Commercial utilization of bacteriocins: tackling challenges and exploring their potential as alternatives to antibiotics

ABSTRACT

The global surge in antibiotic resistance has accelerated the search for alternative antimicrobial strategies, with bacteriocins emerging as promising candidates. These ribosomally synthesized peptides exhibit potent, targeted antimicrobial activity against multidrug-resistant (MDR) pathogens and have shown efficacy in food preservation, pharmaceuticals, and clinical applications. Their specificity and rapid bactericidal action offer distinct advantages over conventional antibiotics, including a potentially lower risk of resistance development. Nonetheless, recent studies report resistance mechanisms such as membrane modifications, efflux pumps, and enzymatic degradation. Despite their potential, only a limited number of bacteriocins have been commercialized primarily for food applications with none yet approved for therapeutic use. Major barriers include low production yields, poor stability, and stringent regulatory challenges. This review presents a comprehensive overview of bacteriocins, including their classification, mechanisms of action, and commercial landscape. It further highlights current innovations aimed at overcoming translational bottlenecks, such as genetic engineering, nanotechnology-based delivery, and fermentation optimization. Ultimately, the review underscores the need for interdisciplinary research and harmonized regulatory frameworks to fully realize bacteriocins’ potential in addressing the global antibiotic resistance crisis.

Plain Language Summary

Bacteriocins are natural antimicrobial compounds made by bacteria that can kill harmful microbes without affecting beneficial ones. Because of their strong ability to fight bacteria, they are used as safe, natural preservatives in food products. However, turning bacteriocins into medicines is still difficult. Problems like low production, stability issues, and complicated regulations have slowed down their approval for medical use. Researchers are now improving how bacteriocins are made, using genetic tools and special technologies to make them more stable and easier to use. New methods like combining bacteriocins with nanoparticles or other delivery systems are helping to protect them until they reach their targets. Governments and industries are also working to create simpler approval processes. If these challenges are overcome, bacteriocins could become an important tool in hospitals and food industries to fight dangerous bacteria, especially those resistant to antibiotics. This review explains how bacteriocins work, their history, current commercial uses, the barriers holding them back, and new strategies that could make them a part of everyday healthcare and food safety systems.

1. Introduction

The increasing prevalence of antibiotic resistance has emerged as a global health crisis, necessitating the urgent development of alternative antimicrobial strategies. The overuse and misuse of conventional antibiotics have accelerated the rise of multidrug-resistant (MDR) pathogens, making bacterial infections more difficult to treat and contributing to increased morbidity, mortality, and healthcare costs. In response to these challenges, bacteriocins have gained significant attention as promising alternatives to traditional antibiotics due to their high specificity and rapid bactericidal action. Bacteriocins are ribosomally synthesized antimicrobial peptides produced by bacteria that typically exhibit narrow-spectrum activity against closely related strains [1]. Although initially thought to have lower resistance potential, recent studies show that resistance to bacteriocins can emerge through various mechanisms, including membrane alterations and efflux systems [2]. Their applications span various sectors, including food preservation, pharmaceuticals, and clinical therapeutics, making them valuable tools for addressing antibiotic resistance while ensuring microbial safety’s.

Bacteriocins have been extensively studied since the discovery of colicins in 1925 by André Gratia, a class of bacteriocins produced by Escherichia coli. This discovery-initiated decades of research, leading to the identification of diverse bacteriocins from both Gram-positive and Gram-negative bacteria as depicted in Figure 1 [3,4]. Bacteriocins exert their antimicrobial activity through multiple mechanisms, including membrane disruption, inhibition of cell wall synthesis, interference with protein synthesis, and DNA and RNA damage. Their narrow-spectrum activity enables them to selectively target bacterial competitors while minimizing disruption to beneficial microbiota, making them safer alternatives to broad-spectrum antibiotics [5]. Additionally, some bacteriocins exhibit synergistic effects when combined with antibiotics, enhancing their efficacy against resistant bacterial strains.

Figure 1.

Timeline of 99 years of bacteriocin discoveries.

This figure illustrates the chronological advancements in bacteriocin research over the past 99 years. It highlights key discoveries, from the initial identification of colicins to the modern exploration of bacteriocins in therapeutic and industrial applications.

Bacteriocins have already achieved commercial success as bio-preservatives in the food industry, enhancing food safety and extending shelf life. FDA-approved bacteriocins such as nisin and pediocin serve as key examples, effectively controlling spoilage organisms and foodborne pathogens. However, transitioning bacteriocins from food preservation to therapeutic applications presents significant challenges. Regulatory barriers, extensive clinical trial requirements, stability concerns, and production scalability issues hinder their widespread adoption. The FDA’s regulatory framework mandates rigorous safety and efficacy evaluations, making market entry both time-consuming and costly [6]. Furthermore, traditional fermentation-based bacteriocin production results in low yields, increasing production costs [7]. Additional challenges include enzymatic degradation and pH instability, which reduce their bioavailability in physiological environments.

