Search Results (64)

This study aimed to investigate the efficacy of a composite coating composed of 150 mg/L ε-Poly-L-lysine (ε-PL) and 5 g/L chitosan (CTS) in extending the shelf life and maintaining the postharvest quality of fresh Tremella fuciformis. Freshly harvested T. fuciformis were treated by surface spraying, with distilled water serving as the control. The effects of the coating on storage quality, physicochemical properties, reactive oxygen species (ROS) metabolism, and membrane lipid metabolism were evaluated during storage at (25 ± 1) °C. The results showed that the ε-PL/CTS composite coating significantly retarded quality deterioration, as evidenced by reduced weight loss, maintained whiteness and color, and higher retention of soluble sugars, soluble solids, and soluble proteins. The coating also effectively limited water migration and loss. Mechanistically, the coated T. fuciformis exhibited enhanced antioxidant capacity, characterized by increased superoxide anion (O₂⁻) resistance capacity, higher activities of antioxidant enzymes (SOD, CAT, APX), and elevated levels of non-enzymatic antioxidants (AsA, GSH). This led to a significant reduction in malondialdehyde (MDA) accumulation, alongside improved DPPH radical scavenging activity and reducing power. Furthermore, the ε-PL/CTS coating preserved cell membrane integrity by inhibiting the activities of lipid-degrading enzymes (lipase, LOX, PLD), maintaining higher levels of key phospholipids (phosphatidylinositol and phosphatidylcholine), delaying phosphatidic acid accumulation, and consequently reducing cell membrane permeability. In conclusion, the ε-PL/CTS composite coating effectively extends the shelf life and maintains the quality of postharvest T. fuciformis by modulating ROS metabolism and preserving membrane lipid homeostasis. This study provides a theoretical basis and a practical approach for the quality control of fresh T. fuciformis. Full article

This study aimed to investigate the mechanism underlying the synergistic antimicrobial effect of ferulic acid (FA) and ε-polylysine hydrochloride (PL) on Shewanella putrefaciens (S. putrefaciens) and their application on crayfish (Procambarus clarkii). The treatment with FA and PL exhibited a strong synergistic inhibitory effect against S. putrefaciens. The combination of 1/4 Minimum Inhibitory Concentration (MIC) FA and 1/4 MIC PL was the most effective, damaging the cell structure and inhibiting the growth of S. putrefaciens. Plasma-activated water (PAW) can induce microbial inactivation through physical action. In addition, treatments with FA, PL, PAW, PAW-FA, PAW-PL, and PAW + PL-FA substantially decreased total viable counts (TVCs), total volatile base nitrogen (TVB-N), the thiobarbituric acid value (TBA), and the juice loss rate of crayfish, with FA-PL showing the best effect. This study confirmed the antimicrobial efficacy of PL, FA, and PAW, indicating their potential as effective preservatives for controlling spoilage in freshwater crustaceans. Full article

Wound healing is a complex biological process involving inflammation, proliferation, and remodeling phases. Effective healing is essential for maintaining skin integrity, driving the need for advanced materials like hydrogels, known for their high water retention and tunable mechanical properties. In this study, we synthesized a biocompatible composite hydrogel composed of γ-polyglutamic acid (γ-PGA) and ε-polylysine (ε-PL) through a Schiff base reaction, forming a stable crosslinked network. Its physicochemical properties, including rheological behavior and swelling capacity, were systematically evaluated. Biocompatibility was assessed via in vitro hemolysis and cytotoxicity assays, and in vivo testing was performed using a full-thickness skin defect model in Sprague Dawley (SD) rats to evaluate wound-healing efficacy. The PGA-PL hydrogel demonstrated excellent physicochemical properties, with a maximum swelling ratio of 65.6%, and biocompatibility as evidenced by low hemolysis rates (<5%) and high cell viability (>80%). It promoted wound healing by inhibiting the inflammatory response, reducing levels of the inflammatory cytokine IL-6, enhancing angiogenesis, and accelerating collagen deposition. The hydrogel showed complete biodegradation within 21 days in vivo without inducing a significant inflammatory response and significantly accelerated wound healing, achieving an 86% healing rate within 7 days compared to 67% in the control group. The PGA-PL composite hydrogel exhibits excellent mechanical strength and biocompatibility, and its effective wound-healing capabilities lay the groundwork for future development and optimization in various tissue engineering applications. Full article

