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1.
Food Chem ; 462: 140995, 2025 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-39213970

RESUMEN

The storage and processing of Litopenaeus vannamei are often challenged by the freeze-thaw (F-T) cycle phenomenon. This study delved into the influence of pretreatment with l-arginine (Arg) and l-lysine (Lys) on the myofibrillar proteins oxidation and quality of shrimp subjected to F-T cycles. Arg and Lys pretreatment notably improved water-holding capacity (WHC), textural integrity as well as the myofibrillar structure of the shrimps. A lesser reduction in the amounts of immobile and bound water was found in the amino acid-treated groups, and the oxidation of lipids and proteins were both decelerated. Molecular simulation results indicated that Arg and Lys could form hydrogen and salt-bridge bonds with myosin, enhancing the stability of Litopenaeus vannamei. The study concludes that Arg and Lys are effective in alleviating the adverse effects of F-T cycles on the quality of Litopenaeus vannamei, and provides a new solution for the quality maintenance during storage and processing.


Asunto(s)
Arginina , Lisina , Proteínas Musculares , Oxidación-Reducción , Penaeidae , Animales , Penaeidae/química , Arginina/química , Lisina/química , Proteínas Musculares/química , Congelación , Conservación de Alimentos/métodos , Mariscos/análisis , Miofibrillas/química
2.
Artículo en Inglés | MEDLINE | ID: mdl-39262294

RESUMEN

OBJECTIVES: This study aimed to explore multiple effects of hyperbranched poly-l-lysine (HBPL) titanium (Ti) surfaces on osseointegration, bacteriostasis, and anti-inflammation across three different animal models. METHODS: Ti surfaces were covalently modified with HBPL, with uncoated surfaces as controls. Characterization included scanning electron microscopy (SEM) and surface chemistry and elemental analysis (EDX). Ti and Ti-HBPL implants were placed in conventional canine edentulous sites, post-operative infection canine edentulous sites, and diabetic rat tibias. Implants from canine edentulous models were analyzed using micro-CT and histomorphometry to assess osseointegration at 8 weeks. Post-operative infection beagles were used to evaluate antibacterial efficacy through clinical parameters and bacterial cultures at 1 week. In diabetic rats, micro-CT and histomorphometry were performed at 8 weeks. RESULTS: HBPL was uniformly grafted on Ti-HBPL surfaces. Ti-HBPL surfaces showed higher bone volume/total volume (BV/TV, p < 0.001), bone-implant contact (BIC%, p < 0.001), and trabecular number (Tb.N, p < 0.01) in beagles. Besides, it displayed higher BIC% (p < 0.001) and bone area fraction occupancy (BAFO%, p < 0.01) in hard tissue sections. In an infected model, Ti-HBPL surfaces exhibited lower bleeding on probing (BOP, p < 0.001), and plaque index (DI, p < 0.01), with reduced bacterial colony formation (p < 0.001) compared to the control group. In diabetic rats, Ti-HBPL surfaces showed an increase in BV/TV (p < 0.01) and Tb.N (p < 0.001), downregulated TNF-α and IL-1ß (p < 0.01), and upregulated IL-10 (p < 0.01) and osteocalcin (OCN) expression (p < 0.01). CONCLUSIONS: HBPL-Ti surfaces demonstrated enhanced osseointegration, bacteriostasis, and anti-inflammatory effects in vivo.

3.
J Chromatogr A ; 1733: 465228, 2024 Sep 27.
Artículo en Inglés | MEDLINE | ID: mdl-39163701

RESUMEN

In this study, using chiral L-lysine as the molecular skeleton, three kinds of L-lysine-derived gelators (GBLB, GBLF and GFLF) were synthesized and then bonded to the surface of silica matrix (5 µm) by amide condensation to prepare a series of multifunctional chromatography stationary phases (GBLB-SiO2, GBLF-SiO2, and GFLF-SiO2) were prepared. The L-lysine-derived gelators not only possess chiral recognition ability, but also can spontaneously form oriented and ordered network structures in liquid medium through the interaction of non-covalent bonding forces such as hydrogen bonding, π-π stacking, and van der Waals forces. The comprehensive effect of multiple weak interaction sites enhances the molecular recognition ability and further improves the separation diversity of different types of compounds on stationary phases. The separation and evaluation of chiral compounds showed that benzoin, 1-phenyl-ethanol, 1-phenyl-propanol and 6-hydroxyflavanone could be separated in normal phase mode (NPLC). The separation of different types of non-chiral compounds, such as sulfonamides, nucleosides, nucleobases, polycyclic aromatic hydrocarbons (PAHs), anilines, and aromatic acids, were achieved in hydrophilic interaction/reversed-phase/ion-exchange mode (HILIC/RPLC/IEC), and the separation of polarized compounds could be performed under the condition of ultrapure water as the mobile phase, which has the typical retention characteristics of per aqueous liquid chromatography (PALC). The effects of organic solvent content, temperature, pH value, and buffer salt concentration on the retention and separation performance of the column were investigated. Comparison of the three prepared columns showed that the separation performance (such as aromatic selectivity) could be improved by increasing the types of functional groups on the surface of the stationary phase and the number of aromatic groups. In a word, the prepared stationary phase have multiple retention properties, can simultaneously separate chiral compounds and various types of achiral compounds. This work provides an idea for developing multifunctional liquid chromatography stationary phase materials, and further expands the application of gelators in separation science.


