  
Mirko Basen ◽  
Volker Müller
Keyword(s):  
Affinity Chromatography ◽  
Electron Donor ◽  
Electron Flow ◽  
Model Organism ◽  
Thermophilic Bacterium ◽  
Organic Substrates ◽  
Cell Free Extract ◽  
Electron Carriers ◽  
Thermoanaerobacter Kivui

AbstractThermoanaerobacter kivui is an acetogenic model organism that reduces CO2 with electrons derived from H2 or CO, or from organic substrates in the Wood–Ljugdahl pathway (WLP). For the calculation of ATP yields, it is necessary to know the electron carriers involved in coupling of the oxidative and reductive parts of metabolism. Analyses of key catabolic oxidoreductases in cell-free extract (CFE) or with purified enzymes revealed the physiological electron carriers involved. The glyceraldehyde-3-phosphate dehydrogenase (GA3P-DH) assayed in CFE was NAD+-specific, NADP+ was used with less than 4% and ferredoxin (Fd) was not used. The methylene-THF dehydrogenase was NADP+-specific, NAD+ or Fd were not used. A Nfn-type transhydrogenase that catalyzes reduced Fd-dependent reduction of NADP+ with NADH as electron donor was also identified in CFE. The electron carriers used by the potential electron-bifurcating hydrogenase (HydABC) could not be unambiguously determined in CFE for technical reasons. Therefore, the enzyme was produced homologously in T. kivui and purified by affinity chromatography. HydABC contained 33.9 ± 4.5 mol Fe/mol of protein and FMN; it reduced NADP+ but not NAD+. The methylene-THF reductase (MetFV) was also produced homologously in T. kivui and purified by affinity chromatography. MetFV contained 7.2 ± 0.4 mol Fe/mol of protein and FMN; the complex did neither use NADPH nor NADH as reductant but only reduced Fd. In sum, these analysis allowed us to propose a scheme for entire electron flow and bioenergetics in T. kivui.

The chemistry of stabilized clusters

1982 ◽  
Vol 308 (1501) ◽  
pp. 27-34 ◽  
Keyword(s):  
Electron Donor ◽  
Organic Molecules ◽  
Metal Cluster ◽  
Organic Substrates ◽  
Insertion Reactions ◽  
Donor Ligand ◽  
Small Organic Molecules ◽  
Wide Range ◽  
Metal Metal ◽  
High Yields

Studies of the chemistry of metal cluster complexes and, in particular, their reactions with small organic molecules, have been confined to relatively few systems. Among the reasons for this are: (i) not many clusters are easily synthesized in high yields; (ii) their reactions often give a multitude of products that are difficult to separate and characterize; (iii) the conditions required to bring about reactions often lead to fragmentation of the cluster into lower nuclearity (often mononuclear) species. One cluster whose chemistry has been extensively studied is [Os 3 H 2 (CO) 10 ]. This can be synthesized in high yields from [Os 3 (CO) 12 ] + H 2 (Knox et al. 1975) and reacts readily under mild conditions with a wide range of electron-donor molecules by virtue of its coordinative unsaturation (Shapley et al. 1975; Deeming & Hasso 1976; Adams & Golembeski 1979). Formally, one may consider that a metal—metal double bond is present, which is reduced to a single bond on coordination of an additional two-electron donor ligand such as an organophosphine. The presence of metal—hydrogen bonds in this cluster and the cluster’s ability to coordinate organic substrates enable it to undergo a wide variety of insertion reactions, leading to products that may be regarded as intermediates in the reduction of organic molecules by clusters (Deeming & Hasso 1975; Keister & Shapley 1975).

scholarly journals Lactose-Inducible System for Metabolic Engineering of Clostridium ljungdahlii

2014 ◽  
Vol 80 (8) ◽  
pp. 2410-2416 ◽  
Author(s):  
Areen Banerjee ◽  
Ching Leang ◽  
Toshiyuki Ueki ◽  
Kelly P. Nevin ◽  
Derek R. Lovley
Keyword(s):  
Ethanol Production ◽  
Genetic Manipulation ◽  
Electron Flow ◽  
Model Organism ◽  
Inducible Expression ◽  
Carbon Flow ◽  
Wild Type ◽  
Content Type ◽  
Microbial Electrosynthesis ◽  
Clostridium Ljungdahlii