Guava fruit is susceptible to decay, leading to losses in storability and quality. ε-Poly-l-lysine (ε-PL) is a safe antimicrobial polypeptide that has proven to be effective in preserving produce’s quality. In the present research, ε-PL, at multiple concentrations (1, 2 and 4 g/L), was adopted to treat guavas, and the fruit were stored at 25 °C for 15 d. The results indicated that ε-PL retarded the guava storability decline and enhanced its quality. Treated guavas had a better appearance, as well as the lower disease index, relative electrolytic leakage, weight loss, respiration intensity, a* and b* values and reducing sugar content. They also showed higher firmness, commercially acceptable fruit rate, titratable acidity, L* value, total soluble sugar, vitamin C and sucrose levels. The optimal concentration of ε-PL was determined to be 2 g/L. Furthermore, compared to control guavas, fruit treated with 2 g/L ε-PL exhibited lower levels of superoxide anion, hydrogen peroxide and malondialdehyde but higher antioxidant enzyme activities in terms of ascorbate peroxidase, peroxidase, catalase and superoxide dismutase. These findings suggested that ε-PL raised the antioxidant enzyme activities to enhance the fruit’s antioxidant ability. This, in turn, reduced the reactive oxygen species levels and lipid peroxidation, ultimately improving the guava’s quality. Consequently, ε-PL is of practical significance for commercial application as it suppresses decay and stabilizes the quality of guavas, enhancing their postharvest marketability. Full article

Crab roe sauce (CRS) is prone to spoilage due to microbial contamination. Therefore, this study aimed to investigate the inhibitory effects and mechanisms of a mixed bio-preservative (0.025% ε-polylysine hydrochloride (ε-PL) + 0.01% nisin (NS) + 0.01% tea polyphenols (TPs)) on the specific spoilage bacteria (SSB) in CRS. First, the SSB in CRS were isolated and identified by 16S rRNA sequencing. Two isolates were selected as representative strains based on their enzymatic spoilage potential and spoilage capability in CRS. By comparing the inhibition zones, ε-PL, NS, and TPs were selected from five conventional bio-preservatives (ε-PL, NS, TPs, grape seed extract (GSE), and rosemary extract (RE)) to prepare the mixed bio-preservative. The results showed that the minimum inhibitory concentration (MIC) of the mixed bio-preservative against Bacillus pumilus and Bacillus subtilis was 56.3 µg/mL. The growth curves and cell viability tests revealed that the mixed bio-preservative reduced the viability of both strains. The conductivity, alkaline phosphatase activity, and nucleic acid and soluble protein leakage indicated that the mixed bio-preservative disrupted the integrity of the cell walls and membranes of the two isolates in a concentration-dependent manner. Scanning electron microscopy further confirmed the damage to the cell membranes of the two isolates by the mixed bio-preservative. Overall, the mixed bio-preservative exhibited excellently inhibitory effects on the SSB and could be a promising method for the preservation of CRS. Full article

ε-poly-l-lysine (ε-PL), a natural food preservative, has garnered widespread attention. It is mainly produced by Streptomyces albulus, but the production by wild-type strains fails to meet the demands of industrialization. To address this issue, adaptive laboratory evolution (ALE) was successfully employed in this study, subjecting S. albulus CICC 11022 to environmental stresses such as acidic pH and antibiotics (rifampicin, gentamicin, and streptomycin). As a result of ALE, an evolutionary strain S. albulus C214 was obtained, exhibiting an increase in ε-PL production and cell growth by 153.23% and 234.51%, respectively, as compared with the original strain. Genomic and metabolic analyses revealed that mutations occurred in genes responsible for transcriptional regulation, transporter, cell envelope, energy metabolism, and secondary metabolite synthesis, as well as the enrichment of metabolites involved in the biosynthesis of ε-PL. These findings hold great significance for elucidating the mechanism underlying ε-PL synthesis. Full article

ε-Poly-L-lysine (ε-PL) is a natural preservative that has excellent properties such as high safety, good antibacterial effect, wide antibacterial spectrum, and high temperature resistance compared to other food preservatives. At present, the main production strain of ε-PL is Streptomyces albulus (S. albulus). Due to the large amount of mycelium and by-products during fermentation, its production cost is much higher than other food preservatives, which seriously hinders the application of ε-PL in the food industry. Corynebacterium glutamicum (C. glutamicum) is a food safety strain that is widely used in the fermentation industry to produce various amino acids. Its ability to produce high amounts of L-lysine can provide sufficient precursor substances for the synthesis of ε-PL, making it an ideal strain for the heterologous expression of ε-PL synthase genes (pls). In this experiment, a recombinant C. glutamicum capable of synthesizing ε-PL and exhibiting certain physiological resistance to ε-PL was obtained by amplifying pls and heterologous expression in C. glutamicum for the first time. Further optimization of the fermentation temperature, initial pH, and inoculation amount of the recombinant strain resulted in an increase in the ε-PL fermentation yield from 0.12 g/L to 0.22 g/L. Finally, through adaptive evolution of the recombinant strain, the ε-PL tolerance of the recombinant strain was increased to 1.3 g/L, and the yield of ε-PL ultimately reached 0.34 g/L, which increased by 54.55% compared to the initial strain. The recombinant C. glutamicum constructed in this study can significantly shorten the fermentation cycle, reduce bacterial volume and the synthesis of secondary metabolites, which is beneficial for the separation and purification of products, thereby further reducing the production cost of ε-PL and accelerating the process of replacing chemical food preservatives with natural food preservatives. Full article

