RESUMEN
This work was aimed at producing peptides containing iron-binding capabilities from scad (Decapterus maruadsi) processing by-product with alcalase hydrolysis. The chelating peptides were purified by ultrafiltration, immobilized-metal affinity chromatography, gel filtration chromatography, and reversed-phase high-performance liquid chromatography. A novel iron-binding peptide was purified with 1,386.63 Da molecular weight and amino acid sequence of QKGTYDDYVEGL. The peptide binds to iron mainly through carboxyl and hydroxyl oxygen bonds. The iron-binding peptide can significantly promote the absorption of inorganic iron in Caco-2 cells. These results have contributed to development of the peptide from scad processing by-products hydrolyzate in iron supplementations. PRACTICAL APPLICATIONS: Iron deficiency is one of the most common and widespread nutritional disorders in the world. Iron-peptide chelates may be suitable for iron-fortification. Our study shows that a peptide purified from scad processing by-product has iron-chelating activity, and significantly increases iron absorption by Caco-2 cells. Hence, this peptide has potential application as a novel carrier for enhancing iron absorption.
Asunto(s)
Quelantes del Hierro/química , Péptidos/química , Residuos/análisis , Animales , Biocatálisis , Células CACO-2 , Cromatografía de Afinidad , Cromatografía de Fase Inversa , Proteínas de Peces/química , Humanos , Hierro/química , Quelantes del Hierro/aislamiento & purificación , Péptidos/aislamiento & purificación , Subtilisinas/químicaRESUMEN
Plantaricin NC8, a two-peptide non-lantibiotic class IIb bacteriocin composed of PLNC8α and PLNC8ß and derived from Lactobacillus plantarum ZJ316, has been shown to be highly potent against a range of bacteria and fungi. In this study, we assessed the antimicrobial mechanism of plantaricin NC8 against the most sensitive bacterial strain, Micrococcus luteus CGMCC 1.193. The results showed that plantaricin NC8 induced membrane permeabilization and caused cell membrane disruption to M. luteus CGMCC 1.193 cells, as evidenced by electrolyte efflux, loss of proton motive force, and ATP depletion within a few minutes of plantaricin NC8 treatment. Furthermore, scanning and transmission electron microscopy showed that plantaricin NC8 had a drastic impact on the structure and integrity of M. luteus CGMCC 1.193 cells. In addition, we found that either PLNC8α or PLNC8ß alone exhibited membrane permeabilization activity, but that PLNC8ß had higher permeabilization activity, and their individual effects were not as strong as that of the combined compounds as plantaricin NC8. Finally, we showed that lipid II is not the specific target of plantaricin NC8 against M. luteus CGMCC 1.193. Our study reveals the antimicrobial mechanism of plantaricin NC8 against M. luteus CGMCC 1.193.
Asunto(s)
Bacteriocinas/farmacología , Lactobacillus plantarum/química , Micrococcus luteus/efectos de los fármacos , Bacteriocinas/aislamiento & purificación , Membrana Celular/efectos de los fármacos , Uridina Difosfato Ácido N-Acetilmurámico/análogos & derivados , Uridina Difosfato Ácido N-Acetilmurámico/metabolismoRESUMEN
Bacteriocins are bioactive antimicrobial peptides synthesized in the ribosome of numerous bacteria and released extracellularly. Bacteriocins have the ability to kill or inhibit the growth of prokaryotes and could potentially be useful against pathogens and antibiotic-resistant strains of bacteria. The antimicrobial mechanisms and relatively narrow killing spectrums of bacteriocins distinguish them from traditional broad-spectrum antibiotics, making them possible candidates to replace antibiotics in the future. In recent years, researchers have discovered many new bacteriocins from different sources. However, the discovery of new bacteriocins involves complicated screening, identification, purification, and characterization processes. This review describes the strategies currently employed for screening, identification, purification, and characterization of bacteriocins. Approximately 40 novel bacteriocins are reviewed in this paper. Our conclusions about the research process will guide the development of novel methods and allow for the faster, easier, and more efficient discovery of novel bacteriocins.