To overcome these barriers, advancements in biotechnology and nanotechnology are being explored. Genetic engineering techniques, such as CRISPR-Cas, are improving bacteriocin production efficiency, while nanocarrier-based delivery systems (e.g., liposomes and polymeric nanoparticles) enhance stability and controlled release [8]. Synthetic biology approaches are also being employed to enhance bacteriocin discovery, production, and formulation.

This review provides a comprehensive analysis of bacteriocin commercialization, covering their classification, mechanisms of action, industrial applications, regulatory challenges, and production limitations. Special emphasis is placed on emerging technologies, including synthetic biology and nanotechnology, which are shaping the next generation of bacteriocin-based antimicrobial solutions. A multidisciplinary approach integrating biotechnology, regulatory science, and industrial optimization is essential for unlocking the full potential of bacteriocins. Addressing commercialization barriers will position bacteriocins as key players in the fight against antibiotic resistance, offering sustainable and effective alternatives to conventional antibiotics.

2. Classification of bacteriocins

Bacteriocins produced by Gram-positive and Gram-negative bacteria are classified based on structural composition, molecular characteristics, and producer organisms. Gram-positive bacteriocins are categorized into five distinct classes (Figure 2), while Gram-negative bacteriocins are classified into four types (Figure 3). These classifications offer critical insights for their potential applications in food safety, medical therapies, and biotechnology.

Figure 2.

Classification of bacteriocins produced by Gram-positive bacteria.

This schematic categorizes bacteriocins synthesized by Gram-positive bacteria into five major classes based on structural and functional features. It highlights lantibiotics, non-lantibiotics, large heat-labile proteins, post-translationally modified types, and multi-component bacteriocins, along with their representative producer organisms.

Figure 3.

Classification of bacteriocins produced by Gram-negative bacteria.

This figure illustrates the classification of bacteriocins produced by Gram-negative bacteria, grouped by their structural organization and mode of action. The major classes include colicins, colicin-like bacteriocins, phage-tail-like bacteriocins, and microcins.

2.1. Classification of bacteriocins by Gram-positive bacteria

Bacteriocins produced by Gram-positive bacteria are categorized into five major classes based on their structure, molecular features and producer organisms [9–13]. Class I bacteriocins, known as lantibiotics, contain unusual amino acids such as lanthionine and methyllanthionine resulting from extensive post-translational modifications. A well-known example is nisin, produced by Lactococcus lactis, and other producers include Streptococcus and Bacillus species. Class II bacteriocins are small, heat-stable peptides that do not contain lanthionine; they are further divided into subgroups including linear, cyclic, and thiopeptide variants. These are commonly produced by Lactobacillus, Pediococcus, and Enterococcus species. Class III bacteriocins are large proteins typically exceeding 30 kDa in size and often have complex quaternary structures; they are produced by organisms such as Bacillus cereus and Clostridium perfringens. Class IV bacteriocins are characterized by post-translational modifications such as glycosylation, which influence their stability and bioactivity. They are generally produced by Lactobacillus and certain Leuconostoc species. Finally, Class V bacteriocins are multi-component systems that require more than one protein for their antimicrobial functionality, often forming complexes to exhibit full activity, and are found in species like Streptococcus pneumoniae and Enterococcus faecalis.

2.2. Classification of bacteriocins by Gram-negative bacteria

Bacteriocins produced by Gram-negative bacteria are categorized into four major groups based on their structural characteristics and producer organisms [14,15]. Colicins are high-molecular-weight bacteriocins produced specifically by Escherichia coli and are encoded by plasmids. Colicin-like bacteriocins are synthesized by other Gram-negative species such as Klebsiella, Salmonella, and Pseudomonas, and exhibit structural and genetic similarities to colicins, though they are often species-specific [3,4]. Phage-tail-like bacteriocins, such as R-type pyocins from Pseudomonas aeruginosa, are protein complexes that structurally resemble bacteriophage tails. Microcins are low-molecular-weight, heat-stable peptides commonly produced by enteric bacteria like E. coli, Klebsiella, Shigella, and Salmonella [16]. These classifications based on molecular features help guide research into their applications in food safety, medical therapies, and biotechnological innovations, while also informing strategies to optimize production and stability [1].

3. Mode of action of bacteriocins

These peptides target essential cellular processes through distinct mechanisms of action. The primary modes of action include membrane disruption, inhibition of cell wall synthesis, interference with protein synthesis, and DNA and RNA damage. In Gram-positive bacteria, bacteriocins primarily disrupt the bacterial cell membrane by forming pores or binding to lipid II, thereby inhibiting peptidoglycan biosynthesis [7,17]. In Gram-negative bacteria, bacteriocins such as microcins and colicins employ diverse strategies, including pore formation in the membrane, inhibition of ribosomal functions, and disruption of DNA replication [18,19]. This section provides a detailed overview of the mechanisms through which different classes of bacteriocins exert their antimicrobial effects, emphasizing their potential as powerful alternatives to traditional antibiotics.

