Glutathione S-transferase in the white shrimp Litopenaeus vannamei: Characterization and regulation under pH stress.

We first expressed a Mu-class GST from white shrimp Litopenaeus vannamei in Escherichia coli, and then characterized the purified recombinant enzyme with respect to the effects of pH, temperature on its catalytic (1-chloro-2, 4-dinitrobenzene-glutathione conjugation) activity. We also analyzed its expression profile in L. vannamei tissues, and assessed changes in Mu-GST expression, GST activity profiles and mortality rates following exposure of white shrimp to low and high pH (5.6 and 9.3, respectively). Realtime-PCR analysis showed that Mu-GST transcripts were expressed in all examined L. vannamei tissues, but were most abundant in the hepatopancreas. At low pH Mu-GST transcript levels in the hepatopancreas were highest after 12 h, and then declined to their original levels after 24 h. After 12 h they were also upregulated in haemocytes, but downregulated in the gills, and unchanged in the stomach following exposure to pH stress. Western blot analyses confirmed that the Mu-GST protein was strongly expressed in the hepatopancreas after 12 h at low pH and remain unchanged in the stomach after exposure to pH stress. pH-Related changes in GST activities in the shrimp hepatopancreas were similar to those displayed by the Mu-GST mRNA and protein profiles. In addition, the mortality of L. vannamei was higher at high pH than at low pH. These results suggest that L. vannamei Mu-GST expression is stimulated by acidic pH and that it may play important roles in detoxification of xenobiotics and antioxidant defenses.