Asunto(s)
Interacciones Hidrofóbicas e Hidrofílicas , Lisina , Dióxido de Silicio , Estereoisomerismo , Lisina/química , Dióxido de Silicio/química , Cromatografía Líquida de Alta Presión/métodos
4.
ACS Synth Biol ; 13(9): 2948-2959, 2024 Sep 20.
Artículo en Inglés | MEDLINE | ID: mdl-39158285

RESUMEN

5-Aminovalerate (5-AVA), 5-hydroxyvalerate (5-HV), and 1,5-pentanediol (1,5-PDO) are l-lysine derivatives with extensive applications in the production of materials such as polyesters, polyurethane, plasticizers, inks, and coatings. However, their large-scale production is limited by the lack of efficient synthetic pathways. Here, we aimed to construct multiple synthetic pathways by screening the key enzymes involved in the synthesis of these compounds in Escherichia coli. The engineered pathway utilizing RaiP demonstrated a superior catalytic efficiency. The LER strain that overexpressed only raiP successfully synthesized 9.70 g/L 5-HV and 8.31 g/L 5-AVA, whereas the strain LERGY that overexpressed raiP, gabT, and yahK accumulated 9.72 g/L 5-HV and 7.95 g/L 5-AVA from 20 g/L glucose. The introduction of exogenous transaminases and dehydrogenases enhanced cell growth and fermentation efficiency with respect to 5-HV synthesis, albeit without significantly impacting the yield. Strain LE05, incorporating only two exogenous enzymes, RaiP and CaR, produced 1.87 g/L 1,5-PDO, 3.85 g/L 5-HV, and 4.78 g/L 5-hydroxyglutaraldehyde from 20 g/L glucose after 6 days. The strain LE02G, fortified with transaminase, dehydrogenase, and NADPH regeneration system, accumulated 7.82 g/L 1,5-PDO, whereas the aldp-knock out LE02G2 synthesized 10.98 g/L 1,5-PDO from 50 g/L glucose in fed-batch fermentation after 6 days, yielding 0.22 g/g glucose (0.37 mol/mol). Introducing the NADPH regeneration pathway and deleting the NADPH-consuming pathways increased the 1,5-PDO yield and decreased the precursor concentration. The proposed pathways and engineering strategies presented in this study can prove instrumental in developing biological routes for the practical production of 5-AVA, 5-HV, and 1,5-PDO.


Asunto(s)
Escherichia coli , Lisina , Ingeniería Metabólica , Ingeniería Metabólica/métodos , Escherichia coli/genética , Escherichia coli/metabolismo , Lisina/metabolismo , Fermentación , Glucosa/metabolismo , Transaminasas/metabolismo , Transaminasas/genética , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Aminoácidos Neutros
5.
Nutrients ; 16(16)2024 Aug 14.
Artículo en Inglés | MEDLINE | ID: mdl-39203831

RESUMEN

Various nutritional supplements are available over the counter, yet few have been investigated in randomized controlled trials. The rationale for using the specific mix of nutritional substances including collagen type II, hyaluronic acid, n-acetyl-glucosamine, bamboo extract, L-lysine, and vitamin C is the assumption that combining naturally occurring ingredients of the intervertebral disc would maintain spine function. This double-blinded, placebo-controlled randomized trial aimed to evaluate the efficacy of a nutraceutical supplement mix in the management of lumbar osteochondrosis. Fifty patients were randomly assigned to either the supplement or placebo group in a 1:1 ratio. Patient-Reported Outcome Measures (PROMs) included the Oswestry Disability Index (ODI), the visual analogue scale for pain (pVAS), short form-12 (SF-12) physical and mental component summary subscale scores (PCS and MCS, respectively), and global physical activity questionnaire (GPAQ). Magnetic resonance imaging (MRI) was used to evaluate degenerative changes of intervertebral discs (IVD) including Pfirrmann grades as well as three-dimensional (3D) volume measurements. Data were collected at baseline and after the 3-month intervention. None of the PROMs were significantly different between the supplement and placebo groups. Disc degeneration according to Pfirrmann classifications remained stable during the 3-month intervention in both groups. Despite no significance regarding the distribution of Pfirrmann grade changes (improvement, no change, worsening; p = 0.259), in the supplement group, one patient achieved a three-grade improvement, and worsening of Pfirrmann grades were only detected in the placebo group (9.1%). Furthermore, in-depth evaluations of MRIs showed significantly higher 3D-measured volume changes (increase) in the supplement (+740.3 ± 796.1 mm3) compared to lower 3D-measured volume changes (decrease) in the placebo group (-417.2 ± 875.0 mm3; p < 0.001). In conclusion, this multi-nutrient supplement might not only stabilize the progression of lumbar osteochondrosis, but it might also potentially even increase IVD volumes as detected on MRIs.


