A new prokaryotic expression vector for the expression of antimicrobial peptide abaecin using SUMO fusion tag

Abstract

Background Despite the growing demand for antimicrobial peptides (AMPs) for clinical use as an alternative approach against antibiotic-resistant bacteria, the manufacture of AMPs relies on expensive, small-scale chemical methods. The small ubiquitin-related modifier (SUMO) tag is industrially practical for increasing the yield of recombinant proteins by increasing solubility and preventing degradation in expression systems. Results A new vector system, pKSEC1, was designed to produce AMPs, which can work in prokaryotic systems such as Escherichia coli and plant chloroplasts. 6xHis was tagged to SUMO for purification of SUMO-fused AMPs. Abaecin, a 34-aa-long antimicrobial peptide from honeybees, was expressed in a fusion form to 6xHis-SUMO in a new vector system to evaluate the prokaryotic expression platform of the antimicrobial peptides. The fusion sequences were codon-optimized in three different combinations and expressed in E. coli . The combination of the native SUMO sequence with codon-optimized abaecin showed the highest expression level among the three combinations, and most of the expressed fusion proteins were detected in soluble fractions. Cleavage of the SUMO tag by sumoase produced a 29-aa-long abaecin derivative with a C-terminal deletion. However, this abaecin derivative still retained the binding sequence for its target protein, DnaK. Antibacterial activity of the 29-aa long abaecin was tested against Bacillus subtilis alone or in combination with cecropin B. The combined treatment of the abaecin derivative and cecropin B showed bacteriolytic activity 2 to 3 times greater than that of abaecin alone. Conclusions Using a SUMO-tag with an appropriate codon-optimization strategy could be an approach for the production of antimicrobial peptides in E.coli without affecting the viability of the host cell. Electronic supplementary material The online version of this article (10.1186/s12896-019-0506-x) contains supplementary material, which is available to authorized users.

A Comprehensive Antimicrobial Activity Evaluation of the Recombinant Microcin J25 Against the Foodborne Pathogens Salmonella and E. coli O157:H7 by Using a Matrix of Conditions

“…Therefore, the development of alternative antimicrobial agents for human health, food safety, and animal health is of critical importance. Nowadays, recombinant AMPs is promising alternatives to conventional antibiotics for the treatment of pathogens infections because of chemical synthesis is not suited for synthesis of large AMPs and is not scalable for large-scale production for clinical studies and commercial use (Cao et al, 2018; Wei et al, 2018; Kim et al, 2019; Meng et al, 2019).…”

“…But, in recent years, chemical synthesis of AMPs is extreme costly and is not scalable for large-scale AMP production. Recombinant expression was always used to produce AMPs in many laboratories (Cao et al, 2018; Wei et al, 2018; Kim et al, 2019; Meng et al, 2019). Recombinant AMP MccJ25 can be large-scale produced by an MccJ25 expression vector and exhibit antimicrobial activity.…”

AP-64, Encoded by C5orf46, Exhibits Antimicrobial Activity against Gram-Negative Bacteria

“…Although AMPs are promising candidates for the development of novel antibiotics, it is difficult to produce sufficient quantities of these proteins because of their toxicity toward microbial expression hosts. Previously, the SUMO tag was used widely for the prokaryotic expression of recombinant proteins, such as abaecin (a 34 amino acid long antimicrobial peptide from honeybees) [17]. In this study, the SUMO tag was fused to the N terminus of AP-64 to express the SUMO-AP-64 fusion protein.…”

Sub3 inhibits Aspergillus flavus growth by disrupting mitochondrial energy metabolism, and has potential biocontrol during peanut storage

“…The plasma membrane plays important physiological roles in maintaining a homeostatic environment, exchanging materials, and transferring energy and information in the cell. [36][37][38] Once the integrity of the cell membrane is destroyed, substances inside the cell can leak, potentially resulting in cell death. 39,40 AMPs targeting cell membranes have been reported and researched for many years, and can be roughly divided into two categories; membrane destruction and non-membrane destruction.…”

“…The plasma membrane plays important physiological roles in maintaining a homeostatic environment, exchanging materials, and transferring energy and information in the cell 36–38 . Once the integrity of the cell membrane is destroyed, substances inside the cell can leak, potentially resulting in cell death 39,40 .…”