Asunto(s)
Bacteriocinas/análisis , Bacteriocinas/aislamiento & purificación , Fraccionamiento Químico/métodos , Animales , Bacteriocinas/química , HumanosRESUMEN
Well-defined molecular brushes bearing polypeptides as side chains were prepared by a "grafting through" synthetic strategy with two-dimensional control over the brush molecular architectures. By integrating N-carboxyanhydride ring-opening polymerizations (NCA ROPs) and ring-opening metathesis polymerizations (ROMPs), desirable segment lengths of polypeptide side chains and polynorbornene brush backbones were independently constructed in controlled manners. The N2 flow accelerated NCA ROP was utilized to prepare polypeptide macromonomers with different lengths initiated from a norbornene-based primary amine, and those macromonomers were then polymerized via ROMP. It was found that a mixture of dichloromethane and an ionic liquid were required as the solvent system to allow for construction of molecular brush polymers having densely-grafted peptide chains emanating from a polynorbornene backbone, poly(norbornene-graft-poly(ß-benzyl-l-aspartate)) (P(NB-g-PBLA)). Highly efficient postpolymerization modification was achieved by aminolysis of PBLA side chains for facile installment of functional moieties onto the molecular brushes.
RESUMEN
A polypeptide-based hydrogel system, when prepared from a diblock polymer with a ternary copolypeptide as one block, exhibited thermo-, mechano- and enzyme-responsive properties, which enabled the encapsulation of naproxen (Npx) during the sol-gel transition and its release in the gel state. Statistical terpolymerizations of l-alanine (Ala), glycine (Gly) and l-isoleucine (Ile) NCAs at a 1 : 1 : 1 feed ratio initiated by monomethoxy monoamino-terminated poly(ethylene glycol) afforded a series of methoxy poly(ethylene glycol)-block-poly(l-alanine-co-glycine-co-l-isoleucine) (mPEG-b-P(A-G-I)) block polymers. ß-Sheets were the dominant secondary structures within the polypeptide segments, which facilitated a heat-induced sol-to-gel transition, resulting from the supramolecular assembly of ß-sheets into nanofibrils. Deconstruction of the three-dimensional networks by mechanical force (sonication) triggered the reverse gel-to-sol transition. Certain enzymes could accelerate the breakdown of the hydrogel, as determined by in vitro gel weight loss profiles. The hydrogels were able to encapsulate and release Npx over 6 days, demonstrating the potential application of these polypeptide hydrogels as an injectable local delivery system for small molecule drugs.
Asunto(s)
Anhídridos/química , Antiinflamatorios no Esteroideos/química , Sistemas de Liberación de Medicamentos , Hidrogeles/química , Péptidos/química , Hidrogeles/síntesis química , Estructura Molecular , Péptidos/síntesis química , PolimerizacionRESUMEN
Staphylococcus aureus and its drug-resistant strains, which threaten public health and food safety, are in need of effective control by biopreservatives. A novel bacteriocin, pentocin JL-1, produced by Lactobacillus pentosus that was isolated from the intestinal tract of Chiloscyllium punctatum, was purified by a four-step chromatographic process. Mass spectrometry based on MALDI-TOF indicated that pentocin JL-1 has a molecular mass of 2987.23 Da. Only six of the twenty-five amino acids could be identified by Edman degradation. This bacteriocin is thermostable and tolerates a pH range of 5-7. Also, it is sensitive to proteinase K, trypsin, pepsin, and alkaline protease. This bacteriocin has a broad inhibitory spectrum against both Gram-positive and Gram-negative strains and in particular is effective against multidrug-resistant S. aureus. Additionally, we showed that the cell membrane is the target of pentocin JL-1 against methicillin-resistant S. aureus (MRSA), causing a loss of proton motive force. Furthermore, pentocin JL-1 has a drastic impact on the structure and integrity of MRSA cells. These results suggest that pentocin JL-1 has potential as a biopreservative in the food industry.