Asunto(s)
Suplementos Dietéticos , Vértebras Lumbares , Humanos , Método Doble Ciego , Masculino , Femenino , Persona de Mediana Edad , Vértebras Lumbares/diagnóstico por imagen , Adulto , Degeneración del Disco Intervertebral , Resultado del Tratamiento , Imagen por Resonancia Magnética , Medición de Resultados Informados por el Paciente , Osteocondrosis/terapia , Osteocondrosis/tratamiento farmacológico , Osteocondrosis de la Columna Vertebral/terapia , Osteocondrosis de la Columna Vertebral/tratamiento farmacológico , Dimensión del Dolor
6.
Int J Biol Macromol ; 277(Pt 4): 134535, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39111467

RESUMEN

Microbial enzymes are crucial catalysts in various industries due to their versatility and efficiency. The microbial enzymes market has recently expanded due to increased demand for many reasons. Among them are eco-friendly solutions, developing novel microbial strains with enhanced enzymes that perform under harsh conditions, providing sustainability, and raising awareness about the benefits of enzyme-based products. By 2030, the global enzyme market is expected to account for $525 billion, with a growth rate of 6.7 %. L-asparaginase and L-glutaminase are among the leading applied microbial enzymes in antitumor therapy, with a growing market share of 16.5 % and 9.5 %, respectively. The use of microbial enzymes has opened new opportunities to fight various tumors, including leukemia, lymphosarcoma, and breast cancer, which has increased their demand in the pharmaceutical and medicine sectors. Despite their promising applications, commercial use of microbial enzymes faces challenges such as short half-life, immunogenicity, toxicity, and other side effects. Therefore, this review explores the industrial production, purification, formulation, and commercial utilization of microbial enzymes, along with an overview of the global enzyme market. With ongoing discoveries of novel enzymes and their applications, enzyme technology offers promising avenues for cancer treatment and other therapeutic interventions.


Asunto(s)
Neoplasias , Humanos , Neoplasias/tratamiento farmacológico , Neoplasias/enzimología , Neoplasias/terapia , Antineoplásicos/uso terapéutico , Antineoplásicos/farmacología , Antineoplásicos/química , Asparaginasa/uso terapéutico , Asparaginasa/química , Asparaginasa/metabolismo , Glutaminasa/metabolismo , Bacterias/enzimología
7.
Heliyon ; 10(15): e35264, 2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-39161815

RESUMEN

Hydrogels containing antimicrobial materials have emerged as attractive platforms for wound treatment in the past decade due to their favorable bio-mimicking properties, excellent modulation of bacterial infection, and ability to minimize bacterial resistance. Herein, a hybrid combination of polyvinyl alcohol (PVA), hyperbranched poly L-lysine (L), tannic acid decorated AgNPs (AgTA NPs), loaded with Allantoin (Alla) is used to fabricate PLAg-Alla hydrogel dressing via the freeze-thaw method without use of any chemical cross-linker. The PLAg-Alla hydrogel possesses a great structure, is biodegradable, and safe, and exhibits high antibacterial potential, all required for efficient wound healing. The incorporation of AgTA and poly L-lysine (L) within the hydrogel contributes to the enhancement of antibacterial ability, as well as effectively promoting the wound healing. This hybrid hydrogel possessed favorable physicochemical features, robust antibacterial properties, and accelerated wound healing in vivo as promising dressing for the clinical application.

