Search Results (8)

The fibrin matrix of thrombi is intertwined with neutrophil extracellular traps (NETs) containing histones that render resistance to fibrinolysis. During NET formation, histones are citrullinated. Our study addresses the question of whether citrullination modifies the fibrin-stabilizing effects of histones. We studied the structure and viscoelastic properties of fibrin formed in the presence of native or citrullinated H1 and core histones by scanning electron microscopy, clot permeation, and oscillation rheometry. The kinetics of fibrin formation and its dissolution were followed by turbidimetry and thromboelastometry. Co-polymerizing H1 with fibrin enhanced the mechanical strength of the clots, thickened the fibrin fibers, and enlarged the gel pores. In contrast, the addition of core histones resulted in a reduction in the fiber diameter, and the pores were only slightly larger, whereas the mechanical stability was not modified. Plasmin-mediated fibrinogen degradation was delayed by native and citrullinated core histones, but not by H1, and the action of des-kringle1-4-plasmin was not affected. Plasmin-mediated fibrinolysis was inhibited by native and citrullinated core histones, and this effect was moderated when the kringle domains of plasmin were blocked or deleted. These findings suggest that in NET-containing thrombi that are rich in core histones, alternative fibrinolytic enzymes lacking kringle domains are more efficient lytic agents than the classic plasmin-dependent fibrinolysis. Full article

Maize (Zea mays L.) silage in the irrigated and flat areas of Italy represents the most important large ruminant feed crop due to the high dry matter yield and nutritive value per hectare. The aim of the investigation was to evaluate the chemical composition and the in vitro fermentation patterns of five maize varieties (Tiesto, R700 1, MAS 78.T, DKC 7074 and KWS Kantico) freshly chopped and preserved via ensiling. The results indicated that the chemical composition was not significantly different among varieties. The substrates were incubated for 72 h with buffered rumen fluid collected from cow. The ensiling process slightly reduced gas production and fermentation kinetics, likely due to the consumption of soluble sugars during fermentation. Organic matter loss (OM loss) differed significantly (p < 0.01) among varieties in ensiled maize, correlating with their neutral detergent fiber (NDF) content. While total volatile fatty acid (VFA) production showed no significant differences between varieties, the buffer capacity ratio (BCR), an indicator of protein degradation, varied significantly. Ammonia production (NH₃) was significantly higher in ensiled samples, supporting previous findings that ensiling increases non-protein nitrogen (NPN) due to microbial proteolysis and plant enzyme activity. The gas production profiles and fermentation rates over time showed minor differences between fresh and ensiled samples, with fresh material exhibiting faster fermentation kinetics due to the presence of soluble sugars. These findings highlight the importance of evaluating maize silage quality to optimize ruminant nutrition and feed efficiency. Full article

The increasing generation of agro-industrial waste and its improper disposal have raised significant environmental concerns, highlighting the urgent need for sustainable alternatives which would repurpose these materials. In this context, enzymes such as endoglucanase play a critical role in degrading lignin–cellulose biomass by catalyzing the breakdown of β-1,4-glycosidic bonds in cellulose, thereby converting it into fermentable sugars with diverse industrial applications. This study aimed to investigate the production, purification, and characterization of an endoglucanase produced by the fungus Pleurotus djamor PLO13, using coconut fiber, sugarcane bagasse, wheat bran, and pineapple crown as substrates. Endoglucanase activity was measured by the Miller method (1959), using 2% (w/v) carboxymethyl cellulose (CMC) as substrate. Solid-state fermentation (SSF) was found to be highly efficient for enzyme synthesis, with wheat bran emerging as the most effective substrate, yielding an enzyme production of 7.19 U after 120 h of cultivation. The endoglucanase was purified through ethanol precipitation and ion-exchange chromatography using DEAE-Sepharose, achieving a recovery rate of 110%, possibly due to removal of inhibitors present in the crude extract. The purified enzyme exhibited stability across a broad pH range and thermostability, with optimal activity at pH 5.0 and 50 °C. Furthermore, the enzyme was activated by EDTA, Mn²⁺, and Ca²⁺, while being inhibited by Mg²⁺. Notably, the enzyme demonstrated halotolerance, with activity increasing by 60% upon the addition of 3 M NaCl. Kinetic analysis revealed that the purified enzyme showed affinity to the CMC substrate at the analyzed parameters (pH 5.0 and 50 °C), with Km and Vmax values of 0.0997 mg/mL and 112.2 µg/min/mL, respectively. These findings suggest that the endoglucanase from P. djamor PLO13 has promising potential for biotechnological applications, underscoring the feasibility of the use of lignocellulosic waste as sustainable substrates in industrial processes. Full article