3.1. Mode of action of Gram-positive bacteriocins

Gram-positive bacteriocins primarily target the cell membrane of sensitive bacteria, forming pores that disrupt membrane integrity, ultimately leading to cell lysis. A well-known example is nisin, which binds to lipid II, an essential precursor in peptidoglycan biosynthesis, thereby inhibiting cell wall synthesis and facilitating pore formation [10,11]. Other Gram-positive bacteriocins, such as lactococcin and pediocin, also exhibit membrane-targeting activity.

Beyond membrane disruption, some Gram-positive bacteriocins inhibit protein synthesis by binding to ribosomal subunits, preventing the production of essential proteins. Additionally, certain bacteriocins interfere with metabolic pathways by inhibiting key enzymes involved in energy production, such as those participating in glycolysis. The specificity of these bacteriocins enables them to selectively target particular bacterial species, while the producing organism remains protected through immunity proteins that prevent bacteriocin activity against the host bacterium [16].

3.2. Mode of action of Gram-negative bacteriocins

3.2.1. Microcins – class 1

Class 1 microcins are small antimicrobial peptides produced by Gram-negative bacteria, including Escherichia coli and Klebsiella pneumoniae. These microcins exert their antimicrobial effects primarily by disrupting target cell membranes through pore formation, leading to ion leakage, loss of cellular integrity, and eventual bacterial cell death [14]. Class 1 microcins are also known for their ability to bind to specific receptors on the surface of susceptible bacteria, facilitating efficient uptake into the target cell. Once inside, they inhibit essential cellular processes, including protein synthesis, DNA replication, and enzyme activity. For instance, Microcin C inhibits aspartate tRNA synthetase, an enzyme crucial for protein synthesis, while Microcin B17 targets DNA gyrase, disrupting DNA replication [17–19].

3.2.2. Microcins – class 2

Class 2 microcins are unmodified peptides that exert antimicrobial activity by permeabilizing the bacterial membrane. These bacteriocins interact electrostatically with membrane components, causing disruption of membrane integrity and pore formation [15]. This results in ion imbalance and loss of energy potential, ultimately leading to bacterial cell death. Certain Class 2 microcins, such as Microcin E492, can penetrate the target cell membrane and interfere with essential cellular functions. Depending on the specific microcin, they may inhibit key processes such as protein synthesis or DNA replication [17,20]. Their ability to rapidly depolarize the membrane and compromise cellular integrity makes them potent antimicrobial agents against pathogenic bacteria [21,22].

3.2.3. Colicins

Colicins are large bacteriocins primarily produced by Escherichia coli, exhibiting antibacterial activity through a multi-domain mechanism. Their action begins with the receptor-binding domain, which recognizes and binds to specific receptors on the target bacterial surface, such as TolC or OmpF [23]. Once bound, the translocation domain facilitates the passage of colicins across the outer membrane, allowing them to enter the target cell. Inside the cell, the cytotoxic domain exerts its lethal effects through various mechanisms: Some colicins inhibit DNA replication by directly interacting with DNA. Others suppress protein synthesis by binding to ribosomal subunits [24]. Certain colicins disrupt membrane integrity by forming pores, leading to cell death through ion leakage and loss of cellular components.

The diverse mechanisms of colicins enable them to effectively target a broad range of Gram-negative bacteria, particularly within the Enterobacteriaceae family [16]. By understanding the diverse mechanisms through which bacteriocins exert their antimicrobial effects, researchers can optimize their applications in medicine, food preservation, and biotechnology. The ability of bacteriocins to selectively target specific bacterial populations while minimizing the risk of resistance development highlights their potential as next-generation antimicrobial agents.

4. Commercially available bio-preservatives

Bacteriocins, antibacterial peptides produced by lactic acid bacteria (LAB), serve as natural bio-preservatives. They have gained significant commercial recognition, particularly in the food industry, due to their Generally Recognized as Safe (GRAS) status by regulatory bodies such as the FDA and EFSA (Table 1) [25,26]. Among the most commercially utilized bacteriocins, nisin (Class I) and pediocin (Class II) have been successfully integrated into dairy, meat, and beverage products to prevent spoilage and microbial contamination. Additionally, sakacin and microcin, though not as widely commercialized, have demonstrated efficacy in preserving minimally processed and ready-to-eat foods, broadening their potential applications [27].

Table 1.

Commercially available bio-preservatives approved by FDA.

