Purpose:The human somatropin is a single-chain polypeptide with a pivotal role in various biological processes. AlthoughEscherichia coliis considered as a preferred host for the production of human somatropin, the high expression of this protein inE. coliresults in the accumulation of protein as inclusion bodies. Periplasmic expression using signal peptides could be used to overcome the formation of inclusion bodies; still, the efficiency of each of the signal peptides in periplasmic transportation is varied and often is protein specific. The present study aimed to usein silicoanalysis to identify an appropriate signal peptide for the periplasmic expression of human somatropin inE. coli.Methods:A library containing 90 prokaryotic and eukaryotic signal peptides were collected from the signal peptide database, and each signal’s characteristics and efficiency in connection with the target protein were analyzed by different software. The prediction of the secretory pathway and the cleavage position was determined by the signalP5 server. Physicochemical properties, including molecular weight, instability index, gravity, and aliphatic index, were investigated by ProtParam software.Results:The results of the present study showed that among all the signal peptides studied, five signal peptides ynfB, sfaS, lolA, glnH, and malE displayed high scores for periplasmic expression of human somatropin inE. coli, respectively.Conclusion:In conclusion, the results indicated that in-silico analysis could be used for the identification of suitable signal peptides for the periplasmic expression of proteins. Further laboratory studies can evaluate the accuracy of the results ofin silicoanalysis.