Enzymatic Degradation Research Articles

Fibroin-Hybrid Systems: Current Advances in Biomedical Applications

Fibroin, a protein extracted from silk, offers advantageous properties such as non-immunogenicity, biocompatibility, and ease of surface modification, which have been widely utilized for a variety of biomedical applications. However, in vivo studies have revealed critical challenges, including rapid enzymatic degradation and limited stability. To widen the scope of this natural biomacromolecule, the grafting of polymers onto the protein surface has been advanced as a platform to enhance protein stability and develop smart conjugates. This review article brings into focus applications of fibroin-hybrid systems prepared using chemical modification of the protein with polymers and inorganic compounds. A selection of recent preclinical evaluations of these hybrids is included to highlight the significance of this approach.

Impact of Enzymatic Degradation Treatment on Physicochemical Properties, Antioxidant Capacity, and Prebiotic Activity of Lilium Polysaccharides

In order to overcome the bioavailability limitation of Lilium polysaccharide (LPS) caused by its high molecular weight and complex structure, two low-molecular-weight degraded polysaccharides, namely G-LPS(8) and G-LPS(16), were prepared through enzymatic degradation. The molecular weight of LPS was significantly reduced by enzymolysis, leading to increased exposure of internal functional groups and altering the molar ratio of its constituent monosaccharides. The results of antioxidant experiments showed that enzymatic hydrolysis had the potential to enhance the antioxidant performance of LPS. In vitro fermentation experiments revealed that LPS and its derivatives exerted different prebiotic effects on intestinal microbial communities. Specifically, LPS mainly inhibited the growth of harmful bacteria such as Fusobacterium, while G-LPS(8) and G-LPS(16) tended to promote the growth of beneficial bacteria like Megamonas, Bacteroides, and Parabacteroides. Metabolomic analysis revealed that LPSs with varying molecular weights exerted comparable promoting effects on multiple amino acid and carbohydrate metabolic pathways. Importantly, with the reduction in molecular weight, G-LPS(16) also particularly stimulated sphingolipid metabolism, nucleotide metabolism, as well as ascorbic acid and uronic acid metabolism, leading to the significant increase in specific metabolites such as sphingosine. Therefore, this study suggests that properly degraded LPS components have greater potential as a prebiotic for improving gut health.

The Evolution of Antimicrobial Resistance in Acinetobacter baumannii and New Strategies to Fight It

Acinetobacter baumannii is considered one of the prioritized ESKAPE microorganisms for the research and development of novel treatments by the World Health Organization, especially because of its remarkable persistence and drug resistance. In this review, we describe how this can be acquired by the enzymatic degradation of antibiotics, target site modification, altered membrane permeability, multidrug efflux pumps, and their ability to form biofilms. Also, the evolution of drug resistance in A. baumannii, which is mainly driven by mobile genetic elements, is reported, with particular reference to plasmid-associated resistance, resistance islands, and insertion sequences. Finally, an overview of existing, new, and alternative therapies is provided.

Cryo-SEM and large volume FIB-SEM of Arabidopsis cotyledons: Degradation of lipid bodies, biogenesis of glyoxysomes and reorganisation of organelles during germination.

Until recently, the lack of three-dimensional visualisation of whole cells at the electron microscopic (EM) level has led to a significant gap in our understanding of the interaction of cellular organelles and their interconnection. This is particularly true with regard to the role of the endoplasmic reticulum (ER). In this study, we perform three-dimensional reconstructions of serial FIB/SEM stacks and anaglyphs derived from volume rendering, cryo-scanning electron microscopy (cryo-SEM) and state-of-the-art electron microscopy immobilisation and imaging techniques. The results show that glyoxysomes are formed de novo in large numbers and in characteristic clusters on the ER upon germination in mesophyll cells of Arabidopsis cotyledons. The degradation of lipid bodies during germination occurs not only via the ER, which enlarges by taking up polar lipids resulting from enzymatic degradation by lipases, but also via glyoxysomes, which engulf lipid bodies. Dictyosomal (Golgi-derived) vesicles, which fuse with glyoxysomes or their precursors, also appear to be involved in the differentiation of glyoxysomes from segments of the ER. The formation of the central vacuole is the result of the fusion of protein storage vacuoles (protein bodies), which become complex three-dimensional structures during germination. Our observations also suggest that the vacuole plays a role in the degradation of glyoxysomes. The evidence provided in three dimensions shows that the endoplasmic reticulum plays a central role in the biogenesis and degradation of lipid bodies, the ontogeny of glyoxysomes and the development of plastids in the mesophyll cells of Arabidopsis cotyledons.