Asunto(s)
Antibacterianos/farmacología , Bacteriocinas/química , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Infecciones Estafilocócicas/tratamiento farmacológico , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Bacteriocinas/aislamiento & purificación , Bacteriocinas/farmacología , Humanos , Lactobacillus pentosus/química , Espectrometría de Masas , Staphylococcus aureus Resistente a Meticilina/patogenicidad , Peso Molecular , Infecciones Estafilocócicas/microbiologíaRESUMEN
A strategy for reversible patterning of soft conductive materials is described, based upon a combination of peptide-based block copolymer hydrogelators and photo-thermally-active carbon nanotubes. This composite displays photo-responsive gelation at application-relevant timescales (<10 s), allowing for rapid and spatially-defined construction of conductive patterns (>100 S m(-1)), which, additionally, hold the capability to revert to sol upon sonication for reprocessing.
RESUMEN
Using the 3(rd) generation Grubbs' catalyst as the initiator, diblock brush polymer drug conjugates (BPDCs) were synthesized by sequential ring-opening metathesis polymerization (ROMP) of a hydrophilic poly(ethylene glycol) (PEG)-based norbornene (NB)-functionalized macromonomer and a hydrophobic paclitaxel (PTXL)-based NB-functionalized monomer. These amphiphilic diblock BPDCs had well-defined structures, with narrow molecular weight distributions (Mw/Mn = 1.10-1.16). They self-assembled into multi-molecular nanostructures in aqueous solutions. Although the PTXL moieties were connected to the backbone with cycloacetal-based conjugation linkages, the cleavage of these linkages from the assemblies of diblock BPDCs was relatively slow and exhibited limited acid-sensitivity, indicating a significant influence of the macromolecular structure and assembly of BPDCs on their drug release behaviour. The cytotoxicity study not only showed that the diblock BPDCs are therapeutically effective against cancer cells, but also revealed a correlation between cytotoxicity and grafting structures of BPDCs. In summary, the results obtained in this work provide new insight into the structure-dependent properties of brush polymer-based drug delivery systems.
Asunto(s)
Portadores de Fármacos/química , Sistemas de Liberación de Medicamentos/métodos , Sustancias Macromoleculares/química , Norbornanos/química , Paclitaxel/química , Paclitaxel/farmacocinética , Polímeros/química , Caproatos/química , Línea Celular , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Lactonas/química , Estructura Molecular , Paclitaxel/administración & dosificación , Polietilenglicoles/química , Polimerizacion , Sales de Tetrazolio/química , Tiazoles/químicaRESUMEN
Nanomaterials have great potential to offer effective treatment against devastating diseases by providing sustained release of high concentrations of therapeutic agents locally, especially when the route of administration allows for direct access to the diseased tissues. Biodegradable polyphosphoester-based polymeric micelles and shell cross-linked knedel-like nanoparticles (SCKs) have been designed from amphiphilic block-graft terpolymers, PEBP-b-PBYP-g-PEG, which effectively incorporate high concentrations of paclitaxel (PTX). Well-dispersed nanoparticles physically loaded with PTX were prepared, exhibiting desirable physiochemical characteristics. Encapsulation of 10 wt% PTX, into either micelles or SCKs, allowed for aqueous suspension of PTX at concentrations up to 4.8 mg/mL, as compared to <2.0 µg/mL for the aqueous solubility of the drug alone. Drug release studies indicated that PTX released from these nanostructures was defined through a structure-function relationship, whereby the half-life of sustained PTX release was doubled through cross-linking of the micellar structure to form SCKs. In vitro, physically loaded micellar and SCK nanotherapeutics demonstrated IC50 values against osteosarcoma cell lines, known to metastasize to the lungs (CCH-OS-O and SJSA), similar to the pharmaceutical Taxol formulation. Evaluation of these materials in vivo has provided an understanding of the effects of nanoparticle structure-function relationships on intratracheal delivery and related biodistribution and pharmacokinetics. Overall, we have demonstrated the potential of these novel nanotherapeutics toward future sustained release treatments via administration directly to the sites of lung metastases of osteosarcoma.
