Heterologous expression of abaecin peptide from Apis mellifera in Pichia pastoris

Luiz, Denis Prudencio; Almeida, Juliana Franco; Goulart, Luíz Ricardo; Nicolau-Júnior, Nilson; Ueira-Vieira, Carlos

doi:10.1186/s12934-017-0689-6

Cited by 27 publications

(12 citation statements)

References 41 publications

(37 reference statements)

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“…For high production of large peptides, biological systems such as bacteria and yeast have been preferably used as production platforms [7][8][9][10][11][12][13]. Although these biological systems don't need expensive active pharmaceutical ingredients (APIs) and toxic chemical solvents, there are still some issues remaining, such as the toxicity of the expressed AMPs to host cells, expensive purification, and low yield.…”

Section: Introductionmentioning

confidence: 99%

“…As a proline-rich, non-glycosylated antimicrobial peptide [27], abaecin has a broad spectrum of bacteriolytic activity [30] and shows increased inhibitory effects on bacterial growth when treated with pore-forming peptides, such as cecropin A [13,31], stomoxyn, and hymenoptaecin. The 34-aa-long cationic peptide contains 10 prolines (29%) with no cysteine residue, and the uniformed distribution of the proline residues through the entire peptide prevents the α-helical conformation [12,27].…”

Section: Introductionmentioning

confidence: 99%

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

Kim

Song

et al. 2019

BMC Biotechnol

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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.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Kim

Song

et al. 2019

BMC Biotechnol

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“…Li et al expressed the peptide CGA-N46 in Bacillus subtilis DB1342 [17]. Luiz et al produced the abaecin peptide [18] and Li et al expressed the antimicrobial peptide fowlicidin-2, both in Pichia pastoris [19]. In the current study, we expressed recombinant AC-1 using an E. coli prokaryotic expression system, and the AC-1 obtained by enterokinase digestion exhibited similar antimicrobial activity to chemically synthesized AC-1.…”

Section: Discussionmentioning

confidence: 68%

Identification and characteristics of a novel cecropin from the armyworm, Mythimna separata

et al. 2020

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Background: The recent emergence of antibiotic-resistant strains of bacteria has increased the need to develop effective alternatives to antibiotics. Antimicrobial peptides have been considered as a promising product with several advantages. Results: In this present study, we identified a novel cecropin from the armyworm, Mythimna separata (armyworm cecropin 1, AC-1) by transcriptome sequencing and multi-sequence alignment analysis. The AC-1 precursor comprised 63 amino acid residues, containing a conserved cleavage site of the signal peptide, Ala 23-Pro 24 , while the mature AC-1 included 39 amino acid residues. Chemically synthesized AC-1 exhibited low hemolytic activity against chicken red blood cells, low cytotoxicity against swine testis cells, and effective antimicrobial activity against Salmonella, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. Its antimicrobial activity against Salmonella remained after incubation for 1 h at 100°C or in 250 mM NaCl, KCl, or MgCl 2 solution, implying good thermal-and salt-resistant stabilities. The bactericidal effect of AC-1 on E. coli gradually increased with increasing AC-1 concentration, resulting in deformation, severe edema, cytolysis, cell membrane damage, and reducing intracellular electron density. Additionally, recombinant AC-1 protein expressed in E. coli was digested by enterokinase protease to obtain AC-1, which showed similar antimicrobial activity against E. coli to chemically synthesized AC-1. Conclusions: This study identified a novel antimicrobial peptide that may represent a potential alternative to antibiotics.

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“…Li et al expressed the peptide CGA-N46 in Bacillus subtilis DB1342 [17]. Luiz et al produced the abaecin peptide [18] and Li et al expressed the antimicrobial peptide fowlicidin-2 both in Pichia pastoris [19]. In this study, we evaluated the antimicrobial, hemolytic, and cytotoxic activities, and the thermal-and salt-resistant stabilities of chemically synthesized AC-1.…”

Section: Discussionmentioning

confidence: 99%

Identification and characteristics of a novel cecropin from the armyworm, Mythimna separata

Lian¹,

Zhang

Liang

et al. 2019

Preprint

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Background: The recent emergence of antibiotic-resistant strains of bacteria has increased the need to develop effective alternatives to antibiotics. Antimicrobial peptides have been considered as a promising product with several advantages.Results: In this present study, we identified a novel cecropin from the armyworm, Mythimna separata (armyworm cecropin 1, AC-1) by transcriptome sequencing and multi-sequence alignment analysis. The AC-1 precursor comprised 63 amino acid residues, containing a conserved cleavage site of the signal peptide, Ala23-Pro24, while the mature AC-1 included 39 amino acid residues. Chemically-synthesized AC-1 exhibited low hemolytic activity against chicken red blood cells and low cytotoxicity against swine testis cells, but effective antimicrobial activity against Salmonella, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. Its antimicrobial activity against Salmonella remained after incubation for 1 h at 100 °C or in 250 mM NaCl solution, implying good thermal- and salt-resistant stabilities.Conclusions: This study identified a novel antimicrobial peptide that may represent a potential alternative to antibiotics.

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Heterologous expression of abaecin peptide from Apis mellifera in Pichia pastoris

Cited by 27 publications

References 41 publications

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

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

Identification and characteristics of a novel cecropin from the armyworm, Mythimna separata

Identification and characteristics of a novel cecropin from the armyworm, Mythimna separata

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