8.
Artículo en Inglés | MEDLINE | ID: mdl-39150530

RESUMEN

ε-Poly-L-lysine (ε-PL) is a natural and wide-spectrum antimicrobial additive. In this study, the production of ε-PL by Streptomyces albulus FQF-24 using cassava starch (CS) as carbon source and the effects of different feeding methods were investigated in a fermenter. The initial shake flask experiments demonstrated the efficient production of ε-PL with CS, achieving the ε-PL production of 1.18 g/L. Subsequent investigations in the fermenter identified that the ideal pH was 3.8 during the ε-PL synthesis phase. Under this condition, the production of ε-PL reached 1.35 g/L. When the pH was maintained at 3.8, the investigation of improvement of feeding composition was carried out in a 5 L fermenter. The intermittent feeding containing CS, inorganic and organic nitrogen sources resulted in the maximum ε-PL production and dry cell weight (DCW) reaching 17.17 g/L and 42.73 g/L. Additionally, continuous feeding with the composition of CS, organic and inorganic nitrogen sources, and inorganic salts further increased ε-PL production and DCW to 27.56 g/L and 38.5 g/L. Summarily, the above results indicate that the fermentation using low-cost CS and continuous feeding strategy with whole medium composition can provide a beneficial reference for the efficient production of ε-PL.

9.
Sheng Wu Gong Cheng Xue Bao ; 40(8): 2513-2527, 2024 Aug 25.
Artículo en Chino | MEDLINE | ID: mdl-39174468

RESUMEN

L-lysine is an essential amino acid with broad applications in the animal feed, human food, and pharmaceutical industries. The fermentation production of L-lysine by Escherichia coli has limitations such as poor substrate utilization efficiency and low saccharide conversion rates. We deleted the global regulatory factor gene mlc and introduced heterologous genes, including the maltose phosphotransferase genes (malAP) from Bacillus subtilis, to enhance the use efficiency of disaccharides and trisaccharides. The engineered strain E. coli XC3 demonstrated improved L-lysine production, yield, and productivity, which reached 160.00 g/L, 63.78%, and 4.44 g/(L‧h), respectively. Furthermore, we overexpressed the glutamate dehydrogenase gene (gdhA) and assimilated nitrate reductase genes (BsnasBC) from B. subtilis, along with nitrite reductase genes (EcnirBD) from E. coli, in strain E. coli XC3. This allowed the construction of E. coli XC4 with a nitrate assimilation pathway. The L-lysine production, yield, and productivity of E. coli XC4 were elevated to 188.00 g/L, 69.44%, and 5.22 g/(L‧h), respectively. After optimization of the residual sugar concentration and carbon to nitrogen ratio, the L-lysine production, yield, and productivity were increased to 204.00 g/L, 72.32%, and 5.67 g/(L‧h), respectively, in a 5 L fermenter. These values represented the increases of 40.69%, 20.03%, and 40.69%, respectively, compared with those of the starting strain XC1. By engineering the substrate utilization pathway, we successfully constructed a high-yield L-lysine producing strain, laying a solid foundation for the industrial production of L-lysine.


Asunto(s)
Bacillus subtilis , Escherichia coli , Fermentación , Lisina , Ingeniería Metabólica , Escherichia coli/genética , Escherichia coli/metabolismo , Lisina/biosíntesis , Lisina/metabolismo , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Glutamato Deshidrogenasa/metabolismo , Glutamato Deshidrogenasa/genética , Nitrato-Reductasa/genética , Nitrato-Reductasa/metabolismo
10.
Int J Biol Macromol ; 277(Pt 2): 134208, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39069059

RESUMEN

Antibacterial hydrogels as burn wound dressings are capable of efficaciously defending against bacterial infection and accelerating burn wound healing. Thus far, a large plethora of antibacterial hydrogels have adopted numerous components and intricate preparation processes, yet restricting their practical industrialization applications. Simple and effective preparation methods of antibacterial hydrogels are hence urgently needed. Herein, an easy but efficacious strategy with the employment of two natural products pullulan and ε-poly-l-lysine (ε-PL) was designed to fabricate composite antibacterial hydrogels for burn wound healing for the first time. The hydrogel crosslinking networks were formed through amidation reactions between carboxylated pullulan derivative (CP) and ε-poly-l-lysine hydrochloride (ε-PL·HCl). The resulting hydrogels possessed high transparency, porous structures, tunable gelation time and gel content, relatively low swelling ratios, appropriate self-degradability, proper mechanical properties, strong in vitro bacteriostatic activities, non-cytotoxicity, capacities of facilitating cell migration and excellent hemocompatibility. In the infected burn model of mice, the hydrogels were observed to display prominent in vivo antibacterial activities and enable the acceleration of burn wound healing. We opine the simply and effectively prepared antibacterial hydrogels as promising dressings for burn wound recovery have broad industrialization prospects.