ABSTRACTThe development of tools for genetic manipulation ofClostridium ljungdahliihas increased its attractiveness as a chassis for autotrophic production of organic commodities and biofuels from syngas and microbial electrosynthesis and established it as a model organism for the study of the basic physiology of acetogenesis. In an attempt to expand the genetic toolbox forC. ljungdahlii, the possibility of adapting a lactose-inducible system for gene expression, previously reported forClostridium perfringens, was investigated. The plasmid pAH2, originally developed forC. perfringenswith agusAreporter gene, functioned as an effective lactose-inducible system inC. ljungdahlii. Lactose induction ofC. ljungdahliicontaining pB1, in which the gene for the aldehyde/alcohol dehydrogenase AdhE1 was downstream of the lactose-inducible promoter, increased expression ofadhE130-fold over the wild-type level, increasing ethanol production 1.5-fold, with a corresponding decrease in acetate production. Lactose-inducible expression ofadhE1in a strain in whichadhE1and theadhE1homologadhE2had been deleted from the chromosome restored ethanol production to levels comparable to those in the wild-type strain. Inducing expression ofadhE2similarly failed to restore ethanol production, suggesting thatadhE1is the homolog responsible for ethanol production. Lactose-inducible expression of the four heterologous genes necessary to convert acetyl coenzyme A (acetyl-CoA) to acetone diverted ca. 60% of carbon flow to acetone production during growth on fructose, and 25% of carbon flow went to acetone when carbon monoxide was the electron donor. These studies demonstrate that the lactose-inducible system described here will be useful for redirecting carbon and electron flow for the biosynthesis of products more valuable than acetate. Furthermore, this tool should aid in optimizing microbial electrosynthesis and for basic studies on the physiology of acetogenesis.

scholarly journals Synchronous recovery of iron and electricity using a single chamber air-cathode microbial fuel cell

RSC Advances ◽  
2017 ◽  
Vol 7 (21) ◽  
pp. 12503-12510 ◽  
Author(s):  
Xiufen Li ◽  
Yan Zheng ◽  
Pengfei Nie ◽  
Yueping Ren ◽  
Xinhua Wang ◽  
...  
Keyword(s):  
Fuel Cell ◽  
Microbial Fuel Cell ◽  
Electron Donor ◽  
Electricity Generation ◽  
Metal Recovery ◽  
Organic Substrates ◽  
Air Cathode ◽  
Single Chamber ◽  
Attractive Option

In recent years, microbial fuel cell (MFC) technology has become an attractive option for metal recovery/removal at the cathode combined with electricity generation, using organic substrates as electron donor at the anode.

Effect of Magnesium and Calcium Ions on the Photoelectron Transport Activity of Low-Salt Suspended Hydrilla verticillata Thylakoids: Possible Sites of Cation Interaction

1998 ◽  
Vol 53 (9-10) ◽  
pp. 849-856
Author(s):  
Sujata R. Mishra ◽  
Surendra Chandra Sabat
Keyword(s):  
Electron Transport ◽  
Photosystem Ii ◽  
Electron Donor ◽  
Water Oxidation ◽  
Electron Flow ◽  
Hydrilla Verticillata ◽  
Transport Activity ◽  
Low Salt ◽  
Electron Transport Activity ◽  
Stimulation Of

Stimulatory effect of divalent cations like calcium (Ca2+) and magnesium (Mg2+) was investigated on electron transport activity of divalent cation deficient low-salt suspended (LS) thylakoid preparation from a submerged aquatic angiosperm, Hydrilla verticillata. Both the cations stimulated electron transport activity of LS-suspended thylakoids having an intact water oxidation complex. But in hydroxylamine (NH2OH) - or alkaline Tris - washed thylakoid preparations (with the water oxidation enzyme impaired), only Ca2+ dependent stimulation of electron transport activity was found. The apparent Km of Ca2+ dependent stimulation of electron flow from H2O (endogenous) or from artificial electron donor (exogenous) to dichlorophenol indophenol (acceptor) was found to be identical. Calcium supported stimulation of electron transport activity in NH2OH - or Tris - washed thylakoids was electron donor selective, i.e., Ca2+ ion was only effective in electron flow with diphenylcarbazide but not with NH2OH as electron donor to photosystem II. A magnesium effect was observed in thylakoids having an intact water oxidation complex and the ion became unacceptable in NH2OH - or Tris - washed thylakoids. Indirect experimental evidences have been presented to suggest that Mg2+ interacts with the water oxidation complex, while the Ca2+ interaction is localized betw een Yz and reaction center of photosystem II.

