Isolation of a cytochrome-deficient mutant strain of Sporomusa sphaeroides not capable of oxidizing methyl groups

Kamlage, Beate; Blaut, Michaël

doi:10.1128/jb.175.10.3043-3050.1993

Cited by 21 publications

(9 citation statements)

References 40 publications

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“…The absence of cytochrome b may explain why strain AMP cannot grow acetogenically on H 2 / CO 2 or formate. A cytochrome b-deWcient mutant of Sporomusa sphaeroides BK824 (Kamlage and Blaut 1993) and M. thermoacetica (Fröstl et al 1996) that fails to express cytochrome b in the presence of nitrate, were also not able to grow on these substrates in the basal media. In Acetobacterium woodii and Thermoanaerobacter kivui energy is conserved in a sodium ion gradient, in a pathway that does not involve cytochrome b (Müller and Gottschalk 1994).…”

Section: Discussionmentioning

confidence: 99%

Atypical one-carbon metabolism of an acetogenic and hydrogenogenic Moorella thermoacetica strain

et al. 2008

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A thermophilic spore-forming bacterium (strain AMP) was isolated from a thermophilic methanogenic bioreactor that was fed with cobalt-deprived synthetic medium containing methanol as substrate. 16S rRNA gene analysis revealed that strain AMP was closely related to the acetogenic bacterium Moorella thermoacetica DSM 521(T) (98.3% sequence similarity). DNA-DNA hybridization showed 75.2 +/- 4.7% similarity to M. thermoacetica DSM 521(T), suggesting that strain AMP is a M. thermoacetica strain. Strain AMP has a unique one-carbon metabolism compared to other Moorella species. In media without cobalt growth of strain AMP on methanol was only sustained in coculture with a hydrogen-consuming methanogen, while in media with cobalt it grew acetogenically in the absence of the methanogen. Addition of thiosulfate led to sulfide formation and less acetate formation. Growth of strain AMP with CO resulted in the formation of hydrogen as the main product, while other CO-utilizing Moorella strains produce acetate as product. Formate supported growth only in the presence of thiosulfate or in coculture with the methanogen. Strain AMP did not grow with H(2)/CO(2), unlike M. thermoacetica (DSM 521(T)). The lack of growth with H(2)/CO(2) likely is due to the absence of cytochrome b in strain AMP.

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Section: Discussionmentioning

confidence: 99%

Atypical one-carbon metabolism of an acetogenic and hydrogenogenic Moorella thermoacetica strain

et al. 2008

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“…In contrast with previous results [17], we detected glycine reductase activity even during growth on serine. The formation of glycine from sarcosine or betaine by transmethylation as observed in Sporomusa species [31] seems rather unlikely, as no corresponding enzyme activity was found (unpublished results). Generation of glycine via serine hydroxymethyltransferase is conceivable, but is unlikely because of the low activity of this enzyme [1].…”

Section: Discussionmentioning

confidence: 99%

Substrate‐specific selenoprotein B of glycine reductase from  Eubacterium acidaminophilum

Wagner

Sonntag

Grimm³

et al. 1999

European Journal of Biochemistry

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The substrate-specific selenoprotein B of glycine reductase (PB glycine ) from Eubacterium acidaminophilum was purified and characterized. The enzyme consisted of three different subunits with molecular masses of about 22 (a), 25 (b) and 47 kDa (g), probably in an a 2 b 2 g 2 composition. PB glycine purified from cells grown in the presence of [ 75 Se]selenite was labeled in the 47-kDa subunit. The 22-kDa and 47-kDa subunits both reacted with fluorescein thiosemicarbazide, indicating the presence of a carbonyl compound. This carbonyl residue prevented N-terminal sequencing of the 22-kDa (a) subunit, but it could be removed for Edman degradation by incubation with o-phenylenediamine.A DNA fragment was isolated and sequenced which encoded b and a subunits of PB glycine (grdE), followed by a gene encoding selenoprotein A (grdA2) and the g subunit of PB glycine (grdB2). The cloned DNA fragment represented a second GrdB-encoding gene slightly different from a previously identified partial grdB1-containing fragment. Both grdB genes contained an in-frame UGA codon which confirmed the observed selenium content of the 47-kDa (g) subunit. Peptide sequence analyses suggest that grdE encodes a proprotein which is cleaved into the previously sequenced N-terminal 25-kDa (b) subunit and a 22-kDa (a) subunit of PB glycine . Cleavage most probably occurred at an -Asn-Cys-site concomitantly with the generation of the blocking carbonyl moiety from cysteine at the a subunit.

