High-level secretory expression, purification and characterization of Ailuropoda melanoleuca growth hormone in Pichia pastoris

Protein Expr Purif. 2008 Aug;60(2):182-7. doi: 10.1016/j.pep.2008.03.001. Epub 2008 Mar 10.

Abstract

Growth hormone is one of the most important hormones, which is involved in many reproductive processes of giant panda Ailuropoda melanoleuca. In this study, the mature peptide of A. melanoleuca growth hormone (AmGH) was successfully expressed and secreted in Pichia pastoris under the control of AOX1 promoter. The expression condition for AmGH in P. pastoris, such as the expression time, pH value and methanol concentration in the BMMY were optimized and the AmGH expression level is about 100 mg/L using GS115 recombinant under optimized condition (96 h of 1.5% methanol induction). The secreted nascent AmGH were purified using ammonium sulfate fractionation. The mature AmGH protein exhibited a molecular mass of approximately 22 kDa on SDS-PAGE. This study would provide a new opportunity for large-scale expression and purification of AmGH, which might facilitate studies on the biological activity of AmGH.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Growth Hormone / chemistry
  • Growth Hormone / genetics
  • Growth Hormone / isolation & purification*
  • Pichia / genetics
  • Polymerase Chain Reaction
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Ursidae

Substances

  • DNA Primers
  • Recombinant Proteins
  • Growth Hormone