Functional expression of the globular domain of human adiponectin in Pichia pastoris

De Guo Liu, Hong Lei Liu, Tan Jing Song, Hai Yan Huang, Xi Li, Qi Qun Tang

Research output: Contribution to journalArticlepeer-review

Abstract

Adiponectin is an adipokine that predominantly synthesized and secreted from adipocytes mainly in the white adipose tissue. Here, we report that we have successfully expressed human gAdiponectin (the globular domain of adiponectin) in the methylotrophic yeast Pichia pastoris after codon optimization and established the purification procedure. The human gAdiponectin gene was designed and synthesized by PCR according to the P. pastoris preferred codons, and then inserted into the P. pastoris pPIC9K expression vector. The plasmid was electroporated into the P. pastoris strain GS115 and only the G418 resistance colonies could produce the gAdiponectin. After fermentation and purification, we could get 1.2 g of recombinant gAdiponectin (purity is ∼95%) from a 24 L culture media. The recombinant gAdiponectin is fully functional as evidenced by induction the phosphorylation of ACC in differentiated C2C12 myotubes, significantly lowering the blood glucose level and accelerating the clearance of free fatty acid in animal models.

Original languageEnglish (US)
Pages (from-to)769-775
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume363
Issue number3
DOIs
StatePublished - Nov 23 2007
Externally publishedYes

Keywords

  • Adiponectin
  • Codon optimization
  • Pichia pastoris

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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