Bio-preservatives	Producing strain	Target organism	Commercial name	Applications	References
Nisin A	Lactococcus lactis	Listeria spp. Bacillus spp. Clostridium spp.	Nisaplin® Nisin A®	Dairy, meat, bakery, culinary products & beverages.	[1,22,26]
Nisin Z	Lactococcus lactis	Listeria spp. Clostridium spp. Bacillus cereus	Nisin Z®	Dairy, meat, bakery products, delicacies & beverages.
Nisin	Lactococcus lactis	Clostridium botulinum, Listeria monocytogens	Chrisin ®	Meat, sausages & spore forming bacteria in cheese.
Natamycin	Streptomyces natalensis	Yeast & molds	Natamax®	Cheese, fresh dairy products, beverages and processed meat.	[35,36]
Pediocin	Pediococcus acidilactici	Listeria monocytogens	ALTA® 2351,2341& Fargo 23®	Meat products	[9,37]
Sakacin	Lactobacillus sakei strain Lb674, LT673	Listeria monocytogens	Bactoferm®	Meat products	[41]
Microcin	E. coli & Klesiella pneunomia	Listeria monocytogens	Microcin®	Meat products	[14,40,41]

This table lists bacteriocin-based bio-preservatives that have received FDA approval for use in food products. It includes details on the producing strains, target organisms, commercial names, applications in food industries, and relevant references. These bio-preservatives play a crucial role in enhancing food safety by inhibiting pathogenic and spoilage bacteria.

Despite these successes, the global market for bacteriocins remains underdeveloped compared to synthetic preservatives and conventional antibiotics used in food processing. Factors limiting industrial adoption include high production costs, limited fermentation scalability, low consumer awareness, and the potential gene transfer of bacteriocin resistance via conjugative plasmids, which may impact long-term efficacy [16]. Recent studies have demonstrated the feasibility of conjugation-based antimicrobial systems, where bacteriocin genes are transferred via conjugative plasmids, highlighting both biosafety implications and innovative applications in microbial control strategies [2]. However, major food manufacturers are now exploring synergistic combinations of bacteriocins with natural preservatives such as organic acids and essential oils to enhance food shelf-life and cost-effectiveness, creating new opportunities for market expansion [28,29].

Beyond food preservation, bacteriocins are gaining attention in pharmaceutical and biotechnology sectors. Ongoing research is investigating bacteriocin-based antimicrobial coatings for medical devices, probiotic formulations, and wound care dressings. While these applications are still in early-phase clinical trials, they reflect an industry shift toward sustainable antimicrobial alternatives. However, successful commercialization in medical applications will depend on cost-effective production innovations, regulatory approvals, and optimization of stability and bioavailability [30–33].

4.1. Nisin

Nisin, a bactericidal peptide produced by Lactococcus lactis, is a prominent member of Class I bacteriocins [34]. It exists in four natural variants: Nisin A, Z, Q, and U. Among these, Nisin A and Z are the most commercially available due to their potent antimicrobial properties [26]. Nisin exerts antimicrobial activity primarily by inhibiting peptidoglycan biosynthesis, disrupting bacterial cell wall formation, which leads to cell lysis and death [7]. Additionally, nisin binds to lipid II, a key intermediate in cell wall synthesis, facilitating pore formation in the bacterial membrane. This pore formation results in ion influx and solute leakage, ultimately causing bacterial death [22]. However, nisin resistance can develop through multiple mechanisms, including membrane receptor alterations, efflux pumps, and in some cases, plasmid-mediated gene transfer. Notably, membrane-associated resistance appears to be more prevalent than conjugative transfer in many settings [2].

4.2. Natamycin

Natamycin, also known as pimaricin, is a naturally occurring antifungal peptide produced by Streptomyces natalensis. It is widely used as a preservative in dairy products, meat, juices, and wine [35]. As a macrolide and polyene antifungal agent, natamycin inhibits fungal growth by targeting ergosterol, an essential component of the fungal cell membrane. By binding to ergosterol, natamycin disrupts membrane integrity, increasing permeability and ultimately leading to cell death. Additionally, natamycin impedes amino acid and glucose transport proteins, reducing nutrient uptake, further contributing to fungal cell death [36]. While natamycin is highly effective against fungal contamination, resistance may develop through mechanisms such as alterations in ergosterol biosynthesis, efflux pump activation, or membrane composition changes. Though less frequently, resistance genes may also be disseminated via horizontal gene transfer mechanisms [2].

4.3. Pediocin

Pediocins, antimicrobial proteins produced by Pediococcus acidilactici, belong to Class II bacteriocins (non-lantibiotics) [9]. They exhibit a bactericidal mode of action by targeting the bacterial cytoplasmic membrane. Pediocins first bind to specific receptors on the bacterial membrane, allowing them to integrate into the membrane, disrupting its integrity. This leads to pore formation, which results in ion leakage and bacterial cell death [37]. Pediocins are highly stable under various pH and temperature conditions, making them suitable for diverse food preservation applications. While horizontal gene transfer is a documented concern, bacteriocin resistance more frequently arises through mutations in membrane receptors or regulatory pathways that affect peptide uptake and pore formation [2].

