The genus Singulisphaera accommodates stalk-free planctomycetes with spherical cells, which multiply by budding, possess complex cell organization, large genomes, and colonize a wide spectrum of terrestrial environments. Two described species of this genus, S. acidiphila and S. rosea, were isolated from acidic peatlands and characterized as mildly acidophilic bacteria. Here, we characterize two neutrophilic Singulisphaera isolates from a boreal fen, strains Ch08T and PoTT. We also obtained and analyzed the previously unavailable genome sequence of S. rosea S26T. As confirmed by genome analyses and substrate utilization tests, all strains of Singulisphaera species were capable of growth on amorphous chitin due to the possession of chitinases affiliated with the glycoside hydrolase family GH18. The gene encoding chitinase from strain PoTT was expressed in Escherichia coli, and the endochitinase activity of the recombinant enzyme was confirmed. The genomes also contained gene clusters encoding Planctomycetal-type bacterial microcompartment (BMC) organelles, suggesting participation of Singulisphaera species in degradation of plant-derived polysaccharides. Transcriptome analysis performed with glucose- and L-fucose-grown cells of strain PoTT showed up-regulation of these BMC-encoding genes during growth on L-fucose. Tyrosinases involved in the oxidative removal of phenolic compounds were present only in S. acidiphila MOB10T. All Singulisphaera genomes contained the genes of the recently identified pathway of ornithine lipid biosynthesis. Based on the reported results, we describe two novel species, S. photorubra and S. chitinilytica with the type strains Ch08T (=KCTC 102485T = LMG 34153T) and PoTT (=KCTC 102290T = VKM B-3855T), respectively. The emended description of the genus Singulisphaera is also proposed.
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