Project description:L-methionine is an important natural amino acid with broad application prospects. A novel gene encoding the enzyme with the ability to catalyze O-succinyl-L-homoserine (OSH) to L-methionine was screened and characterized. The recombinant O-succinyl-L-homoserine sulfhydrylase from Thioalkalivibrio sulfidiphilus (tsOSHS) exhibited maximum activity at 35°C and pH 6.5. OSHS displayed an excellent thermostability with a half-life of 21.72 h at 30°C. Furthermore, the activity of OSHS increased 115% after Fe2+ added. L-methionine was obtained with a total yield reaching 42.63 g/L under the concentration of O-succinyl-L-homoserine 400 mM (87.6 g/L). These results indicated that OSHS is a potential candidate for applying in the large-scale bioproduction of L-methionine.
Project description:Chemolithoautotrophic sulfur-oxidizing bacteria (SOB) are crucial key players in biotechnological processes to remove hydrogen sulfide from sour gas streams. Several different haloalkaliphilic SOB have been detected and isolated from lab- and full-scale facilities, which all performed differently considering end product yields (sulfur and sulfate) and conversion rates. Understanding and regulating bacterial community dynamics in biodesulfurization processes will enable optimization of the process operation. We developed quantitative PCR (qPCR) assays to quantify haloalkaliphilic sulfur-oxidizing gammaproteobacterial species Alkalilimnicola ehrlichii, Thioalkalivibrio sulfidiphilus, and Thioalkalibacter halophilus that dominate bacterial communities of biodesulfurization lab- and full-scale installations at haloalkaline conditions. The specificity and PCR efficiency of novel primer sets were evaluated using pure cultures of these target species. We further validated the qPCR assays by quantification of target organisms in five globally distributed full-scale biodesulfurization installations. The qPCR assays perform a sensitive and accurate quantification of Alkalilimnicola ehrlichii, Thioalkalivibrio sulfidiphilus and Thioalkalibacter halophilus, thus providing rapid and valuable insights into process performance and SOB growth dynamics in gas biodesulfurization systems.
Project description:"Thioalkalivibrio sulfidophilus" HL-EbGr7 is an obligately chemolithoautotrophic, haloalkaliphilic sulfur-oxidizing bacterium (SOB) belonging to the Gammaproteobacteria. The strain was found to predominate a full-scale bioreactor, removing sulfide from biogas. Here we report the complete genome sequence of strain HL-EbGr7 and its annotation. The genome was sequenced within the Joint Genome Institute Community Sequencing Program, because of its relevance to the sustainable removal of sulfide from bio- and industrial waste gases.
Project description:Microbial sulfur cycling and chemoautotrophy are focal points of research in cold seeps. However, limited culture-dependent and in-situ studies have described the biological features and ecological significance of chemoautotrophic sulfur-oxidizing bacteria. In this study, we isolated Guyparkeria hydrothermalis SP2, a thiosulfate/sulfide-oxidizing chemoautotrophic bacterium, from cold-seep sediment. Electron microscopy, Raman spectroscopy, and stoichiometry confirmed the efficient production of zero-valent sulfur (ZVS) by G. hydrothermalis SP2. Genomic, transcriptomic, and qRT-PCR analyses revealed its utilization of the Sox pathway for thiosulfate oxidation and the fccB gene for sulfide oxidation. Its chemoautotrophic capability mediated by the Calvin-Benson-Bassham (CBB) cycle was identified through isotopic and qRT-PCR analyses. In-situ studies demonstrated its ability to produce ZVS by oxidizing sulfide in cold seeps, with a preference for different genes compared to those under laboratory conditions. Metagenomic and metatranscriptomic analyses indicated the ubiquity of its sulfur oxidation-based chemoautotrophic pathway in cold seep sediments. Therefore, this strain holds significance for investigating sulfur oxidation-based chemoautotrophic pathways in cold seeps.