4.4. Sakacin

Sakacin A, a Class IIa bacteriocin produced by Lactobacillus sakei, is a 41-amino acid peptide known for its strong antimicrobial activity against Listeria species [38]. The bactericidal mechanism of Sakacin A involves permeabilizing the target bacterial membrane, leading to pore formation and subsequent cell death. Due to its potent antilisterial activity, sakacin A has significant potential as a bio-preservative. Its effectiveness at low concentrations makes it economically viable for industrial applications, while its stability across various pH levels and temperatures enhances its usability in different food preservation environments. However, resistance can emerge through various mechanisms, including receptor modification and gene transfer, which may compromise its long-term effectiveness [39].

4.5. Microcin

Microcins, low molecular weight bacteriocins produced by Gram-negative bacteria such as Escherichia coli and Klebsiella pneumoniae, exhibit diverse mechanisms of action [14,40]. Microcins employ various antimicrobial strategies, including membrane pore formation, inhibition of protein synthesis, and disruption of DNA replication. Microcin V disrupts the bacterial membrane, leading to ion leakage and cell death. Microcin C inhibits aspartate tRNA synthetase, blocking protein synthesis. Additionally, Microcin B17 interferes with DNA gyrase, causing double-strand breaks and leading to bacterial cell death [41]. These diverse mechanisms highlight microcins’ versatility as antimicrobial agents, making them promising candidates for food and medical applications. However, resistance can develop through impaired uptake mechanisms, efflux pumps, or membrane modifications. Though plasmid-mediated gene transfer has been observed, it is not the primary mode of resistance [2].

5. Progress and challenges of bacteriocins in clinical and pre-clinical trials

Bacteriocins produced by bacteria, have demonstrated significant potential as therapeutic agents, particularly in oncology and antimicrobial therapies. Notably, nisin and pediocin have shown promise in antimicrobial applications, while azurin-derived peptides and epidermicins are being explored for their potential anticancer properties. Despite their therapeutic potential, only a limited number of bacteriocins have progressed to clinical trials, and none have received regulatory approval as commercial drugs [42].

5.1. Bacteriocins in clinical and pre-clinical development

5.1.1. Bacteriocins in human clinical trials: advancements and limitations

Bacteriocins have emerged as promising antimicrobial agents, with several candidates progressing to human clinical trials. Despite their potent antimicrobial activity, challenges such as trial progression, regulatory approval, biosafety validation, and formulation stability hinder their clinical translation [43]. The clinical development landscape of bacteriocins primarily focuses on bacterial infections and oncological applications. Notably, Nisin ZP has demonstrated potential antitumor properties in trials for non-metastatic oral squamous cell carcinoma [44–46]. Additionally, OG716, a derivative of mutacin, has exhibited safety and effectiveness in Clostridioides difficile infection models [43]. A comprehensive summary of bacteriocins currently undergoing clinical evaluation, their status, and associated challenges is provided in Table 2.

Table 2.

Bacteriocins in clinical development: candidates and current progress.

Bacteriocin	Source organism	Year	Status	Current progress and challenges	References
Nisin ZP	Lactococcus lactis	2023	Phase I/II	Evaluated for non-metastatic oral squamous cell carcinoma; promising antitumor properties.	[43–46]
OG716	Mutacin derivative	2020	Phase I	Demonstrated safety and effectiveness in C. difficile infection models.

This table provides an overview of bacteriocins that have progressed to clinical trials, highlighting their source organisms, year of clinical entry, current trial status, and key findings. It includes candidates such as Nisin ZP and OG716 outlining their therapeutic potential and challenges in clinical translation.

5.1.2. Bacteriocins in pre-clinical development: potential and hurdles

Several bacteriocins are undergoing preclinical development, demonstrating promising antimicrobial properties against various pathogens. However, their transition to clinical applications faces multiple challenges, including stability issues, high production costs, regulatory hurdles, and biosafety concerns. These obstacles arise from factors such as enzymatic degradation, complex purification methods, and uncertainties in regulatory classification [43]. Addressing these limitations through optimized production strategies, clearer regulatory frameworks, and extensive in vivo studies is crucial for advancing bacteriocin-based therapies [47]. Table 3 provides an overview of key bacteriocins currently undergoing preclinical investigation, along with their sources, development status, and associated challenges.

Table 3.

Preclinical bacteriocins: development status and key challenges.