Identification and Characterization of a New Thermophilic κ-Carrageenan Sulfatase.

Carrageenans are sulfated polysaccharides found in the cell wall of certain red seaweeds. They are widely used in the food industry for their gelling and stabilizing properties. In nature, carrageenans undergo enzymatic modification and degradation by marine organisms. Characterizing these enzymes is crucial for understanding carrageenan utilization and may eventually enable the development of targeted processes to modify carrageenans for industrial applications. In our study, we characterized a κ-carrageenan sulfatase, AMOR_S1_16A, belonging to the sulfatase S1_16 subfamily, which selectively desulfates the nonreducing end galactoses of κ-carrageenan oligomers in an exomode. Notably, AMOR_S1_16A represents the first κ-carrageenan sulfatase within the S1_16 subfamily and exhibits a novel enzymatic activity. This study provides further understanding of the substrate specificity and characteristics of the S1_16 subfamily. Moreover, this research highlights that many processes and enzymes remain to be discovered to fully understand carrageenan utilization pathways and to develop enzymatic processes for carrageenan modification and processing.

Autocatalytic Ceria Nanoparticle-Embedded Tilapia Collagen Hydrogels as Enhanced Antioxidative and Long-Lasting Dermal Fillers for Photoaged Skin.

Excessive reactive oxygen species (ROS) generated by ultraviolet (UV) irradiation significantly contribute to photoaging by increasing the level of matrix metalloproteinases (MMPs), accelerating collagen degradation. Commercial dermal fillers offer temporary wrinkle reduction via volume enhancement. In this study, we propose tilapia-derived collagen hydrogels embedded with ceria nanoparticles (Ce@Col gels) as long-lasting dermal fillers for UVB-induced photoaging. Ceria nanoparticles (CeNPs) significantly enhance the stability of the collagen matrix against enzymatic degradation. These gels exhibit mechanical stability and injectability comparable to those of commercial alternatives. Additionally, CeNPs effectively eliminate ROS to suppress MMP production, curbing both collagen degradation and inflammatory responses. In a UVB-induced photoaging mouse model, the Ce@Col gels significantly reduced the level of oxidative stress in the skin, decreased the number of wrinkles, reduced epidermal thickness, and decreased levels of aging-related biomarkers while increasing the level of collagen deposition. These antiaging effects persisted for seven months post-injection, highlighting Ce@Col gels as a promising approach for prolonged collagen regeneration and sustained anti-inflammatory benefits in photoaged skin.

Indoleamine 2, 3-dioxygenase 1 inhibition mediates the therapeutic effects in Parkinson's disease mice by modulating inflammation and neurogenesis in a gut microbiota dependent manner.

Abnormal tryptophan metabolism is closely linked with neurological disorders. Research has shown that indoleamine 2,3-dioxygenase 1 (IDO-1), the first rate-limiting enzyme in tryptophan degradation, is upregulated in Parkinson's disease (PD). However, the precise role of IDO-1 in PD pathogenesis remains elusive. In this study, we administered 1-methyl-tryptophan (1-MT), an IDO-1 inhibitor, intraperitoneally at 15mg/kg daily for 21days to PD mice induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) at 30mg/kg daily for 5days. Our results show that IDO-1 inhibition improves behavioral performance, reduces dopaminergic neuron loss, and decreases serum quinolinic acid (QA) content and the aryl hydrocarbon receptor (AHR) expression in the striatum and colon. It also alleviates glial-associated neuroinflammation and mitigates colonic inflammation (decreasing iNOS, COX2) by suppressing the Toll-like receptor 4/nuclear factor-κB (TLR4/NF-κB) pathway. Furthermore, IDO-1 inhibition promotes hippocampal neurogenesis (increasing doublecortin positive (DCX+) cells and SOX2+ cells), which have recently been recognized as key pathological features and potential therapeutic targets in PD, likely through the activation of the BDNF/TrkB pathway. We further explored the gut-brain connection by depleting the gut microbiota in mice using antibiotics. Notably, the neuroprotective effects of IDO-1 inhibition were completely abolished in pseudo-germ-free mice (administrated an antibiotic mixture orally for 14days prior to 1-MT treatment), highlighting the dependency of 1-MT's neuroprotective effects on the presence of gut microbiota. Finally, we found IDO-1 inhibition corrects the abnormal elevation of fecal short chain fatty acids (SCFAs). Collectively, these findings suggest that IDO-1 inhibition may represent a promising therapeutic approach for PD.

