title: Production of recombinant peptides as fusions with SUMO.
abstract: Recombinant production of non-native peptides requires using protein fusion technology to prevent peptide degradation by host-cell proteases. In this work, we have used SUMO protein as a fusion partner for the production of difficult-to-express, antimicrobial, self-assembling and amyloidogenic peptides using Escherichia coli. SUMO-peptide fusions were expressed as intracellular products by utilizing pET based expression vectors constructed by Life Sensors Inc., USA. Histidine tagged SUMO-peptide fusions were purified using Ni-NTA affinity chromatography. Complete (100%) cleavage of the SUMO-peptide fusion was achieved using SUMO protease-1. Our findings demonstrate that SUMO fusion technology is a promising alternative for production of peptides in E. coli. The key advantage of this technology is that the enzymatic activity of SUMO protease-1 is specific and efficient leading to inexpensive costs for cleaving the peptide fusion when compared with other fusion systems.
authors.affiliation: School of Chemical Engineering and Analytical Science, University of Manchester, Oxford Road, Manchester M139PL, United Kingdom. sata_karni@yahoo.com
authors.forename: Makkapati
authors.initials: M
authors.lastname: Satakarni
authors.forename: Robin
authors.initials: R
authors.lastname: Curtis
mesh_terms: D016229:Amyloid beta-Peptides / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism; D013058:Mass Spectrometry; D008970:Molecular Weight; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D025842:SUMO-1 Protein / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D012995:Solubility
chemical_list: D016229:Amyloid beta-Peptides; D001425:Bacterial Outer Membrane Proteins; D010446:Peptide Fragments; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D025842:SUMO-1 Protein; C075222:amyloid beta-protein (1-42); C078167:OMPA outer membrane proteins; D010447:Peptide Hydrolases