Rhodaneses are known to catalyze in vitro the transfer of a sulfane sulfur atom from thiosulfate to cyanide with concomitant formation of thiocyanate， however， their biological functions remain speculative despite the main role is considered as detoxifying cyanide especially in animal livers. In this study， we characterized a single-domain rhodanese homologue， MnRDH1， from Macrobrachium nipponense. We found MnRDH1 with the highest expression in hemocytes. Upon Aeromonas hydrophila challenge， expression of MnRDH1 was up-regulated in various tissues， including hepatopancreas， gill， intestine and hemocytes. RNAi knockdown of MnRDH1 led to rapid increases of malondialdehyde content， which revea... More
Rhodaneses are known to catalyze in vitro the transfer of a sulfane sulfur atom from thiosulfate to cyanide with concomitant formation of thiocyanate， however， their biological functions remain speculative despite the main role is considered as detoxifying cyanide especially in animal livers. In this study， we characterized a single-domain rhodanese homologue， MnRDH1， from Macrobrachium nipponense. We found MnRDH1 with the highest expression in hemocytes. Upon Aeromonas hydrophila challenge， expression of MnRDH1 was up-regulated in various tissues， including hepatopancreas， gill， intestine and hemocytes. RNAi knockdown of MnRDH1 led to rapid increases of malondialdehyde content， which reveals that MnRDH1 deficiency causes oxidative stress. The expression of MnRDH1 in hepatopancreas was significantly increased in response to the doxorubicin-induced oxidative stress， indicating the gene is oxidative stress inducible. We transformed E. coli with MnRDH1 and the mutant MnRDH1， and found significant rhodanese activity of the recombinant protein of MnRDH1 in vitro， but detected no enzyme activity of the mutant MnRDH1. When under the oxidative insult by HO， the MnRDH1 transformed E. coli had significantly enhanced survival rates compared to those bacteria transformed with MnRDH1. In conclusion， our study demonstrates that rhodanese in M. nipponense confers oxidative stress tolerance， and thus renders an evidence for the notion that rhodanese family genes act a critical role in antioxidant defenses.