Ghrelin recruits the endocannabinoid system to modulate food reward.

Ghrelin enhances feeding by activating the growth hormone secretagogue receptor (GHSR). In the brain, GHSRs are expressed in regions responsible for regulating food motivation including the ventral tegmental area (VTA). Endogenous cannabinoids also promote food seeking behaviors through the cannabinoid receptor 1 type (CB-1Rs) in brain regions including the VTA. It is not known, however, if ghrelin and endocannabinoids interact in the VTA to produce these effects. We therefore examined if GHSR and CB-1R interact within the VTA to enhance food motivation. Results show that GHSR and CB-1R mRNA are expressed in the VTA cells in male and female rats and mice, with the GHSR being expressed in dopamine cells, and the CB-1R being expressed primarily in non-dopaminergic cells with no obvious sex differences. Ghrelin directly activated and increased excitatory tone onto dopamine cells of male and female mice. Male rats lacking fully functional GHSR signalling showed disrupted gene expression of transcripts important for regulating the synthesis, release, and degradation of endocannabinoids and lowered the levels of 2-arachidonoylglycerol (2-AG) within the VTA. Moreover, pharmacological antagonism of VTA CB-1Rs attenuates the orexigenic and appetitive effects of intra-VTA ghrelin in rats and blocks the ability of ghrelin to promote excitatory drive to VTA dopamine neurons. Finally, blocking the breakdown of cannabinoids in the VTA enhances the effects of ghrelin on food motivation. Together, our data show that ghrelin stimulates VTA dopamine cells and ultimately food motivation in part through a mechanism that involves endocannabinoid signalling at the CB-1R.Significance Statement The current paper utilized a molecular, cellular and behavioral set of approaches to explore the interaction between the ghrelin and endocannabinoid systems in dopamine cells in the VTA. Our results not only confirm that ghrelin activates VTA dopamine neurons, and increases food motivation in rodents, but also that this interaction is in part mediated by the activation of the endocannabinoid system. We demonstrate that blocking the CB-1R in the VTA attenuates the effects of ghrelin on reward seeking behaviors and excitatory tone onto VTA dopamine neurons. Importantly, reducing enzymatic degradation of endocannabinoids enhances the effects of a sub-threshold dose of ghrelin. Overall, these data support the hypothesis that ghrelin interacts with endogenous cannabinoids to stimulate dopamine cells in the VTA.

Synthesis of a Gemcitabine Prodrug and its Encapsulation into Polymeric Nanoparticles for Improved Therapeutic Efficacy.

Gemcitabine (GEM), a chemotherapeutic agent, is widely utilized in treating various neoplasm conditions, such as pancreatic, lung, breast, and ovarian cancers. However, its therapeutic effectiveness is often hindered by its hydrophilic nature, short half-life and susceptibility to enzymatic degradation. To address these limitations, in this research, five new prodrugs of GEM were synthesized by conjugating its N-4 amino group with five different acids [4-decenoic acid (4Dec), lipoic acid (Lipo), lauric acid (Laur), 5-benzyl N-(tert-butoxycarbonyl)- L-glutamate (Glu), and decanoic acid (Dec)]. The anticancer potential of these prodrugs was evaluated using CCK-8, annexin-V FITC/propidium iodide staining, ROS, and mitochondrial membrane potential loss assays. Among the conjugates, 4Dec-GEM demonstrated comparable cytotoxic activity to native GEM. To further enhance its therapeutic efficacy, 4Dec-GEM was encapsulated in poly(lactic-co-glycolic acid) (PLGA) nanoparticles using single-emulsion and high-pressure homogenization, capitalizing on the lipophilicity of the prodrug. The developed nanoparticles were characterized by various techniques and demonstrated successful entrapment of 4Dec-GEM inside PLGA nanoparticles. Finally, the cytotoxicity of developed nanoparticles demonstrated improved therapeutic efficacy as compared to native GEM in A549, MIA-PaCa-2 and PANC-1 cancerous cell lines. Our study demonstrated that the combination of prodrug and nanoparticle can be a promising approach to augment the therapeutic efficacy of GEM.