Asunto(s)
Portadores de Fármacos/química , Nanopartículas/química , Paclitaxel/química , Polietilenglicoles/química , Polímeros/química , Alquinos/química , Animales , Azidas/química , Neoplasias Óseas/patología , Catálisis , Línea Celular Tumoral , Cobre/química , Portadores de Fármacos/metabolismo , Portadores de Fármacos/farmacocinética , Liberación de Fármacos , Ésteres , Semivida , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundario , Ratones , Micelas , Modelos Moleculares , Conformación Molecular , Osteosarcoma/patología , Polímeros/metabolismo , Polímeros/farmacocinética , Distribución TisularRESUMEN
An antibiofouling polymer coating, combined with both zwitterionic and amphiphilic features, is engineered by a two-step modification of a commodity polymer. The surface properties of the resultant polymer coating can be easily tuned by varying the extent of cross-linking in the network. Higher antibiofouling efficiency was observed for these surfaces vs. an elastomeric polydimethylsiloxane standard (Sylgard 184) against the adsorption of biomacromolecules and a marine fouling organism (Ulva zoospores) has been demonstrated. This design establishes a platform for the achievement of functionalized amphiphilic zwitterionic copolymers from relatively inexpensive starting materials via simple chemical manipulations.
RESUMEN
A multi-responsive triblock hydrogelator oligo(dl-allylglycine)-block-poly(ethylene glycol)-block-oligo(dl-allylglycine) (ODLAG-b-PEG-b-ODLAG) was synthesized facilely by ring-opening polymerization (ROP) of DLAG N-carboxyanhydride (NCA) with a diamino-terminated PEG as the macroinitiator. This system exhibited heat-induced sol-to-gel transitions and either sonication- or enzyme-induced gel-to-sol transitions. The ß-sheeting of the oligopeptide segments was confirmed by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and wide-angle X-ray scattering (WAXS). The ß-sheets further displayed tertiary ordering into fibrillar structures that, in turn generated a porous and interconnected hydrogel matrix, as observed via transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The reversible macroscopic sol-to-gel transitions triggered by heat and gel-to-sol transitions triggered by sonication were correlated with the transformation of nanostructural morphologies, with fibrillar structures observed in gel and spherical aggregates in sol, respectively. The enzymatic breakdown of the hydrogels was also investigated. This allyl-functionalized hydrogelator can serve as a platform for the design of smart hydrogels, appropriate for expansion into biological systems as bio-functional and bio-responsive materials.
RESUMEN
Sequential polymerization of N-carboxyanhydrides accelerated by nitrogen flow is utilized to generate a novel well-defined diblock copolypeptide (PDI = 1.08), with incorporation of alkyne-functionalized side-chain groups allowing for rapid and efficient thiol-yne click-type modifications, followed by self-assembly into nanopure water to construct a helical polypeptide-based versatile and functional nanoparticle platform.
RESUMEN
A facile polymerization of an allyl-functionalized N-carboxyanhydride (NCA) monomer is utilized to construct an A-B-A-type triblock structure containing ß-sheet-rich oligomeric peptide segments tethered by a poly(ethylene oxide) chain, which are capable of dispersing and gelating single-walled carbon nanotubes (SWCNTs) noncovalently in organic solvents, resulting in significant enhancement of the mechanical properties of polypeptide-based organogels.
