Purification of crustacean hyperglycemic hormone in the giant freshwater prawn Macrobrachium rosenbergii
1998
Wirawan Sitthikonkun, Srinakharinwirot Univ. Prasarnmit Campus. Bangkok (Thialnd). Faculty of Science. Dept. of Biology
Crustacean hyperglycemic hormone (CHH) of Macrobrachium rosenbergii was isolated from 5,000 frozen powdered eyestalks extracted in methanol-acetic acid. After the extract was partially purified by PrepPak C18 cartridge, it was further purified by 8 steps of RP-HPLC using four kinds of columns: C18, C8 cyano and phenyl columns; and with three solvent systems: acetonitrile (ACN)/trifluoroacetic acid, ACNheptafluorobutyric acid and ACNtriethylamine. Detection of CHH during purification was done by injection of the extracts into eyeablated prawns and determination the ability of the extract on elevation of glucose in hemolymph. The complete amino acid sequence analysis was performed on one isoform of CHH consisting of 71 residues. The sequence of the CHH shows considerable similarity (60 percent) to that of CHH of the crayfishes Orconectes limosus and Procambarus bouvieri, the american lobster Homarus americanus and the shore crab Carcinus maenas and less similarity (45 percent) to that of CHH of Penaeus japonicus. It is expected that there might be more than one isoform of CHH in M. rosenbergii.
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