Asunto(s)
Antibacterianos , Quemaduras , Glucanos , Hidrogeles , Polilisina , Cicatrización de Heridas , Hidrogeles/química , Hidrogeles/farmacología , Glucanos/química , Glucanos/farmacología , Polilisina/química , Polilisina/farmacología , Cicatrización de Heridas/efectos de los fármacos , Antibacterianos/farmacología , Antibacterianos/química , Animales , Quemaduras/tratamiento farmacológico , Quemaduras/terapia , Ratones , Productos Biológicos/química , Productos Biológicos/farmacología , Staphylococcus aureus/efectos de los fármacos , Humanos , Pruebas de Sensibilidad Microbiana
11.
Biomolecules ; 14(7)2024 Jun 25.
Artículo en Inglés | MEDLINE | ID: mdl-39062465

RESUMEN

Safe and eco-friendly preservatives are crucial to preventing food spoilage and illnesses, as foodborne diseases caused by pathogens result in approximately 600 million cases of illness and 420,000 deaths annually. ε-Poly-L-lysine (ε-PL) is a novel food preservative widely used in many countries. However, its commercial application has been hindered by high costs and low production. In this study, ε-PL's biosynthetic capacity was enhanced in Streptomyces albulus WG608 through metabolic engineering guided by multi-omics techniques. Based on transcriptome and metabolome data, differentially expressed genes (fold change >2 or <0.5; p < 0.05) and differentially expressed metabolites (fold change >1.2 or <0.8) were separately subjected to gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. The integrative analysis of transcriptome, metabolome, and overexpression revealed the essential roles of isocitrate lyase, succinate dehydrogenase, flavoprotein subunit, diaminopimelate dehydrogenase, polyphosphate kinase, and polyP:AMP phosphotransferase in ε-PL biosynthesis. Subsequently, a strain with enhanced ATP supply, L-lysine supply, and ε-PL synthetase expression was constructed to improve its production. Finally, the resulting strain, S. albulus WME10, achieved an ε-PL production rate of 77.16 g/L in a 5 L bioreactor, which is the highest reported ε-PL production to date. These results suggest that the integrative analysis of the transcriptome and metabolome can facilitate the identification of key pathways and genetic elements affecting ε-PL synthesis, guiding further metabolic engineering and thus significantly enhancing ε-PL production. The method presented in this study could be applicable to other valuable natural antibacterial agents.


Asunto(s)
Ingeniería Metabólica , Polilisina , Streptomyces , Streptomyces/metabolismo , Streptomyces/genética , Ingeniería Metabólica/métodos , Polilisina/biosíntesis , Polilisina/metabolismo , Metaboloma , Transcriptoma , Metabolómica/métodos , Multiómica
12.
J Mech Behav Biomed Mater ; 157: 106646, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38981181

RESUMEN

Graphene oxide (GO) exhibits excellent mechanical strength and modulus. However, its effectiveness in mechanically reinforcing polymer materials is limited due to issues with interfacial bonding and dispersion arising from differences in the physicochemical properties between GO and polymers. Surface modification using coupling agents is an effective method to improve the bonding problem between polymer and GO, but there may be biocompatibility issues when used in the biomedical field. In this study, the biomolecule L-lysine, was applied to improve the interfacial bonding and dispersion of GO in polylactic acid (PLA) without compromising biocompatibility. The PLA/L-lysine-modified GO (PLA/L-GO) bone scaffold with triply periodic minimal surface (TPMS) structure was prepared using fused deposition modeling (FDM). The FTIR results revealed successful grafting of L-lysine onto GO through the reaction between their -COOH and -NH2 groups. The macroscopic and microscopic morphology characterization indicated that the PLA/L-GO scaffolds exhibited an characteristics of dynamic diameter changes, with good interlayer bonding. It was noteworthy that the L-lysine modification promoted the dispersion of GO and the interfacial bonding with the PLA matrix, as characterized by SEM. As a result, the PLA/0.1L-GO scaffold exhibited higher compressive strength (13.2 MPa) and elastic modulus (226.8 MPa) than PLA/0.1GO. Moreover, PLA/L-GO composite scaffold exhibited superior biomineralization capacity and cell response compared to PLA/GO. In summary, L-lysine not only improved the dispersion and interfacial bonding of GO with PLA, enhancing the mechanical properties, but also improved the biological properties. This study suggests that biomolecules like L-lysine may replace traditional modifiers as an innovative bio-modifier to improve the performance of polymer/inorganic composite biomaterials.