Hydrogels have emerged as promising biomaterials due to their excellent performance; however, their biocompatibility, biodegradability, and absorbability still require improvement to support a broader range of medical applications. This paper presents a new biofunctionalized hydrogel based on in situ crosslinking between maleimide-terminated four-arm-poly(ethylene glycol) (4–arm–PEG–Mal) and poly(ε-lysine) (ε–PL). The PEG/ε–PL hydrogels, named LG–n, were rapidly formed via amine/maleimide reaction by mixing 4–arm–PEG–Mal and ε–PL under physiological conditions. The corresponding dry gels (DLG–n) were obtained through a freeze-drying technique. ¹H NMR, FT–IR, and SEM were utilized to confirm the structures of 4–arm–PEG–Mal and LG–n (or DLG–n), and the effects of solid content on the physicochemical properties of the hydrogels were investigated. Although high solid content could increase the swelling ratio, all LG–n samples exhibited a low equilibrium swelling ratio of less than 30%. LG–7, which contained moderate solid content, exhibited optimal compression properties characterized by a compressive fracture strength of 45.2 kPa and a deformation of 69.5%. Compression cycle tests revealed that LG–n demonstrated good anti-fatigue performance. In vitro degradation studies confirmed the biodegradability of LG–n, with the degradation rate primarily governing the drug (ceftibuten) release efficiency, leading to a sustained release duration of four weeks. Cytotoxicity tests, cell survival morphology observation, live/dead assays, and hemolysis tests indicated that LG–n exhibited excellent cytocompatibility and low hemolysis rates (<5%). Furthermore, the broad-spectrum antibacterial activity of LG–n was verified by an inhibition zone method. In conclusion, the developed LG–n hydrogels hold promising applications in the medical field, particularly as drug sustained-release carriers and wound dressings. Full article

The post-harvest process is important to increasing the market value of limes and requires focus. During this process, limes are graded and categorized based on size, weight, and volume. Therefore, identifying efficient means of estimating these properties is very important and remains an open research area. This study applies the concept of computer vision based on traditional machine learning algorithms (partial least square regression (PLS), epsilon-support vector regression (ε-SVR), decision tree (DT), random forest (RF), adaptive boosting (AB), gradient boosting (GB), Bagging meta-estimator (BME), and extremely randomized trees (ERTs)) and pre-trained deep learning (InceptionV3, MoblieNetV2, ResNet50, and VGG-16) for estimating the weight and volume of limes. Our findings showed that the BME and ResNet50 could yield the highest performance for estimating the weight and volume of limes. The BME produced ${\mathrm{R}}_{\mathrm{test}}^2$ values of 0.954 and 0.882 for weight and volume, respectively, while the ${\mathrm{R}}_{\mathrm{test}}^2$ values of ResNet50 models were between 0.951 and 0.957 for weight and volume, respectively. This study concluded that computer vision based on both traditional machine learning and deep learning could be used to estimate the weight and volume of limes. The approach proposed in this study can be adopted for applications related to computer vision in the post-harvest process. Full article

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. Full article

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. Full article

Gray mold, caused by Botrytis cinerea, poses significant threats to various crops, while it can be remarkably inhibited by ε-poly-L-lysine (ε-PL). A previous study found that B. cinerea extracts could stimulate the ε-PL biosynthesis of Streptomyces albulus, while it is unclear whether the impact of the B. cinerea signal on ε-PL biosynthesis is direct or indirect. This study evaluated the role of elevated reactive oxygen species (ROS) in efficient ε-PL biosynthesis after B. cinerea induction, and its underlying mechanism was disclosed with a transcriptome analysis. The microbial call from B. cinerea could arouse ROS elevation in cells, which fall in a proper level that positively influenced the ε-PL biosynthesis. A systematic transcriptional analysis revealed that this proper dose of intracellular ROS could induce a global transcriptional promotion on key pathways in ε-PL biosynthesis, including the embden-meyerhof-parnas pathway, the pentose phosphate pathway, the tricarboxylic acid cycle, the diaminopimelic acid pathway, ε-PL accumulation, cell respiration, and energy synthesis, in which sigma factor HrdD and the transcriptional regulators of TcrA, TetR, FurA, and MerR might be involved. In addition, the intracellular ROS elevation also resulted in a global modification of secondary metabolite biosynthesis, highlighting the secondary signaling role of intracellular ROS in ε-PL production. This work disclosed the transcriptional mechanism of efficient ε-PL production that resulted from an intracellular ROS elevation after B. cinerea elicitors’ induction, which was of great significance in industrial ε-PL production as well as the biocontrol of gray mold disease. Full article