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“…It has been suggested that b-type cytochromes, which are present in many Sporomusa species (including strain TmAO3 T ), are involved in fumarate reduction (Diekert, 1992;Diekert & Wohlfarth, 1994). However, b-type cytochromes are also present in species that do not grow on dicarboxylic acids (S. termitida, Sporomusa ovata and S. sphaeroides) and there is evidence for S. sphaeroides that b-type cytochromes are also involved in the oxidation of methyl groups (Kamlage & Blaut, 1993).…”

Section: Physiology and Ecologymentioning

confidence: 99%

Sporomusa aerivorans sp. nov., an oxygen-reducing homoacetogenic bacterium from the gut of a soil-feeding termite

Boga

Ludwig

Brune

2003

International Journal of Systematic and Evolutionary Microbiology

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Previously undescribed, homoacetogenic bacteria were isolated from gut homogenates of the soil-feeding termite Thoracotermes macrothorax. The isolates were slightly curved, banana-shaped rods (0?6-0?761?3-7?0 mm) and were motile by one or more lateral flagella. In older cultures, cells formed club-like sporangia that developed into terminal, heat-resistant endospores. Cells stained Gram-positive but were Gram-negative in the KOH test. The isolates were mesophilic and grew homoacetogenically on H 2 /CO 2 and L-lactate. Strain TmAO3 T , which was characterized further, also grew homoacetogenically on pyruvate, citrate, L-alanine, D-mannitol, ethanol, formate and methanol. Succinate was decarboxylated to propionate; fumarate, L-malate and oxaloacetate were fermented to propionate and acetate. Hexoses were not used as substrates.Resting cells had a large capacity for hydrogen-dependent oxygen reduction [826 nmol min "1(mg protein) "1 ], which enabled them to initiate growth in non-reduced basal medium that originally contained up to 1?5 kPa oxygen in the headspace, although growth commenced only after the medium had been rendered anoxic. Redox difference spectra of cell extracts indicated the presence of membrane-bound b-type cytochrome(s). Comparative 16S rRNA gene sequence analysis revealed that strain TmAO3 T belongs to a subgroup of the phylum of Gram-positive bacteria that is characterized by low DNA G+C content and a Gram-negative cell wall. It is related most closely to representatives of the genus Sporomusa. Based on morphological and physiological properties and on 16S rRNA gene sequence similarity of 94-97 % to other Sporomusa species, the isolates are assigned to Sporomusa aerivorans sp. nov. (type strain, TmAO3Reductive acetogenesis from H 2 and CO 2 in gut homogenates of wood-feeding termites was first reported by Breznak & Switzer (1986). In the following years, the presence of homoacetogenic activity has been demonstrated for a large number of termite species from all major feeding guilds, including representatives of wood-feeding, funguscultivating and soil-feeding termites (Brauman et al., 1992). Although reductive acetogenesis was always out-competed as a hydrogen sink by methanogenesis in gut homogenates of soil-feeding termites (Brauman et al., 1992), microinjection of H 14 CO 3 2 into intact hindguts of soil-feedingCubitermes spp. has identified a high potential for reductive acetogenesis (Tholen & Brune, 1999), indicating that the contribution of reductive acetogenesis to overall electron flow in the guts of soil-feeding termites may be larger than expected.In order to define the metabolic potential of termite gut homoacetogens and to identify specific adaptations to the gut environment, it is necessary to study these bacteria in pure culture . However, only five strains of homoacetogenic bacterium have so far been isolated from termite guts, four of them from wood-feeding species. They comprise Sporomusa termitida and Acetonema longum from Nasutitermes nigriceps and Pterotermes occidentis (Bre...

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Isolation of a cytochrome-deficient mutant strain of Sporomusa sphaeroides not capable of oxidizing methyl groups

Cited by 21 publications

References 40 publications

Atypical one-carbon metabolism of an acetogenic and hydrogenogenic Moorella thermoacetica strain

Atypical one-carbon metabolism of an acetogenic and hydrogenogenic Moorella thermoacetica strain

Substrate‐specific selenoprotein B of glycine reductase from  Eubacterium acidaminophilum

Sporomusa aerivorans sp. nov., an oxygen-reducing homoacetogenic bacterium from the gut of a soil-feeding termite

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Isolation of a cytochrome-deficient mutant strain of Sporomusa sphaeroides not capable of oxidizing methyl groups

Cited by 21 publications

References 40 publications

Atypical one-carbon metabolism of an acetogenic and hydrogenogenic Moorella thermoacetica strain

Atypical one-carbon metabolism of an acetogenic and hydrogenogenic Moorella thermoacetica strain

Substrate‐specific selenoprotein B of glycine reductase from Eubacterium acidaminophilum

Sporomusa aerivorans sp. nov., an oxygen-reducing homoacetogenic bacterium from the gut of a soil-feeding termite

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Substrate‐specific selenoprotein B of glycine reductase from  Eubacterium acidaminophilum