Bacteriocin	Source organism	Year	Status	Current progress and challenges	References
MU1140	Streptococcus mutans	2015	Pre-Clinical	Stability issues and high production costs hinder development.	[43–46]
Lacticin 3147	Lactococcus lactis	2018	Pre-Clinical	Regulatory hurdles and safety concerns delay progression.
Pediocin PA-1	Pediococcus acidilactici	2015	Pre-Clinical	Further in vivo studies required to assess safety and efficacy.
EntV	Enterococcus faecalis	2020	Pre-Clinical	Investigated for anti-Candida albicans activity.
Duramycin	Lantibiotic	2020	Pre-Clinical	Under evaluation for cystic fibrosis therapy.

This table summarizes bacteriocins that are in preclinical research stages, focusing on their source organisms, year of investigation, development status, and associated challenges. It includes promising candidates like MU1140, Lacticin 3147, Pediocin PA-1, EntV, and Duramycin, detailing their potential applications and barriers to commercialization.

5.1.3. Developmental pipeline and challenges

The clinical translation of bacteriocins follows a structured pipeline that includes discovery, preclinical studies, and clinical trials, as illustrated in Figure 4 [43]. The process begins with environmental sampling and bioinformatic analysis to identify bacteriocins with potent antimicrobial properties [42]. Identified candidates undergo in vitro and in vivo screening to assess their efficacy, toxicity, and biosafety before advancing to clinical evaluation.

Figure 4.

Bacteriocin Development Pipeline: From Discovery to Clinical Trials. A schematic representation of the stages involved in bacteriocin development, from initial discovery and bioinformatic screening to preclinical and clinical trials. The figure highlights key challenges and regulatory steps required for clinical translation (adapted from [43]).

Despite their potential, several critical challenges hinder the widespread clinical adoption of bacteriocins. Toxicity and biosafety concerns remain significant limitations. While bacteriocins have been shown to modulate immune responses, regulate inflammatory pathways, and reduce infection-associated damage, the majority of in vivo studies primarily focus on antimicrobial efficacy without rigorous toxicity assessments [15]. Notably, 47.4% of murine model studies assess only antimicrobial activity, while 20% of alternative models lack toxicity evaluations entirely [43]. This gap in biosafety validation, as outlined in Table 3, represents a major bottleneck in the transition of bacteriocins into human therapeutics.

5.2. FDA regulatory framework and challenges in commercializing bacteriocins

Bacteriocins have been extensively explored for food preservation and therapeutic applications against foodborne pathogens. The U.S. Food and Drug Administration (FDA) regulates bacteriocins through two primary pathways: (1) Generally Recognized as Safe (GRAS) program for food applications, (2) New Drug Application (NDA) process for therapeutic use [48].

5.2.1. Bacteriocins approved for food use

Bacteriocins such as nisin and colicins have been granted GRAS status for specific food applications, meaning they are considered safe under intended conditions of use. The FDA’s GRAS notices relevant to bacteriocins include: GRAS Notice No. 593 - Colicin preparations for antimicrobial use on fresh or processed vegetables and fruits at 1–10 mg/kg [49]. GRAS Notice No. 676 - Colicin preparations for meats (beef, pork, lamb, veal) at 1–10 mg/kg [50]. GRAS Notice No. 824 - Bacteriocin preparations targeting Salmonella on meat, poultry, fish, and egg products at 0.1–3 mg/kg [51]. These approvals allow bacteriocins to be incorporated into food products without requiring premarket approval, provided they meet safety and efficacy standards.

5.2.2. Bacteriocins as therapeutic drugs

For bacteriocins intended as therapeutic agents, they must undergo rigorous clinical testing under the NDA pathway, which involves: Preclinical studies (laboratory and animal testing to evaluate safety and efficacy). Clinical trials (multiple phases assessing dosage, side effects, and therapeutic effectiveness). FDA review and approval, requiring submission of an NDA with comprehensive pharmacokinetics, toxicity, and manufacturing data [52]. Currently, no bacteriocin has received full FDA approval as a therapeutic drug, primarily due to challenges in demonstrating clinical efficacy and stability [42].

5.2.3. Challenges in bacteriocin commercialization

Several regulatory challenges hinder the commercialization of bacteriocins. The dual classification issue where bacteriocins can be GRAS-approved food preservatives but require extensive clinical trials for drug approval poses a significant regulatory hurdle, as the latter process is costly and time-consuming [53]. Additional commercialization barriers include: Toxicity and safety concerns, requiring thorough toxicological evaluations to address risks such as allergenicity, immunogenicity, and unintended antimicrobial resistance [43]. Large-scale production constraints, requiring adherence to Good Manufacturing Practices (GMP) to ensure batch-to-batch reproducibility [54]. Stability and formulation issues, as bacteriocins degrade rapidly in certain environments, necessitating innovative delivery mechanisms [55].

Bacteriocins offer promising potential as natural preservatives and therapeutic agents, but regulatory and commercialization challenges remain significant hurdles. The Limited Population Pathway for Antibacterial and Antifungal Drugs (LPAD) and FDA guidelines on antibacterial therapies provide structured pathways for drug development, particularly for urgent medical needs [56]. Future biotechnological advancements and regulatory innovations will be crucial in streamlining approval processes and expanding bacteriocin applications in food preservation and medicine [32,57].