Co-encapsulation of Creatininase, Creatinase, and Sarcosine Oxidase in Yeast Spore for Creatinine Degradation.

Creatinine clearance is used to reflect the glomerular filtration rate to assess kidney function. Creatinine degradation-related enzymes have been used for creatinine detection in clinical medicine. The mixture of spores encapsulating either creatininase or creatinase or sarcosine oxidase could mediate a three-step reaction to produce hydrogen peroxide from creatinine. In this study, to achieve consecutive and efficient creatinine detection, degradation enzymes creatininase, creatinase, and sarcosine oxidase were co-encapsulated in a single Saccharomyces cerevisiae spore. The co-encapsulation spores performed high specific activity and enzymatic properties and converted creatinine to H2O2, which was 160% higher than the mixture of spores that individually expressed these three enzymes. The detection condition of co-encapsulation was optimized for the store at room temperature and resistance to environmental stresses. The S. cerevisiae spores can co-encapsulate enzyme families and catalyze consecutive reactions in the spore wall, having potential application prospects.

The Effect of Accessibility of Insoluble Substrate on the Overall Kinetics of Enzymatic Degradation

ABSTRACTThe enzymatic reaction kinetics on cellulose and other solid substrates is limited by the access of the enzyme to the reactive substrate sites. We introduce a general model in which the reaction rate is determined by the active surface area, and the resulting kinetics consequently reflects the evolving relationship between the exposed substrate surface and the remaining substrate volume. Two factors influencing the overall surface‐to‐volume ratio are considered: the shape of the substrate particles, characterized by a single numerical parameter related to its dimensionality, and the distribution of the particle sizes. The model is formulated in a form of simple analytical equations, enabling fast and efficient application to experimental data, and facilitating its incorporation into more detailed and complex models. The application of the introduced formalism exploring its potential to account for the observed reaction rate is demonstrated on two examples: the derivation of particle size distribution from experimentally determined reaction kinetics, and the prediction of reaction slowdown from experimental particle size distribution.

Uricase from Alkalihalobacillus clausii, a Candidate for Industrial Application of Reducing Uric Acid Content of Bean Products.

Microbial uricase is an essential enzyme in purine degradation and the development of low-purine food. High enzyme activity and an appropriate optimum pH must be established for low-purine food. Uricases from Neurospora crassa, Streptomyces coelicolor, Alkalihalobacillus clausii, Bacillus subtilis, and Brevibacterium casei were heterologously expressed in Escherichia coli. Uricase from Alkalihalobacillus clausii (AC-PUCL) showed the most potent enzyme activity (249.19 IU/mL) at 37 °C and pH 7.0. This is close to the pH of plant-based food. The Km and Km/Kcat values of AC-PUCL were 30.12 μM and 1.46 s-1 μM-1, respectively. Furthermore, the crystal structures of uricases from different sources revealed that hydrogen bonds could enhance substrate affinity and strong enzyme activity. In addition, the high enzyme activity may be contributed by the active pockets with an appropriate size. Finally, AC-PUCL helped reduce the purine substances in soybean, pea, and kidney bean, with residual uric acid can not be detected at pH 8.6, suggesting a promising industrial application.

PEGylated solid lipid nanoparticles for the intranasal delivery of combination antiretroviral therapy composed of Atazanavir and Elvitegravir to treat NeuroAIDS.