Asunto(s)
Geles/química , Nanotubos de Carbono/química , Oligopéptidos/química , Nanotubos de Carbono/ultraestructura , Polietilenglicoles/química , Polimerizacion , Estructura Secundaria de ProteínaRESUMEN
The simple copolymerization of N-carboxyanhydride (NCA) monomers is utilized to generate copolypeptides having a combination of α-helix and ß-sheet sub-structures that, when grown from a solvophilic synthetic polymer block segment, are capable of driving mechano-responsive supramolecular sol-to-gel-to-sol and sol-to-gel-to-gel transitions reversibly, which allow also for injection-based processing and self-healing behaviors. A new type of polypeptide-based organogelator, methoxy poly(ethylene glycol)-block-poly(γ-benzyl-l-glutamate-co-glycine) (mPEG-b-P(BLG-co-Gly)), is facilely synthesized by statistical ring-opening copolymerizations (ROPs) of γ-benzyl-l-glutamate (BLG) and glycine (Gly) NCAs initiated by mPEG-amine. These systems exhibit tunable secondary structures and result in sonication stimulus responsiveness of the organogels with the polypeptide segment variation, controlled by varying the ratio of BLG NCA to Gly NCA during the copolymerizations. Attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) studies indicate the α-helical component decreases while the ß-sheet content increases systematically with a higher mole fraction of Gly in the polypeptide segment. The supramolecular assembly of ß-sheet nanofibrils, having a tunable width over the range of 10.4 - 14.5 nm with varied BLG to Gly ratio, are characterized by transmission electron microscopy (TEM). The further self-assembly of these nanostructures into 3-D gel networks within N,N-dimethylformamide (DMF) occurs at low critical gelation concentrations (CGC) (lowest ca. 0.6 wt %). Increased BLG to Gly ratios lead to an increase of the α-helical component in the secondary structures of the polypeptide segments, resulting in wider and more flexible nanofibrils. The presence of α-helical component in the polymers enhances the stability of the organogels against sonication, and instantaneous gel-to-gel transitions are observed as in situ reconstruction of networks occurs within the gelled materials after sonication. In marked contrast, the ß-sheet-rich gel, prepared from mPEG-b-PGly, exhibits an instant gel-to-sol transition after sonication is applied. The CGC concentration and stiffness of this mPEG-b-P(BLG-co-Gly) organogel system can be tuned by simply varying the percentages of α-helix and ß-sheet in the secondary structures through control of the BLG to Gly ratio during synthesis. The mechanical properties of these organogels are studied by dynamic mechanical analyses (DMA), having storage moduli of ca. 12.1 kPa at room temperature. The injectability and self-healing capabilities are demonstrated by direct observation of the macroscopic self-healing behavior experiment.
RESUMEN
Electrostatic interaction-mediated enzymatic-hydrolysis of poly(lactide)-containing nanoscale assemblies is described. At physiological pH, degradable core-shell morphologies with charged shells can readily attract or repel enzymes carrying opposite or similar charges, respectively.
Asunto(s)
Enzimas/metabolismo , Poliésteres/metabolismo , Enzimas/química , Concentración de Iones de Hidrógeno , Hidrólisis , Micelas , Nanoestructuras/química , Poliésteres/química , Electricidad EstáticaRESUMEN
There has been an increasing interest to develop new types of stimuli-responsive drug delivery vehicles with high drug loading and controlled release properties for chemotherapeutics. An acid-labile poly(ethylene oxide)-block-polyphosphoester-graft-PTX drug conjugate (PEO-b-PPE-g-PTX G2) degradable, polymeric paclitaxel (PTX) conjugate containing ultra-high levels of PTX loading is improved significantly, in this second-generation development, which involves connection of each PTX molecule to the polymer backbone via a pH-sensitive ß-thiopropionate linkage. The PEO-b-PPE-g-PTX G2 forms well-defined nanoparticles in an aqueous solution, by direct dissolution into water, with a number-averaged hydrodynamic diameter of 114 ± 31 nm, and exhibits a PTX loading capacity as high as 53 wt%, with a maximum PTX concentration of 0.68 mg mL(-1) in water (vs 1.7 µg mL(-1) for free PTX). The PEO-b-PPE-g-PTX G2 shows accelerated drug release under acidic conditions (≈50 wt% PTX released in 8 d) compared with neutral conditions (≈20 wt% PTX released in 8 d). Compared to previously reported polyphosphoester-based PTX drug conjugates, PEO-b-PPE-g-PTX G1 without the ß-thiopropionate linker, the PEO-b-PPE-g-PTX G2 shows pH-triggered drug release property and 5- to 8-fold enhanced in vitro cytotoxicity against two cancer cell lines.