6. Strategies for overcoming commercialization barriers of bacteriocins

The commercialization of bacteriocins as viable antimicrobial agents faces multiple challenges, including low production yields, stability concerns, regulatory complexities, and limited consumer awareness. To facilitate their broader adoption in food preservation, pharmaceuticals, and healthcare applications, various strategic and technological advancements are being implemented. Figure 5 provides a comprehensive roadmap to bacteriocin commercialization, outlining key stages from strain selection and production optimization to regulatory approval and market entry. Each phase highlights critical technical, regulatory, and industrial considerations necessary for successful large-scale application.

Figure 5.

Strategic roadmap for bacteriocin commercialization.

This flowchart outlines the essential steps for bringing bacteriocins to market, including production optimization, regulatory approval, and industry adoption. It emphasizes the role of biotechnology, regulatory science, and commercialization strategies in ensuring the successful integration of bacteriocins into food safety and medical applications.

6.1. Scaling up production with genetic engineering

A major bottleneck in bacteriocin commercialization is low production yield in native bacterial strains. Advances in synthetic biology and metabolic engineering offer promising solutions to enhance bacteriocin production. Techniques such as CRISPR-Cas, recombinant DNA technology, and engineered microbial expression systems (e.g., Escherichia coli and Lactococcus lactis) have demonstrated the ability to increase bacteriocin yield while reducing production costs. Additionally, metagenomic screening and high-throughput sequencing facilitate the discovery of novel bacteriocins with enhanced antimicrobial properties, further expanding the potential applications of these bioactive [12,58].

6.2. Cost-effective fermentation and alternative substrates

Traditional fermentation-based bacteriocin production relies on complex growth media, making the process cost-intensive. Optimizing fermentation parameters, including pH control, oxygenation, and nutrient supplementation, has been explored as a method to enhance yield while reducing costs. Additionally, utilizing agricultural byproducts such as molasses, whey, and plant-based extracts like soursop leaf as alternative low-cost substrates presents an economically viable strategy for large-scale bacteriocin manufacturing [30,59,60].

6.3. Advanced formulation and delivery technologies

Bacteriocins often suffer from stability issues, particularly under extreme temperature and pH variations, limiting their effectiveness in food and pharmaceutical applications. To address this, various encapsulation technologies, including liposomes, polymeric nanoparticles, and hydrogel-based carriers, have been developed to enhance bacteriocin stability and enable controlled release [61,62]. Additionally, spray-drying and freeze-drying techniques have been employed to extend the shelf life of bacteriocins, making them more suitable for large-scale commercial distribution [63].

6.4. Regulatory streamlining and consumer awareness

Regulatory approval remains a significant challenge due to the lack of standardized global frameworks, with different regions implementing varying safety and efficacy requirements. The FDA in the United States mandates rigorous clinical and toxicological evaluations, while the EFSA in Europe emphasizes comprehensive risk assessments for food safety applications. Additionally, WHO guidelines focus on the global public health impact, creating inconsistencies in approval processes across jurisdictions. These regulatory discrepancies complicate market entry, highlighting the need for a harmonized international regulatory approach to facilitate bacteriocin commercialization [57].

Early engagement with regulatory agencies such as the FDA, EFSA, and WHO, combined with comprehensive safety evaluations, can streamline the approval process. Developing a unified international regulatory framework for bacteriocin approval would help ensure regulatory consistency, allowing for faster market entry in food safety and healthcare industries [32,64]. Establishing collaborations between regulatory bodies and biotech firms will be critical in accelerating bacteriocin approval pathways, ensuring faster and broader adoption across multiple industries. Additionally, increasing consumer awareness through public outreach campaigns and industry collaborations will enhance market acceptance of bacteriocins as safe and effective antimicrobial alternatives [65].

6.5. Industry collaboration and investment strategies

Strong partnerships between academia, industry, and biotechnology firms are essential for accelerating bacteriocin commercialization [66]. Investments in bacteriocin-based products, probiotic formulations, and antimicrobial coatings by biotech startups and pharmaceutical companies have increased significantly [67]. Integrating bacteriocins into existing food and pharmaceutical pipelines will allow for cost-effective scaling and broader industrial adoption [68].

6.6. Expanding market applications

Beyond food preservation, bacteriocins show immense promise in biomedical applications, including antimicrobial coatings for medical devices, probiotic therapeutics, and novel drug formulations. Additionally, combining bacteriocins with traditional antibiotics in synergistic formulations may enhance efficacy against multidrug-resistant (MDR) pathogens, reducing antibiotic dosages and associated side effects [69–71]. Expanding bacteriocin applications beyond food safety into healthcare and pharmaceutical sectors will significantly improve their commercial viability and market potential.