Asunto(s)
Grafito , Lisina , Ensayo de Materiales , Fenómenos Mecánicos , Poliésteres , Impresión Tridimensional , Andamios del Tejido , Poliésteres/química , Andamios del Tejido/química , Porosidad , Grafito/química , Lisina/química , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Animales
13.
Polymers (Basel) ; 16(14)2024 Jul 17.
Artículo en Inglés | MEDLINE | ID: mdl-39065363

RESUMEN

Polyamide-6 (PA) is a popular textile polymer having desirable mechanical and thermal properties, chemical stability, and biocompatibility. However, PA nanofibers are prone to bacterial growth and user discomfort. ε-Poly-L-lysine (PL) is non-toxic, antimicrobial, and hydrophilic but lacks spinnability due to its low molecular weight. Given its similar backbone structure to PA, with an additional amino side chain, PL was integrated with PA to develop multifunctional nanofibers. This study explores a simple, scalable method by which to obtain PL nanofibers by utilizing the structurally similar PA as the base. The goal was to enhance the functionality of PA by addressing its drawbacks. The study demonstrates spinnability of varying concentrations of PL with base PA while exploring compositions with higher PL concentrations than previously reported. Electrospinning parameters were studied to optimize the nanofiber properties. The effects of PL addition on morphology, hydrophilicity, thermal stability, mechanical performance, and long-term antimicrobial activity of nanofibers were evaluated. The maximum spinnable concentration of PL in PA-based nanofibers resulted in super hydrophilicity (0° static water contact angle within 10 s), increased tensile strength (1.02 MPa from 0.36 MPa of control), and efficient antimicrobial properties with long-term stability. These enhanced characteristics hold promise for the composite nanofiber's application in medical and protective textiles.

14.
J Food Sci ; 89(7): 4298-4311, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38957101

RESUMEN

This study explored the mechanism of l-lysine intervention in wheat gluten protein (WG) gel formation under a microwave (MW) field. The results showed that the MW treatment had higher ζ-potential values at the same heating rate. After adding l-lysine, the solution conductivity and dielectric loss were significantly increased. Moreover, the WG gel strength enhanced 4.40% under the MW treatment. The Fourier spectra showed that the α-helix content was decreased 13.78% with the addition of lysine. The ultraviolet absorption spectra and fluorescence spectra indicated that MW irradiation impacted the interactions between WG molecules more effectively than the water bath heating, promoting the denaturation and unfolding of the protein structure. In addition, scanning electron microscopy analysis showed that the incorporation of lysine promoted an ordered network structure formation of the protein, which enhanced the gel properties. This indicated that the zwitterion of l-lysine played a regulatory role in the aggregation of proteins in the MW field.


Asunto(s)
Glútenes , Lisina , Microondas , Triticum , Lisina/química , Triticum/química , Glútenes/química , Agregado de Proteínas , Proteínas de Plantas/química , Calor , Geles/química
15.
Sci Rep ; 14(1): 15181, 2024 07 02.
Artículo en Inglés | MEDLINE | ID: mdl-38956295

RESUMEN

Human norovirus (HuNoV) is an enteric infectious pathogen belonging to the Caliciviridae family that causes occasional epidemics. Circulating alcohol-tolerant viral particles that are readily transmitted via food-borne routes significantly contribute to the global burden of HuNoV-induced gastroenteritis. Moreover, contact with enzymes secreted by other microorganisms in the environment can impact the infectivity of viruses. Hence, understanding the circulation dynamics of Caliciviridae is critical to mitigating epidemics. Accordingly, in this study, we screened whether environmentally abundant secretase components, particularly proteases, affect Caliciviridae infectivity. Results showed that combining Bacillaceae serine proteases with epsilon-poly-L-lysine (EPL) produced by Streptomyces-a natural antimicrobial-elicited anti-Caliciviridae properties, including against the epidemic HuNoV GII.4_Sydney_2012 strain. In vitro and in vivo biochemical and virological analyses revealed that EPL has two unique synergistic viral inactivation functions. First, it maintains an optimal pH to promote viral surface conformational changes to the protease-sensitive structure. Subsequently, it inhibits viral RNA genome release via partial protease digestion at the P2 and S domains in the VP1 capsid. This study provides new insights regarding the high-dimensional environmental interactions between bacteria and Caliciviridae, while promoting the development of protease-based anti-viral disinfectants.


Asunto(s)
Bacillaceae , Polilisina , Serina Proteasas , Streptomyces , Streptomyces/enzimología , Polilisina/farmacología , Polilisina/química , Polilisina/metabolismo , Serina Proteasas/metabolismo , Bacillaceae/enzimología , ARN Viral/genética , ARN Viral/metabolismo , Humanos , Genoma Viral , Animales , Norovirus/efectos de los fármacos , Norovirus/genética , Inactivación de Virus/efectos de los fármacos , Caliciviridae/genética , Antivirales/farmacología
16.
J Dent Sci ; 19(3): 1748-1757, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-39035281

