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In Silico Evaluation of Various Signal Peptides to Improve Secretion of Humulin Protein in E. coli Host

Author(s):

Soudabe Kavousipour, Mahadi Barazesh, Shiva Mohammadi*, Meghdad Abdollahpour- Alitappeh, Shirzad Fallahi, Amir Shakarami and Zeynab Hajmohammadi   Pages 1 - 13 ( 13 )

Abstract:


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.

Keywords:

Extracellular secretion, E.coli, Humulin, in silico, Signal peptide.

Affiliation:

Molecular Medicine Research Center, Hormozgan Health Institute, Hormozgan University of Medical Sciences, Bandar Abbas,, School of Paramedical Sciences Gerash Faculty of Medical Sciences, Department of biotechnology, Gerash,, Department of Medical Biotechnology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah,, Department of Biotechnology, School of Nursing, Larestan Faculty of Medical Sciences,, Hepatitis Research Center, Lorestan University of Medical Science, Khorramabad,, Department of Cardiology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad,, Student Research Committee, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran



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