7. Conclusion

Bacteriocins are emerging as promising natural antimicrobial peptides with significant potential in food preservation, medicine, and biotechnology. Their high specificity, potency against multidrug-resistant pathogens, and Generally Recognized as Safe (GRAS) status underscore their potential as complementary tools to conventional antibiotics and chemical preservatives. However, their widespread adoption remains hindered by challenges such as low production yields, stability issues, regulatory barriers, and limited consumer awareness. Recent advancements in genetic engineering, fermentation technology, and nanotechnology are actively addressing these limitations, enhancing bacteriocin scalability, bioavailability, and commercial viability.

To successfully integrate bacteriocins into mainstream applications, regulatory streamlining, industry collaborations, and consumer education will be essential. Continued innovation in formulation and delivery methods, combined with interdisciplinary efforts among researchers, biotechnologists, policymakers, and industry stakeholders, will be key to unlocking their full potential. With the escalating threat of antibiotic resistance, bacteriocins are poised to serve as a next-generation antimicrobial strategy. Sustained investment in research, regulatory alignment, and biotechnological innovation will be essential to translate laboratory success into clinical and industrial impact. If these barriers are overcome, bacteriocins have the potential to revolutionize both food safety and antimicrobial therapy, offering a sustainable, targeted, and globally relevant solution to one of the most pressing challenges in modern medicine.

8. Future perspectives

The future of bacteriocins as next-generation antimicrobial agents lies in overcoming key barriers related to scalability, regulatory approval, and clinical validation. Continued advances in synthetic biology, metabolic engineering, and fermentation optimization are poised to enhance production yields and lower manufacturing costs, making bacteriocins more commercially viable. In addition, nanotechnology-based delivery systems and novel formulation strategies will be instrumental in improving bacteriocin stability and bioavailability, expanding their applicability across food, pharmaceutical, and medical device industries. Strengthening interdisciplinary collaborations between microbiologists, bioengineers, regulatory experts, and industrial partners will be crucial to drive innovation and streamline commercialization pathways.

Equally important will be addressing biosafety concerns through rigorous toxicological and immunological evaluations in both preclinical and clinical studies. Regulatory harmonization across global jurisdictions and the establishment of standardized guidelines specific to bacteriocins will further facilitate market entry and adoption. Expanding research into the synergistic use of bacteriocins with conventional antibiotics and alternative antimicrobials could also offer promising strategies to combat multidrug-resistant pathogens. As global interest in sustainable and natural antimicrobial solutions grows, bacteriocins are well positioned to play a transformative role in ensuring food safety, enhancing therapeutic options, and contributing to the broader fight against antimicrobial resistance in the coming decades.

Funding Statement

This paper was not funded.

Article highlights

• Bacteriocins are ribosomally synthesized antimicrobial peptides with potent activity against multidrug-resistant (MDR) pathogens, positioning them as promising natural alternatives to conventional antibiotics and chemical preservatives.

• Commercial utilization of bacteriocins is well-established in the food industry, particularly as bio-preservatives, with notable examples including nisin and pediocin.

• Despite their efficacy, the therapeutic application of bacteriocins faces key limitations, such as low production yield, poor stability, narrow activity spectrum, and regulatory complexity.

• Recent innovations in genetic engineering, synthetic biology, and heterologous expression systems have enhanced bacteriocin biosynthesis, spectrum modification, and host compatibility.

• Fermentation and process optimization strategies, including media design and fed-batch cultivation, are being explored to scale up production cost-effectively.

• Nanotechnology-based delivery systems, such as nanoemulsions, liposomes, and biopolymeric carriers, have improved the stability, targeting, and controlled release of bacteriocins in both food and therapeutic contexts.

• Synergistic approaches combining bacteriocins with antibiotics or plant-derived compounds are being investigated to overcome resistance mechanisms and broaden antibacterial efficacy.

• Regulatory harmonization across global agencies remains a critical challenge, with a need for clear frameworks to facilitate bacteriocin approval for clinical use.

• Interdisciplinary collaboration among microbiologists, bioengineers, food technologists, and clinicians is essential to bridge the gap between lab-scale research and commercial or clinical deployment.

• With continued research and translational focus, bacteriocins offer immense potential not only to enhance food safety but also to contribute to next-generation antimicrobial therapies against MDR pathogens.

Disclosure statement

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Writing disclosure

No writing assistance was utilized in the production of this manuscript.

Authors contributions

Author Susai Arul Selvan led the review process, conceptualizing the framework, conducting literature searches, analyzing data, drafting the manuscript, and designing and creating tables and diagrams. Vihang Vivek Ghalsasi designed the conceptual framework and provided critical insights, while Anwesha Bandyopadhyay contributed to the overseeing revisions. Tarun Pal provided expert review and guidance.