Intranasal drug administration offers a promising strategy for delivering combination antiretroviral therapy (cART) directly to the central nervous system to treat NeuroAIDS, leveraging the nose-to-brain route to bypass the blood-brain barrier. However, challenges such as enzymatic degradation in the nasal mucosa, low permeability, and mucociliary clearance within the nasal cavity must first be addressed to make this route feasible. To overcome these barriers, this study developed solid lipid nanoparticles (SLNs) with varying PEGylation levels (0%, 5%, 10%, and 15% w/w of PEGylated lipid), co-encapsulated with Elvitegravir (EVG) and Atazanavir (ATZ) as an integrase and protease inhibitor, respectively. Pre-formulation studies confirmed the compatibility of the drugs with the excipients. Characterization showed that PEGylation reduces SLN size by approximately up to 12% while maintaining monodispersity and a high encapsulation efficiency of over 99% for both EVG and ATZ in their amorphous forms. Incubation of the formulations in artificial nasal mucus revealed that increased PEGylation consistently reduces nanoparticle aggregation and mean aggregate size, suggesting improved SLN stability in the mucus. Importantly, higher PEGylation levels significantly enhanced model drug permeability across the nasal mucus barrier by up to 10-fold. Lastly, cellular uptake studies using the RPMI 2650 nasal epithelial cell line indicated that PEGylation does not reduce nanoparticle uptake rates. These findings highlight the potential of PEGylated SLNs as an effective vehicle for enhancing the intranasal delivery of cART to treat NeuroAIDS. However, further in vivo studies are needed to confirm the brain targeting potential of this formulation.

Keratin nanoparticles derived from feather waste for novel antibacterial delivery.

The global rise of bacterial resistance demands innovative strategies to enhance antibiotic efficacy. This study investigates keratin nanoparticles (KNPs) derived from waste chicken feathers as sustainable drug carriers. Antibacterial activity of KNPs was evaluated against Staphylococcus aureus and Escherichia coli using antibacterial sensitivity assays, including disc diffusion and minimum inhibitory concentration tests, while cytotoxicity was evaluated on human lymphoma cells. KNPs exhibited excellent biocompatibility, showing no cytotoxic effects on human cells or bacteria. Penicillin and vancomycin were successfully loaded onto KNPs at 4 °C, 25 °C, and 50 °C temperatures for 2 and 20-hour. Loading onto KNPs enhanced the antibacterial efficacy of penicillin and vancomycin by 4-fold and 3.8-fold, respectively, against S. aureus at 37 °C. Enhanced antibacterial efficacy was attributed to molecular interactions between keratin and penicillin, as demonstrated by molecular docking analysis. The analysis revealed that the β-lactam ring of penicillin was encapsulated within the keratin matrix, potentially shielding it from enzymatic degradation by penicillinase. This protective mechanism preserves the antibiotic's structural integrity and antibacterial activity. These findings highlight the potential of KNPs as effective drug carriers in combating resistance mechanisms. This research underscores the transformative role of sustainable biological macromolecules in modern medicine, offering a promising approach to combat antibiotic resistance.

Cyclodipeptides (CDPs), the enzymatic synthesis, and the potential functional properties to exhibit broad varieties of pharmacological properties—A comprehensive review

Cyclodipeptides (CDPs) are distinct chemical scaffolds that show a wide range of bioactivities pertinent to medicine, agriculture, chemical catalysis, and material sciences. CDPs, also known as diketopiperazines (DKPs), are tiny naturally occurring peptides that have sparked interest due to their various bioactive features and possible applications in the food, pharmaceutical, and medical sectors. CDPs are produced by the intramolecular cyclization of two amino acids and can be found in a variety of environments, such as fungi, plants, animals, bacteria, and processed foods. CDPs are highly stable because of their solid structure and durability against enzymatic degradation, making them appropriate choices for medicinal and functional food applications. CDPs are frequently seen as undesirable byproducts in processed meals, especially those that include dairy, meat, and fermented drinks; new research indicates that they may improve flavor and benefit human health. Bioactive sub-stances having anti-inflammatory, antibacterial, antioxidant, and neuroprotective qualities have been recognized as CDPs. Certain CDPs, such as cyclo(Phe-Pro), along with cyclo(Pro-Pro), have shown promise in controlling metabolic and cognitive functions in the body, while others, such as cyclo(His-Pro), have demonstrated anticancer activity by causing cancer cells to undergo apoptosis. In spite of widespread research, little is known about the precise health consequences and ideal levels of CDP intake from dietary sources. Considering this, the present review attempts to compile the most recent information on the occurrence, generation, and biological activity of CDPs. To completely comprehend CDPs’ bioactivity and their significance to human health, more research is required. Additionally, creative approaches for using these peptides for creating functional foods and preventing diseases should be investigated.

Clearing the path: Unraveling bisphenol a removal and degradation mechanisms for a cleaner future.

