Abstract
Exendin-4 is a naturally occurring 39 amino acid peptide that is useful for the control of Type 2 diabetes. Recombinant Exendin-4, with an extra glycine at the carboxy-terminus (Exdgly), was expressed in the methylotropic yeast Pichia pastoris. A high proportion of the Exdgly molecules secreted into medium were found to be clipped, lacking the first two amino acids (His-Gly) from the N-terminus. Disruption of the P. pastoris homolog of the Saccharomyces cerevisiae dipeptidyl aminopeptidase (STE13) gene in Pichia genome resulted in a clone that expressed N-terminally intact Exdgly. Elimination of N-terminal clipping enhanced the yield and simplified the purification of Exdgly from P. pastoris culture supernatant.
MeSH terms
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Amino Acid Sequence
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Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / chemistry*
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Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / genetics
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Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / metabolism
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Exenatide
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Molecular Sequence Data
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Peptides / chemistry*
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Peptides / genetics
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Peptides / metabolism
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Pichia
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Saccharomyces cerevisiae / enzymology
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
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Sequence Alignment
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Venoms / chemistry*
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Venoms / genetics
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Venoms / metabolism
Substances
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Peptides
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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Venoms
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Exenatide
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Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
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STE13 protein, S cerevisiae