Asunto(s)
Antineoplásicos/química , Organofosfatos/síntesis química , Paclitaxel/análogos & derivados , Paclitaxel/química , Polietilenglicoles/química , Animales , Antineoplásicos/farmacología , Línea Celular Tumoral , Sistemas de Liberación de Medicamentos/métodos , Concentración de Iones de Hidrógeno , Concentración 50 Inhibidora , Espectroscopía de Resonancia Magnética , Ratones , Nanopartículas/administración & dosificación , Nanopartículas/química , Organofosfatos/farmacología , Paclitaxel/síntesis química , Paclitaxel/farmacología , Tamaño de la Partícula , Polietilenglicoles/síntesis química , Polietilenglicoles/farmacología , Polímeros/químicaRESUMEN
The direct synthesis of an acid-labile polyphosphoramidate by organobase-catalyzed ring-opening polymerization and an overall two-step preparation of polyphosphodiester ionomers (PPEI) by acid-assisted cleavage of the phosphoramidate bonds along the backbone of the polyphosphoramidate were developed in this study. The ultrafast organobase-catalyzed ring-opening polymerization of a cyclic phospholane methoxyethyl amidate monomer initiated by benzyl alcohol allowed for the preparation of well-defined polyphosphoramidates (PPA) with predictable molecular weights, narrow molecular weight distributions (PDI<1.10), and well-defined chain ends. Cleavage of the acid-labile phosphoramidate bonds on the polyphosphoramidate repeat units was evaluated under acidic conditions over a pH range of 1-5, and the complete hydrolysis produced polyphosphodiesters. The thermal properties of the resulting polyphosphoester ionomer acid and polyphosphoester ionomer sodium salt exhibited significant thermal stability. The parent PPA and both forms of the PPEIs showed low cytotoxicities toward HeLa cells and RAW 264.7 mouse macrophage cells. The synthetic methodology developed here has enriched the family of water-soluble polymers prepared by rapid and convenient organobase-catalyzed ring-opening polymerizations and straightforward chemical medication reactions, which are designed to be hydrolytically degradable and have promise for numerous biomedical and other applications.
RESUMEN
A degradable polyphosphoester (PPE)-based cationic nanoparticle (cSCK), which is integrated constructed as a novel degradable drug device, demonstrates surprisingly efficient inhibition of inducible nitric oxide synthase (iNOS) transcription, and eventually inhibits nitric oxide (NO) over-production, without loading of any specific therapeutic drugs. This system may serve as a promising anti-inflammatory agent toward the treatment of acute lung injury.
Asunto(s)
Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/farmacología , Nanopartículas/química , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Polímeros/química , Polímeros/farmacología , Animales , Transporte Biológico , Línea Celular , Inhibidores Enzimáticos/metabolismo , Ésteres , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Óxido Nítrico/biosíntesis , Polímeros/metabolismoRESUMEN
We demonstrate the improved second-harmonic Talbot self-imaging through the quasi-phase-matching technique in a 2D periodically-poled LiTaO(3) crystal. The domain structure not only composes a nonlinear optical grating which is necessary to realize nonlinear Talbot self-imaging, but also provides reciprocal vectors to satisfy the phase-matching condition for second-harmonic generation. Our experimental results show that quasi-phase-matching can improve the intensity of the second-harmonic Talbot self-imaging by a factor of 21.