Expression systems for heterologous production of antimicrobial peptides

Parachin, Nádia Skorupa; Mulder, Kelly Cristina Leite; Viana, Antônio Américo Barbosa; Dias, Simoni Campos; Franco, Octávio L.

doi:10.1016/j.peptides.2012.09.020

Cited by 147 publications

(125 citation statements)

References 170 publications

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“…Antimicrobial peptides Antimicrobial peptides are also termed as host defense peptides which are present in all living organisms with an amino acid length of about 30 to 60 numbers. These peptides possess immunomodulatory and antimicrobial activity that can damage bacteria (by targeting cell membrane), virus and also fungus (Li et al, 2012;Parachin et al, 2012). Several of these antimicrobial peptides are identified and many were tested for their beneficial effects like growth promoter activity in poultry.…”

Section: Antimicrobials Of Plant Origin / Phytobioticsmentioning

confidence: 99%

Exploring alternatives to antibiotics as health promoting agents in poultry- a review

Yadav¹,

Kolluri²,

Gopi³

et al. 2016

JEBAS

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Poultry industry has undergone rapid growth during last three decades. For which even higher usage of antibiotics, both as growth promoters as well as therapeutic agents, has been adopted. However, due to the fear of resistance development in bacterial populations to antibiotics, presence of antibiotic residues in poultry products and increasing consumer demand for products free from antibiotic residues, search for alternatives that could replace antibiotics without causing loss to productivity or product quality has accelerated. Such alternatives in poultry include the use of organic acids, probiotic microorganisms, prebiotic substrates that benefit proliferation of beneficial bacterial populations or synbiotic (combinations of prebiotics and probiotics) ensuring better production and maintaining health of the birds. Others include vitamins and minerals, herbal drugs, plant extracts, phytobiotics and antimicrobial peptides. Probiotic organisms provides competition to pathogenic organisms for intestinal colonizing sites, reduce the diversion of nutrients for harmful microbes and the toxins produced by them and stimulates the immune systems. Similarly, prebiotic offers an alternative, as it alters the intestinal microbes and immune system to reduce colonization by pathogens and allows proliferation of beneficial microflora in the gut. Even using synbiotic is a better strategy for enhancing poultry production, however, more research is needed for selection of probiotic, prebiotics or synbiotics either alone or in combination that can result in the selection of strains capable of performing effectively in the gastrointestinal tract. The contents of this review will be useful for researchers to enrich their knowledge on alternatives of antibiotics in poultry birds without compromising performance of birds and bird welfare.

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Section: Antimicrobials Of Plant Origin / Phytobioticsmentioning

confidence: 99%

Exploring alternatives to antibiotics as health promoting agents in poultry- a review

Yadav¹,

Kolluri²,

Gopi³

et al. 2016

JEBAS

View full text Add to dashboard Cite

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“…Recombinant production in bacteria and yeasts expression systems provide an opportunity for industrial-scale production of AMPs [13]. Nevertheless, there are also some limitations.…”

Section: Introductionmentioning

confidence: 99%

Recombinant expression of LFchimera antimicrobial peptide in a plant-based expression system and its antimicrobial activity against clinical and phytopathogenic bacteria

Chahardoli

Fazeli

Niazi‎

et al. 2018

Biotechnology & Biotechnological Equipment

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Antimicrobial peptides (AMPs) with their broad and selective antimicrobial activity and lowered risk of resistance development in microbial populations are promining as a new alternative candidate to conventional antibiotics. Plant-based expression systems can be employed as cost-effective and high scale-up capacity production hosts for recombinant expression of AMPs. LFchimera is a chimerical peptide containing LFcin and LFampin antimicrobial peptides of bovine lactoferrin, and it has stronger bactericidal activity. Here, the LFchimera sequence was codon-optimized for tobacco and fused with endoplasmic reticulum retention signals along with a CaMV 35S promoter and then transferred by agrobacterium-mediated transformation. The integration and expression of the transgene in tobacco plants were confirmed by polymerase chain reaction (PCR) and RT-PCR, respectively. Recombinant production of the peptide was confirmed by sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE), and peptide functional activity was confirmed by the antibacterial evaluation of total protein extracts against various clinical and phytopathogenic bacteria. Finally, LFchimera peptide was partially purified using an affinity column, and the antibacterial activity was shown. Taken together, these results confirmed that tobacco can be utilized for the recombinant production of antimicrobial peptides such as LFchimera.

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“…So far, recombinant PSI domains have been produced only in E. coli (8)(9)(10). The low yields reported to be obtained clearly indicate that this bacterial expression system is only suboptimally suited for expression of PSIs, as well as other AMPs (15,16). In order to establish a more effective expression system for PSI domains (glycosylated and nonglycosylated) with improved production yields, we selected the GRAS yeast K. lactis as the tool of choice.…”

Section: Resultsmentioning

confidence: 99%

“…The development of low-cost production of AMPs has received much attention in recent years (16). Some advances were achieved in E. coli to overcome AMP toxicity by generation of fusion constructs to sequester AMPs in inclusion bodies.…”

Section: Discussionmentioning

confidence: 99%

“…Some advances were achieved in E. coli to overcome AMP toxicity by generation of fusion constructs to sequester AMPs in inclusion bodies. Subsequent protein-refolding steps and cleavage with an external protease yielded the desired product (15,16). The drawback of such an approach is multiple processing/purification steps, which take time and result in reduced protein yields and increased production costs.…”

Section: Discussionmentioning

confidence: 99%

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Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin

Curto¹,

Lufrano

Pinto

et al. 2014

Appl Environ Microbiol

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Typical plant aspartic protease zymogens comprise a characteristic and plant-specific insert (PSI). PSI domains can interact with membranes, and a role as a defensive weapon against pathogens has been proposed. However, the potential of PSIs as antimicrobial agents has not been fully investigated and explored yet due to problems in producing sufficient amounts of these domains in bacteria. Here, we report the development of an expression platform for the production of the PSI domain of cirsin in the generally regarded as safe (GRAS) yeast Kluyveromyces lactis. We successfully generated K. lactis transformants expressing and secreting significant amounts of correctly processed and glycosylated PSI, as well as its nonglycosylated mutant. A purification protocol with protein yields of ϳ4.0 mg/liter was established for both wild-type and nonglycosylated PSIs, which represents the highest reported yield for a nontagged PSI domain. Subsequent bioactivity assays targeting phytopathogenic fungi indicated that the PSI of cirsin is produced in a biologically active form in K. lactis and provided clear evidence for its antifungal activity. This yeast expression system thereby emerges as a promising production platform for further exploring the biotechnological potential of these plant saposin-like proteins.

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Expression systems for heterologous production of antimicrobial peptides

Cited by 147 publications

References 170 publications

Exploring alternatives to antibiotics as health promoting agents in poultry- a review

Exploring alternatives to antibiotics as health promoting agents in poultry- a review

Recombinant expression of LFchimera antimicrobial peptide in a plant-based expression system and its antimicrobial activity against clinical and phytopathogenic bacteria

Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin

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