RESUMEN

Background/purpose: Helicobacter pylori has been found to be related to periodontitis, and the oral cavity has been considered a reservoir for H. pylori gastritis infection. Thus, this study evaluated the effect of mouthwash containing poly l-Lysine and glycerol monolaurate on inhibiting H. pylori growth, biofilm formation, cell cytotoxicity, adhesion ability, cagA mRNA expression, and pro-inflammatory cytokines stimulated by H. pylori. Materials and methods: Nineteen H. pylori strains were isolated from the oral cavity. The effectiveness of mouthwash containing poly l-Lysine and glycerol monolaurate was examined for its ability to inhibit H. pylori growth and biofilm formation and was tested for cell viability in oral epithelial cells (H357), gastric adenocarcinoma cells (AGS), and periodontal ligament cells (PDL). Additionally, the mouthwash was tested for reducing cagA mRNA expression, adhesion ability to H357 and AGS cells, and pro-inflammatory cytokines stimulated with H. pylori in AGS and PDL cells. Results: The mouthwash containing poly l-Lysine and glycerol monolaurate could eradicate the biofilm by 14.9-19.9% after incubation at 5 min, and cell viability revealed 77.2, 79.8, and 100.0% for AGS, H357, and PDL cells, respectively. Moreover, the mouthwash containing poly l-Lysine and glycerol monolaurate could down-regulate cagA mRNA expression, reduce adhesion of H. pylori by approximately 9.5-47.8% for H357 cells and 24.5-62.9% for AGS cells, and decrease pro-inflammatory cytokines, especially interleukin-8, stimulated with H. pylori. Conclusion: Mouthwash containing poly l-Lysine and glycerol monolaurate could inhibit H. pylori growth and reduce their virulence expression. The mouthwash also revealed low cytotoxicity to oral and gastric cells.

17.
Enzyme Microb Technol ; 179: 110467, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38852284

RESUMEN

ε-Poly-l-lysine (ε-PL), a natural food preservative with various advantages, is primarily produced by Streptomyces. It has attracted considerable attentions for the outstanding antibacterial activity, safety, heat stability, water solubility and other remarkable properties. In this study, a food-grade recombinant Bacillus subtilis was constructed for the biocatalysis of ε-PL. Firstly, the d-alanine racemase gene (alrA) was deleted from the genome of Bacillus subtilis 168 to construct an auxotrophic B. subtilis 168 (alrA-). Based on the shuttle plasmid pMA5, a food-grade plasmid pMA5a was constructed by replacing the genes of kanamycin resistance (Kanr) and ampicillin resistance (Ampr) with alrA and the gene encoding α-peptide of ß-galactosidase (lacZα), respectively. Subsequently, codon-optimized ε-PL synthase gene (pls) and P-pls were ligated into pMA5a and transformed in E. coli DH5α and expressed in B. subtilis 168 (alrA-). Finally, the whole-cell biocatalysis conditions for ε-PL production by B. subtilis 168 (alrA-)/pMA5a-pls were optimized, and the optimal conditions were 30°C, pH 4, l-lysine concentration of 0.6 g/L, bacterial concentration of 15 % (w/v) and a catalytic time of 7 h. The ε-PL production reached a maximum of 0.33 ± 0.03 g/L. The product was verified to be ε-PL by HPLC and tricine-SDS-PAGE. The information obtained in this study shows critical reference for the food-grade heterologous expression of ε-PL.


Asunto(s)
Bacillus subtilis , Biocatálisis , Polilisina , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Bacillus subtilis/enzimología , Polilisina/metabolismo , Polilisina/biosíntesis , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Plásmidos/genética , Escherichia coli/genética , Escherichia coli/metabolismo
18.
ACS Appl Mater Interfaces ; 16(25): 31922-31935, 2024 Jun 26.
Artículo en Inglés | MEDLINE | ID: mdl-38874539

RESUMEN

Poly-l-lysine (PLL) and Matrigel, both classical coating materials for culture substrates in neural stem cell (NSC) research, present distinct interfaces whose effect on NSC behavior at cellular and molecular levels remains ambiguous. Our investigation reveals intriguing disparities: although both PLL and Matrigel interfaces are hydrophilic and feature amine functional groups, Matrigel stands out with lower stiffness and higher roughness. Based on this diversity, Matrigel surpasses PLL, driving NSC adhesion, migration, and proliferation. Intriguingly, PLL promotes NSC differentiation into astrocytes, whereas Matrigel favors neural differentiation and the physiological maturation of neurons. At the molecular level, Matrigel showcases a wider upregulation of genes linked to NSC behavior. Specifically, it enhances ECM-receptor interaction, activates the YAP transcription factor, and heightens glycerophospholipid metabolism, steering NSC proliferation and neural differentiation. Conversely, PLL upregulates genes associated with glial cell differentiation and amino acid metabolism and elevates various amino acid levels, potentially linked to its support for astrocyte differentiation. These distinct transcriptional and metabolic activities jointly shape the divergent NSC behavior on these substrates. This study significantly advances our understanding of substrate regulation on NSC behavior, offering novel insights into optimizing and targeting the application of these surface coating materials in NSC research.


