title: Expression of biologically active crustacean hyperglycemic hormone (CHH) of Penaeus monodon in Pichia pastoris.
abstract: Crustacean hyperglycemic hormone (CHH), molt-inhibiting hormone (MIH), and gonad-inhibiting hormone (GIH) are members of a major peptide family produced from the X-organ sinus gland complex in the eyestalk of crustaceans. This peptide family plays important roles in controlling several physiologic processes such as regulation of growth and reproduction. In this study the complementary DNA encoding a peptide related to the CHH/MIH/GIH family (so-called Pem-CMG) of the black tiger prawn Penaeus monodon was successfully expressed in the yeast Pichia pastoris under the control of the AOX1 promoter. The recombinant Pem-CMG was secreted into the culture medium using the alpha-factor signal sequence; of Saccharomyces cerevisiae without the Glu-Ala-Glu-Ala spacer peptide. The amino terminus of the recombinant Pem-CMG was correctly processed as evidenced by amino-terminal peptide sequencing. The recombinant Pem-CMG was purified by reverse-phase high-performance liquid chromotography and used in a biological assay for CHH activity. The final yield of the recombinant Pem-CMG after purification was 260 micro g/L of the culture medium. Both crude and purified recombinant Pem-CMG produced from P. pastoris showed the ability to elevate the glucose level in the hemolymph of eyestalk-ablated P. monodon, which demonstrates that Pem-CMG peptide functions as hyperglycemic hormone in P. monodon.
authors.affiliation: Institute of Molecular Biology and Genetics, Mahidol University, Salaya Campus, Nakhon Pathom 73170, Thailand.
authors.forename: Supattra
authors.initials: S
authors.lastname: Treerattrakool
authors.forename: Apinunt
authors.initials: A
authors.lastname: Udomkit
authors.forename: Lily
authors.initials: L
authors.lastname: Eurwilaichitr
authors.forename: Burachai
authors.initials: B
authors.lastname: Sonthayanon
authors.forename: Sakol
authors.initials: S
authors.lastname: Panyim
mesh_terms: D000818:Animals; D060829:Arthropod Proteins; D001681:Biological Assay; D002352:Carrier Proteins / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D002874:Chromosome Mapping; D004591:Electrophoresis, Polyacrylamide Gel; D006458:Hemolymph / Q000737:chemistry; D007447:Invertebrate Hormones / Q000302:isolation & purification* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000302:isolation & purification*; D033561:Penaeidae / Q000235:genetics*; D010455:Peptides / Q000235:genetics; D010843:Pichia / Q000378:metabolism; D020539:Sequence Analysis, Protein
chemical_list: D060829:Arthropod Proteins; D002352:Carrier Proteins; D007447:Invertebrate Hormones; D009419:Nerve Tissue Proteins; D010455:Peptides; C015947:hyperglycemic hormone, crustacean; C012445:molt-inhibiting hormone; C073189:vitellogenesis inhibiting hormone