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Purification and molecular cloning of hemocyanin from Fenneropenaeus merguiensis(de Man, 1888): response to Vibrio harveyi exposure

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Hemocyanin (HC) was purified from hemolymph of Fenneropenaeus merguiensis(de Man, 1888). Purified HC showed one band in non-denaturing PAGE with a molecular mass of 457 kDa, determined by gel filtration. Using an ELISA, HC content was 85.6% of total hemolymph protein. HC’s hexamer form contained no phenoloxidase activity under trypsin or SDS activation. FmHC, cloned from F. merguiensis, had a full-length cDNA comprised of 2128 bp with an open reading frame encoding a peptide of 661 amino acids with two Cu-binding sites. Expression of FmHCwas found only in the hepatopancreas where it was up-regulated after exposure to Vibrio harveyiinjection into shrimp and with tissue incubations. HC protein levels in the hemolymph of bacterial injected shrimp also increased. FmHCexpression was also up-regulated after shrimp were challenged with a lipopolysaccharide, which is a surface component of gram-negative bacteria. These results indicate that HC is inducible and up-regulated in response to pathogenic challenge, confirming that HC participates in the innate immune response in F. merguiensis.

Affiliations: 1: 1Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai 90112, Thailand; 2: 2Department of Biology, College of Science and Mathematics, California State University, Fresno, CA 93740-8034, U.S.A.


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