Background: Escherichia coli host has been the workhorse for the production of heterologous proteins due to
simplicity of use, low cost, availability of various expression vectors, and widespread knowledge on its genetic characteristics,
but without a suitable signal sequence, this host cannot be used for production secretory proteins. Humulin is a form of
insulin used to treat hyperglycemia caused by types 1 and 2 diabetes. To improve expression and make a straightforward
production of Humulin protein, we chose a series of signal peptides.
Objective: aim our study to predict the most excellent signal peptides to express secretory Humulin in E. coli organisms.
Method: Therefore, to forecast the most excellent signal peptides for expression of Humulin in Escherichia coli, 47 signal
sequences from bacteria organisms were elected and the most imperative elements of them were studied. Hence, signal peptide
probability along with physicochemical features was evaluated by signal 4.1, and Portparam, PROSO II servers respectively.
Later, the in-silico cloning in a known pET28a plasmid system also estimated the possibility of best signal peptide+
Humulin expression in E.Coli.
Results: The outcomes demonstrated among 47 signal peptides only 2 signal peptides can be suggested as suitable signal peptides.
Conclusion: Ultimately protein yebF precursor (YEBF_ECOLI) and protein yebF precursor (YEBF_YERP3) were suggested
severally; as the most excellent signal peptides to express Humulin (With D scores 0.812 and 0.623 respectively). Although
verification of these results want experimental analysis.