Authors : Pyne P, Alam M, Rameez MJ, Mandal S, Sar A, Mondal N, Debnath U, Mathew B, Misra AK, Mandal AK, Ghosh W
Publication Year : 2018
The SoxXAYZB(CD)2 -mediated pathway of bacterial sulfur-chemolithotrophy explains the oxidation of thiosulfate, sulfide, sulfur and sulfite, but not tetrathionate. Advenella kashmirensis, which oxidizes tetrathionate to sulfate, besides forming it as an intermediate during thiosulfate-oxidation, possesses a soxCDYZAXOB operon. Knock-out-mutations proved that only SoxBCD is involved in A. kashmirensis tetrathionate-oxidation, whereas thiosulfate-to-tetrathionate-conversion is Sox-independent. Expression of two glutathione-metabolism-related proteins increased under chemolithotrophic conditions, as compared to the chemoorganotrophic one. Substrate-dependent oxygen-consumption pattern of whole-cells, and sulfur-oxidizing enzyme activities of cell-free-extracts, measured in the presence/absence of thiol-inhibitors/glutathione, corroborated glutathione-involvement in tetrathionate-oxidation. Furthermore, proteome analyses detected a sulfite:acceptor oxidoreductase (SorAB) exclusively under chemolithotrophic conditions, while expression of a methanol dehydrogenase (XoxF) homolog, subsequently named thiol dehydrotransferase (ThdT), was found to increase three- and ten-fold during thiosulfate-to-tetrathionate-conversion and tetrathionate-oxidation, respectively. A thdT-knocked-out mutant did not oxidize tetrathionate, but converted half of the supplied 40-mM-S thiosulfate to tetrathionate. Knock-out of another thiosulfate dehydrogenase (tsdA) gene proved that both ThdT and TsdA individually converted ~20-mM-S thiosulfate to tetrathionate. The overexpressed and isolated ThdT protein exhibited PQQ-dependent thiosulfate dehydrogenation, whereas its PQQ-independent thiol-transfer activity involving tetrathionate and glutathione potentially produced a glutathione:sulfodisulfane adduct and sulfite. SoxBCD and SorAB were hypothesized to oxidize the aforesaid adduct and sulfite, respectively.