A method for on-line measurement of wastewater organic substrate oxidation level during aerobic heterotrophic respiration

2013 ◽  
Vol 67 (8) ◽  
pp. 1809-1815
Author(s):  
E. A. Rudelle ◽  
J. Vollertsen ◽  
T. Hvitved-Jacobsen ◽  
A. H. Nielsen
Keyword(s):  
Electron Flow ◽  
Domestic Wastewater ◽  
Substrate Oxidation ◽  
Heterotrophic Respiration ◽  
Fiber Optic Sensor ◽  
Organic Substrates ◽  
Dissolved Carbon ◽  
Oxidation Level ◽  
Line Measurement ◽  
On Line

A method for on-line measurement of the organic carbon oxidation level (OXC) during aerobic heterotrophic respiration in domestic wastewater was developed and tested. The method is based on batch incubation of sewer wastewater in an intermittently aerated respirometric reactor. Between aeration cycles, measured pH, dissolved oxygen (DO) and dissolved carbon dioxide (CO2) were used to calculate electron flow accepted by DO and the resulting production of dissolved inorganic carbon (DIC). The CO2 production was measured using a novel fiber-optic sensor based on luminescence quenching. The method was tested on domestic wastewater with a relatively high pH and alkalinity. From the DO and DIC measurements, it was possible to evaluate substrate oxidation levels with a temporal resolution of less than an hour. Addition of organic substrates during the experiments confirmed the method's applicability. The substrates tested included ethanol (OXC = −2), glucose (OXC = 0) and oxalic acid (OXC = 3).

Heat-stress stimulation of electron flow in a photosystem I submembrane fraction

1990 ◽  
Vol 68 (7-8) ◽  
pp. 999-1004 ◽  
Author(s):  
Natalie Boucher ◽  
Johanne Harnois ◽  
Robert Carpentier
Keyword(s):  
Heat Stress ◽  
Cytochrome B ◽  
Electron Donor ◽  
Photosystem I ◽  
Conformational Changes ◽  
Electron Flow ◽  
Primary Electron ◽  
Primary Electron Donor ◽  
Terminal Electron Acceptor ◽  
Stimulation Of

Oxygen uptake using methyl viologen as the terminal electron acceptor was recorded in digitonin-derived photosystem I submembrane fractions incubated at either 25 or 50 °C. A two- to four-fold heat-stress stimulation of electron flow was detected at 50 °C when reduced 2,6-dichlorophenol-indophenol was used as the primary electron donor. However, no stimulation was seen with N,N,N′,N′ -tetramethylphenylenediamine as the donor. The stimulation was enhanced by specific cations (Mg2+, Na+, K+), but not by Mn2 or Ca2+. The enhancement obtained with Mg2+ could be eliminated by incubating for a prolonged period. It is proposed that the observed heat-stress stimulation is due to a conformational change at the level of the cytochrome b6–f complex. This change increased the affinity of the protein complex for 2,6-dichlorophenol-indophenol at its oxidation sites. The involvement of a conformational modification is demonstrated by the absence of heat-stress stimulation in submembrane fractions immobilized in an albumin–glutaraldehyde cross-linked matrix.Key words: electron transport, photosystem I, heat stress, cytochrome b6–f complex, conformational changes, electron donor.

scholarly journals Uranium(VI) Reduction by Anaeromyxobacter dehalogenans Strain 2CP-C

2006 ◽  
Vol 72 (5) ◽  
pp. 3608-3614 ◽  
Author(s):  
Qingzhong Wu ◽  
Robert A. Sanford ◽  
Frank E. L�ffler
Keyword(s):  
Electron Donor ◽  
Ferric Iron ◽  
Model Organism ◽  
Complex Interaction ◽  
Cell Suspensions ◽  
Electron Acceptors ◽  
C Cells ◽  
Inhibitory Effects ◽  
Halogenated Compounds ◽  
Metabolic Versatility

ABSTRACT Previous studies demonstrated growth of Anaeromyxobacter dehalogenans strain 2CP-C with acetate or hydrogen as the electron donor and Fe(III), nitrate, nitrite, fumarate, oxygen, or ortho-substituted halophenols as electron acceptors. In this study, we explored and characterized U(VI) reduction by strain 2CP-C. Cell suspensions of fumarate-grown 2CP-C cells reduced U(VI) to U(IV). More-detailed growth studies demonstrated that hydrogen was the required electron donor for U(VI) reduction and could not be replaced by acetate. The addition of nitrate to U(VI)-reducing cultures resulted in a transitory increase in U(VI) concentration, apparently caused by the reoxidation of reduced U(IV), but U(VI) reduction resumed following the consumption of N-oxyanions. Inhibition of U(VI) reduction occurred in cultures amended with Fe(III) citrate, or citrate. In the presence of amorphous Fe(III) oxide, U(VI) reduction proceeded to completion but the U(VI) reduction rates decreased threefold compared to control cultures. Fumarate and 2-chlorophenol had no inhibitory effects on U(VI) reduction, and both electron acceptors were consumed concomitantly with U(VI). Since cocontaminants (e.g., nitrate, halogenated compounds) and bioavailable ferric iron are often encountered at uranium-impacted sites, the metabolic versatility makes Anaeromyxobacter dehalogenans a promising model organism for studying the complex interaction of multiple electron acceptors in U(VI) reduction and immobilization.

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