Methanocalculus chunghsingensis sp. nov., isolated from an estuary and a marine fishpond in Taiwan

Abstract

Three novel halotolerant, hydrogenotrophic methanogens, designated strains K1F9705b T , K1F9705c and O1F9704a, were isolated from an estuary in Eriln Shi, Taiwan, and from a nearby marine water aquaculture fishpond. These isolates were irregular cocci that stained Gram-negative. Strains K1F9705bT and K1F9705c were non-motile, but strain O1F9704a was weakly motile with flagella. They were able to use formate and H 2 /CO 2 to form methane, but they could not catabolize acetate, methanol, trimethylamine or secondary alcohols. Acetate was required for cell growth. Tungsten greatly stimulated the growth of strains K1F9705b T andK1F9705c, but did not affect the growth of strain O1F9704a. Optimal pH and temperature for growth of these three isolates were respectively 7?2 and 37 6C. Optimal NaCl concentration for growth was 0?5 % for strain O1F9704a and 1?0 % for strains K1F9705c and K1F9705b T .Moreover, all strains grew well at up to 8-12 % NaCl. Analysis of the 16S rRNA gene revealed that these isolates are members of the genus Methanocalculus, but are distinct from Methanocalculus taiwanensis, Methanocalculus pumilus and Methanocalculus halotolerans, with sequence similarities of 98?4, 98?3 and 98?2 %, respectively. In addition, strain K1F9705b T possessed 85, 80, 37, 29 and 10 % DNA-DNA relatedness to strain K1F9705c, strain O1F9704a, M. pumilus, M. halotolerans and M. taiwanensis, respectively. Analysis of protein profiles and the M r of surface (S)-layer glycoprotein subunits showed that these three new isolates are closely related to, but distinct from, known Methanocalculus species. A novel species, Methanocalculus chunghsingensis sp. nov., is proposed for strains K1F9705b T

Methanogenic Archaea Isolated from Taiwan's Chelungpu Fault

“…Upon inoculation into thioglycolate (TGC) medium, no growth was observed, indicating that the culture was free of anaerobic heterotrophic bacteria (30). This methanoarchaeal isolate (strain St545Mb) was deposited into DSMZ as DSM 19953, JCM as JCM 15159, and the BCRC (Taiwan) as AR10030.…”

“…Schmitz et al (47) showed that tungsten does not support synthesis of active formylmethanofuran dehydrogenase in Methanosarcina barkeri and concluded that mesophilic methanogens appear to use only molybdenum. However, mesophilic methanogens Methanofollis aquaemaris (29), Methanofollis formosanus (62), Methanocalculus taiwanensis (32), Methanocalculus chunghsingensis (30), and Methanobacterium palustre FG694aF (this study), which we isolated from estuarine, marine, and deep subsurface fault environments, all demonstrated that trace amounts of tungsten not only promoted growth but also extended the temperature, pH, and salt ranges of growth conditions. The 16S rRNA gene sequence (1,240 bp; EU293795) of strain FG694aF was determined, and phylogenetic trees were constructed using a selection of sequences from related methanogens and the environmental DNA library from GenBank (Fig.…”

Physico-chemical characteristics and methanogen communities in swine and dairy manure storage tanks: Spatio-temporal variations and impact on methanogenic activity

“…The pH value had a positive effect, probably because of (i) the reduced toxicity of anionic VFAs at neutral pH values compared to the unionized species at acidic pH (Dhaked et al, 2003) and (ii) the optimal pH for growth of several methanogens being inherently around neutrality (Boopathy and Kulpa, 1994;Lai et al, 2004).…”

Compositions of microbial communities associated with oil and water in a mesothermic oil field

“…Species of Methanomethylovorans produce methane from dimethylsulfide, methanethiol and methylamine (Lomans et al 1999), compounds that dissolve in oil rather than water, explaining the association of this genus with oil (Table 5). The hydrogenotrophic methanogens Methanocalculus and Methanoculleus require acetate as a carbon source but not as a substrate for methanogenesis (Lai et al 2002(Lai et al , 2004Mikucki et al 2003), which may explain their preferential association with the aqueous phase. The hydrogenotrophic acetogen Acetobacterium was also strongly oil-associated (Table 5).…”