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Functional expression of the globular domain of human adiponectin in Pichia pastoris
- Source :
- Biochemical and Biophysical Research Communications. 363:769-775
- Publication Year :
- 2007
- Publisher :
- Elsevier BV, 2007.
-
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 1.2g of recombinant gAdiponectin (purity is approximately 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.
- Subjects :
- Blood Glucose
Genetic Vectors
Immunoblotting
Molecular Sequence Data
Biophysics
White adipose tissue
Biochemistry
Pichia
Cell Line
Diabetes Mellitus, Experimental
Pichia pastoris
law.invention
Myoblasts
Mice
Transformation, Genetic
Plasmid
law
Sequence Homology, Nucleic Acid
Animals
Humans
Amino Acid Sequence
Phosphorylation
Codon
Molecular Biology
chemistry.chemical_classification
Binding Sites
Expression vector
Base Sequence
Dose-Response Relationship, Drug
Adiponectin
biology
Fatty Acids
Fatty acid
Cell Biology
biology.organism_classification
Molecular biology
Recombinant Proteins
Yeast
Diabetes Mellitus, Type 1
chemistry
Recombinant DNA
Acetyl-CoA Carboxylase
Subjects
Details
- ISSN :
- 0006291X
- Volume :
- 363
- Database :
- OpenAIRE
- Journal :
- Biochemical and Biophysical Research Communications
- Accession number :
- edsair.doi.dedup.....05e53bd62d7bcb44e417afadf3b11d6b