Bisphenol A (BPA) is a prevalent chemical found in a range of consumer goods, which has raised worries about its possible health hazards. Comprehending the breakdown pathways of BPA is essential for evaluating its environmental consequences and addressing associated concerns. This review emphasizes the significance of studying the degradation/removal of BPA, with a specific focus on both natural and artificial routes. It explores natural processes such as photolysis, hydrolysis, and biodegradation, as well as manmade methods including advanced oxidation processes (AOPs) and enzymatic degradation. Examining the decomposition of BPA helps to understand how it behaves in the environment, providing valuable information for managing risks and addressing pollution. Furthermore, comprehending degradation mechanisms aids in the creation of more secure substitutes and regulatory actions to reduce BPA exposure and safeguard human health. This review emphasizes the need of promptly addressing this environmental and public health concern through the research of BPA degradation.

Sequestration of Cr(VI) onto polyethyleneimine-derivatized cellulose and its effect on the enzymatic degradation and microbiome viability

Sequestration of Cr(VI) onto polyethyleneimine-derivatized cellulose and its effect on the enzymatic degradation and microbiome viability

Fusion of two chitinases from Vibrio harveyi to enhance enzyme stability and degradation efficiency on chitin and shrimp shell

Fusion of two chitinases from Vibrio harveyi to enhance enzyme stability and degradation efficiency on chitin and shrimp shell

Self-Assembled Nanoparticle-Forming Derivatives of Dextrin-Conjugated Polyethylenimine Containing Urethane Bonds for Enhanced Delivery of Interleukin-12 Plasmid

Background and Objective: In the present investigation, low molecular weight polyethylenimine (LMW PEI, 1.8 kDa PEI) was conjugated to dextrin via urethane units and tested to transfer plasmid encoding interleukin-12 (IL-12) plasmid. Although high molecular weight PEI (HMW PEI, 25 kDa PEI) has shown substantial transfection efficiency, its wide application has been hampered due to considerable cytotoxicity. Therefore, LMW PEI with low toxic effects was used as the core of our gene transfer construct. Methods: LMW PEI was conjugated to dextrin via urethane units to improve its biophysical characteristics as well as cytotoxic effects. The conjugates were characterized in terms of buffering capacity, plasmid DNA condensation ability, particle size, and zeta potential as well as protection against enzymatic degradation. In Vitro experiments were carried out to evaluate the ability of these LMW PEI conjugates to transfer plasmid encoding human interleukin-12 (hIL- 12) to the cells. The MTT assay was performed to measure the cell-induced toxicity of the conjugates. Results: The results of our study demonstrated that the PEI derivatives with higher amounts of amine content (i.e. higher conjugation degrees) have considerable buffering capacity and plasmid condensation ability. These conjugates could condense plasmid DNA at Carrier to Plasmid ratios (C/P) ≥2 and form polyplexes at the size range of 120-165 nm while their zeta potential was around 5.5-8.5 mV. The results of transfection efficiency demonstrated that the level of IL- 12 production increased by 2-3 folds compared with unmodified LMW PEI while the level of cytotoxicity was not higher than 20%. Conclusion: The strategy used in this study shows a promising way to prepare gene carriers with high transfection efficiency and low toxicity.

Size-controlled antimicrobial peptide drug delivery vehicles through complex coacervation.

Due to the escalating threat of the pathogens' capability of quick adaptation to antibiotics, finding new alternatives is crucial. Although antimicrobial peptides (AMPs) are highly potent and effective, their therapeutic use is limited' as they are prone to enzymatic degradation, are cytotoxic and have low retention. To overcome these challenges, we investigate the complexation of the cationic AMP colistin with diblock copolymers poly(ethylene oxide)-b-poly(methacrylic acid) (PEO-b-PMAA) forming colistin-complex coacervate core micelles (colistin-C3Ms). We present long-term stable kinetically controlled colistin-C3Ms that can be prepared from several block lengths of PEO-b-PMAA polymers, where the polymerisation degree governs the overall micellar size. To achieve precise control over size and polydispersity, which are crucial for drug delivery applications, we investigate the hybridisation of PEO-b-PMAA polymers with varying chain lengths or PMAA homopolymers in ternary complex coacervation systems with colistin. This results in size-tunable colistin-C3Ms, ranging, depending on the mixing ratios, from micellar sizes of 26 nm to 100 nm. With size tunability at rather narrow size distributions and high stability, ternary colistin-C3Ms offer potential advancements in C3M drug delivery, paving the way for more effective and targeted treatments for bacterial infections in precision medicine.