Secretion of Pem-CMG, a peptide in the CHH/MIH/GIH family of Penaeus monodon, in Pichia pastoris is directed by secretion signal of the alpha-mating factor from Saccharomyces cerevisiae.
The CHH/MIH/GIH peptide family of black tiger prawn (Paneaus monodon) is important in shrimp reproduction and growth enhancement. In this study, the cDNA that encodes the complete peptide that is related to the CHH/MIH/GIH family (so-called, Pem-CMG) in the eyestalk of P. monodon was successfully expressed in a methylotrophic yeast Pichia pastoris under the control of an alcohol oxidase promoter. In order to obtain the secreted Pem-CMG, a secretion signal of either the Saccharomyces cerevisiae alpha-factor or Pem-CMG was employed. The results demonstrated that alphaPem-CMG, either with (alpha2EACMG) or without (alphaCMG) the Glu-Ala repeats, was secreted into the medium, while Pem-CMG with its own secretion signal failed to be secreted. The total protein amount that was secreted from the transformant that contained either alpha2EACMG or alphaMG was approximately 60 mg/l and 150 mg/l, respectively. The N-terminus of the Pem-CMG peptide of both alpha2EACMG and alphaCMG was correctly processed. This produced the mature Pem-CMG peptide.
D000818:Animals; D060829:Arthropod Proteins; D003001:Cloning, Molecular / Q000379:methods*; D005822:Genetic Vectors; D015151:Immunoblotting; D007447:Invertebrate Hormones / Q000378:metabolism; D000072235:Mating Factor; D033561:Penaeidae / Q000235:genetics* / Q000378:metabolism; D010455:Peptides / Q000235:genetics; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D060829:Arthropod Proteins; C116855:CHH-like protein, shrimp; D007447:Invertebrate Hormones; D010455:Peptides; D021382:Protein Sorting Signals; D011506:Proteins; C012445:molt-inhibiting hormone; D000072235:Mating Factor