Recombinant insulin hormone production in Pichia pastoris

Insulin is a peptide hormone, produced by beta cells of the pancreas and regulates the glucose homeostasis in the body Diabetes mellitus is a metabolic disease in which a person has high blood sugar, either because the pancreas does not produce enough insulin, or because cells do not respond to produced insulin. At the present time, insulin is used to treatment of this disease. İnsulin hormone is first protein produced recombinantly by using genetic engineering techniques. Firstly, it was produced in E. coli and Saccharomyces cerevisia, but recently the number of studies that used Pichia pastoris in the production of recombinant insulin increase. This study was aimed that the production of human insulin hormone precursor (IP) under the inducible AOX1 promotor of metilotrophic yeast P. pastoris. The small scale protein production studies have been done with the clones in shake flasks for 120 hours. During the shake flask production study, the samples was taken at specific time intervals and obtained supernatants for SDS-PAGE and Western blot analysis to confirm protein production. The results of SDS-PAGE and Western blot analysis showed that the protein was produced in shake flask but was not secreted out of the cell. When the protein production was done in fermentor, the analysis showed that produced protein was secreted out of the cell.