Asunto(s)
Diferenciación Celular , Proliferación Celular , Colágeno , Combinación de Medicamentos , Laminina , Células-Madre Neurales , Polilisina , Proteoglicanos , Polilisina/química , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Células-Madre Neurales/efectos de los fármacos , Laminina/química , Laminina/farmacología , Colágeno/química , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Proteoglicanos/química , Proteoglicanos/farmacología , Animales , Adhesión Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Ratones
19.
ACS Appl Bio Mater ; 7(7): 4654-4663, 2024 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-38867502

RESUMEN

The 5-nitroimidazole (5-NI) class of antibiotics, such as metronidazole, ornidazole, secnidazole, and tinidazole, are widely used to prevent bacterial infection in humans and livestock industries. However, their overuse contaminates the farmed animal products and water bodies. Hence, a selective, sensitive, and cost-effective method to detect 5-NI antibiotics is the need of the hour. Herein, we report a rapid, inexpensive, and efficient sensing system to detect 5-NI drugs using an as-prepared solution of ε-poly-l-lysine (ε-PL), a naturally occurring and biodegradable homopolypeptide that has an intrinsic fluorescence via clustering-triggered emission. The low nanomolar detection limit (3.25-3.97 nM) for the aforementioned representative 5-NI drugs highlights the sensitivity of the system, outperforming most of the reported sensors alike. The resulting fluorescence quenching was found to be static in nature. Importantly, excellent recovery (100.26-104.41%) was obtained for all real samples and animal products tested. Visual detection was demonstrated by using paper strips and silica gel for practical applications. Furthermore, ε-PL could detect 5-NI antibiotics in living 3T3-L1 mouse fibroblast cells via cellular imaging. Taken together, the present work demonstrates the detection of 5-NI antibiotics using a biocompatible natural polypeptide, ε-PL, and represents a simple and inexpensive analytical tool for practical application.


Asunto(s)
Antibacterianos , Nitroimidazoles , Polilisina , Animales , Polilisina/química , Antibacterianos/química , Antibacterianos/farmacología , Antibacterianos/análisis , Ratones , Nitroimidazoles/química , Nitroimidazoles/análisis , Materiales Biocompatibles/química , Ensayo de Materiales , Tamaño de la Partícula , Fluorescencia , Estructura Molecular , Péptidos/química , Colorantes Fluorescentes/química , Imagen Óptica , Supervivencia Celular/efectos de los fármacos
20.
Pestic Biochem Physiol ; 202: 105959, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38879341

RESUMEN

ε-Poly-l-lysine (ε-PL) is an effective antimicrobial peptide for controlling fungal plant diseases, exhibiting significant antifungal activity and safety. Despite its known efficacy, the potential of ε-PL in combating plant bacterial diseases remains underexplored. This study evaluated the effectiveness of ε-PL and its nanomaterial derivative in managing tomato bacterial spot disease caused by Pseudomonas syringae pv. tomato. Results indicated that ε-PL substantially inhibited the growth of Pseudomonas syringae pv. tomato. Additionally, when ε-PL was loaded onto attapulgite (encoded as ATT@PL), its antibacterial effect was significantly enhanced. Notably, the antibacterial efficiency of ATT@PL containing 18.80 µg/mL ε-PL was even close to that of 100 µg/mL pure ε-PL. Further molecular study results showed that, ATT@PL stimulated the antioxidant system and the salicylic acid signaling pathway in tomatoes, bolstering the plants disease resistance. Importantly, the nanocomposite demonstrated no negative effects on both seed germination and plant growth, indicating its safety and aligning with sustainable agricultural practices. This study not only confirmed the effectiveness of ε-PL in controlling tomato bacterial spot disease, but also introduced an innovative high antibacterial efficiency ε-PL composite with good bio-safety. This strategy we believe can also be used in improving other bio-pesticides, and has high applicability in agriculture practice.


Asunto(s)
Antibacterianos , Enfermedades de las Plantas , Polilisina , Pseudomonas syringae , Compuestos de Silicona , Solanum lycopersicum , Pseudomonas syringae/efectos de los fármacos , Solanum lycopersicum/microbiología , Polilisina/farmacología , Polilisina/química , Antibacterianos/farmacología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Compuestos de Silicona/farmacología , Compuestos de Silicona/química , Compuestos de Magnesio
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