Several Pleurocordyceps species have been reported as hyperparasitic fungi. A new species, Pleurocordyceps fusiformispora, and a known species, Perennicordyceps elaphomyceticola, are described here based on morphology and phylogenetic evidence from six genes (ITS, SSU, LSU, TET1-α, RPB1, and RPB2). Pl. fusiformispora differed from the other Pleurocordyceps species by producing flaky colonies, ovoid or elliptic α-conidia, and fusiform or long fusiform β-conidia. Both full genomes of Pe. elaphomyceticola and Pl. fusiformispora were sequenced, annotated, and compared. The antiSMASH and local BLAST analyses revealed significant differences in the number and types of putative secondary metabolite biosynthetic gene clusters, i.e., NPPS, PKS, and hybrid PKS–NRPS domains, between the two species. In addition, the putative BGCs of six compounds, namely ε-poly lysine, 4-epi-15-epi-brefeldin A, Monorden D/monocillin IV/monocillin VII/pochonin M/monocillin V/monocillin II, Tolypyridone, Piperazine, and Triticone DABFC, were excavated in the present study. This study motivates the use of heterologous expression and gene knockout methods to discover novel biologically active SMs from Polycephalomycetaceae. Full article

Pseudomonas aeruginosa and Klebsiella pneumoniae are notorious for their resistance to antibiotics and propensity for biofilm formation, posing significant threats to human health. Epsilon-poly-L-lysine (ε-PL) emerges as a naturally occurring antimicrobial poly(amino acid), which positions it as a prospective agent for addressing challenges linked to multidrug resistance. ε-PL symbolizes a promising avenue in the pursuit of efficacious therapeutic strategies and warrants earnest consideration within the realm of clinical treatment. Thus, our objective was to determine the antibiotic susceptibility profiles of 38 selected P. aeruginosa and ESBL-producing K. pneumoniae clinical isolates and determine the ability of ε-PL to inhibit biofilm formation. After PCR analysis, detection of genes related to β-lactamases was observed among the selected isolates of P. aeruginosa [bla_SPM (35.7%), bla_KPC (35.7%), bla_SHV (14.3%), bla_CTX-M (14.3%), bla_OXA (14.3%), bla_TEM (7.1%), bla_PER (7.1%), bla_VIM (7.1%), and bla_VIM-2 (7.1%)] and K. pneumoniae [bla_CTX-M (91.7%), bla_TEM (83.3%), bla_KPC (16.7%), bla_NDM (12.5%), and bla_OXA (4.2%)]. The results of testing the activity of ε-PL against the clinical isolates showed relatively high minimum inhibitory concentrations (MICs) for the P. aeruginosa (range: 8–64 µg/mL) and K. pneumoniae isolates (range: 16–32 µg/mL). These results suggest the need for prior optimization of ε-PL concerning its viability as an alternative to antibiotics for treating infections caused by P. aeruginosa and K. pneumoniae of clinical origin. It is noteworthy that, in the context of a low antibiotic discovery rate, ε-PL could play a significant role in this quest, considering its low toxicity and the unlikely development of resistance. Upon exposure to ε-PL, P. aeruginosa and K. pneumoniae isolates exhibited a reduction in biofilm production, with ε-PL concentration showing an inverse relationship, particularly in isolates initially characterized as strong or moderate producers, indicating its potential as a natural antimicrobial agent with further research needed to elucidate optimal concentrations and application methods across different bacterial species. Further research is needed to optimize its use and explore its potential in various applications. Full article

Freshly harvested Tremella fuciformis contains high water content with an unprotected outer surface and exhibits high respiration rates, which renders it prone to moisture and nutrient loss, leading to decay during storage. Our research utilized ε-poly-L-lysine (ε-PL) and chitosan as a composite coating preservative on fresh T. fuciformis. The findings revealed that the ε-PL + chitosan composite coating preservative effectively delayed the development of diseases and reduced weight loss during storage compared to the control group. Furthermore, this treatment significantly decreased the respiration rate of T. fuciformis and the activity of respiratory metabolism-related enzymes, such as alternative oxidase (AOX), cytochrome c oxidase (CCO), succinic dehydrogenase (SDH), 6-phosphogluconate dehydrogenase, and glucose-6-phosphate dehydrogenase (6-PGDH and G-6-PDH). Additionally, the composite coating preservative also delayed the depletion of ATP and ADP and maintained higher levels of the energy charge while preserving low levels of AMP. It also sustained heightened activities of Mg²⁺-ATPase, Ca²⁺-ATPase, and H⁺-ATPase enzymes. These results demonstrate that utilizing the ε-PL + chitosan composite coating preservative can serve as a sufficiently safe and efficient method for prolonging the shelf life of post-harvest fresh T. fuciformis. Full article