The escalating demand for sustainable materials has been fueling the rapid proliferation of the biopolymer market. Biodegradable polymers within natural habitats predominantly undergo degradation mediated by microorganisms. These microorganisms secrete enzymes that cleave long-chain polymers into smaller fragments for metabolic assimilation. This review is centered around dissecting the degradation mechanisms of specific biodegradable polymers, namely PLA, starch-based polymers, and plant fiber-based polymers. Recent investigations have unveiled that PLA exhibits augmented biocompatibility when combined with HA, and its degradation is subject to the influence of enzymatic and abiotic determinants. In the case of starch-based polymers, chemical or physical modifications can modulate their degradation kinetics, as evidenced by Wang et al.’s superhydrophobic starch-based nanocomposite cryogel. For plant fiber-based polymers, the effects of temperature, humidity, and cellulose degradation on their properties, along with the implications of various treatments and additives, are probed, as exemplified by Liu et al.’s study on jute/SiO₂/PP composites. Specifically, with respect to PLA, the polymerization process and the role of catalysts such as SnCl₂ in governing the structure and biodegradability are expounded in detail. The degradation of PLA in SBF and its interaction with β-TCP particles constitute crucial aspects. For starch-based polymers, the enzymatic degradation catalyzed by amylase and glucosidase and the environmental impacts of temperature and humidity, in addition to the structural ramifications of amylose and amylopectin, are further elucidated. In plant fiber-based polymers, the biodegradation of cellulose and the effects of plasma treatment, electron beam irradiation, nanoparticles, and crosslinking agents on water resistance and stability are explicated with experimental substantiation. This manuscript also delineates technological accomplishments. PLA incorporated with HA demonstrates enhanced biocompatibility and finds utility in drug delivery systems. Starch-based polymers can be engineered for tailored degradation. Plant fiber-based polymers acquire water resistance and durability through specific treatments or the addition of nanoparticles, thereby widening their application spectrum. Synthetic and surface modification methodologies can be harnessed to optimize these materials. This paper also consolidates reaction conditions, research techniques, their merits, and demerits and delves into the biodegradation reaction mechanisms of these polymers. A comprehensive understanding of these degradation mechanisms is conducive to their application and progression in the context of sustainable development and environmental conservation. Full article

As major structural components of plant cell walls, cellulose and hemicellulose are degraded and fermented by anaerobic microbes in the rumen to produce volatile fatty acids, the main nutrient source for the host. Cellulose degradation is carried out primarily by specialist bacteria, with additional contributions from protists and fungi, via a variety of mechanisms. Hemicelluloses are hydrolyzed by cellulolytic bacteria and by generalist, non-cellulolytic microbes, largely via extracellular enzymes. Cellulose hydrolysis follows first-order kinetics and its rate is limited by available substrate surface area. Nevertheless, its rate is at least an order of magnitude more rapid than in anaerobic digesters, due to near-obligatory adherence of microbial cells to the cellulose surface, and a lack of downstream inhibitory effects; in the host animal, fiber degradation rate is also enhanced by the unique process of rumination. Cellulolytic and hemicellulolytic microbes exhibit intense competition and amensalism, but they also display mutualistic interactions with microbes at other trophic levels. Collectively, the fiber-degrading community of the rumen displays functional redundancy, partial niche overlap, and convergence of catabolic pathways that all contribute to stability of the ruminal fermentation. The superior hydrolytic and fermentative capabilities of ruminal fiber degraders make them promising candidates for several fermentation technologies. Full article

Application of thermoplastic fiber-reinforced lightweight composite materials provides a wide range of advantages that are of particular importance for the mobility sector. UD tapes composed of unidirectionally (UD) oriented inorganic fibers embedded in a thermoplastic matrix represent light-weight materials with high tensile strength. This publication addresses recycling aspects of novel UD tape made of a combination of basalt fibers and different PLA (polylactic acid) formulations. The kinetics of enzyme-based separation of polymer from the fiber were investigated. Different types of UD tapes with a thickness of 270–290 µm reinforced with basalt fiber weight ratios ranging between 51 and 63% were incubated at 37 °C in buffer solution (pH 7.4) containing proteinase K. The influence of enzyme concentration, tape weight per incubation tube, proteinase K activators, and tape types on the rate of enzymatic decomposition was investigated. Enzyme activity was measured by analyzing lactate concentration with lactate dehydrogenase and by measuring weight loss of the composite material. The rate of lactate release increased in the first 30 min of incubation and remained stable for at least 90 min. Weight loss of 4% within 4 h was achieved for a tape with 56% (w/w) fiber content. For a sample with a surface area of 3 cm² in a buffer volume of 10 mL, the rate of lactate release as a function of enzyme concentration reached saturation at 300 µg enzyme/mL. With this enzyme concentration, the rate of lactate release increased in a linear manner for tape surface areas between 1 and 5 cm². Four tapes with different PLA types were treated with the enzyme for 17 h. Weight loss ranged between 7 and 24%. Urea at a concentration of 0.5% (w/v) increased lactate release by a factor of 9. Pretreatment of tapes in alkaline medium before enzymatic degradation increased weight loss to 14% compared to 5% without pretreatment. It is concluded that enzymatic PLA hydrolysis from UD tapes is a promising technology for the release of basalt fibers after alkaline pretreatment or for the final cleaning of basalt fibers. Full article

The objectives of this study were to (1) determine the effect of exogenous fibrolytic enzyme derived from Trichoderma reesei on dry matter (DM) and neutral detergent fibre (NDF) degradability of whole plant faba bean silage (Snowbird), (2) evaluate the effects of fibrolytic enzyme (FETR) on DM and NDF degradation kinetics of whole plant faba bean silage, and (3) compare the difference between in the vitro approach (Daisy^II incubation method) and the in situ assay-biological approach (nylon bag technique) in the determination of degradability of dry matter (DMD) and neutral detergent fibre (NDFD). The fibrolytic enzyme from Trichoderma reesei was a mixture of xylanase and cellulase. The whole plant faba bean silage was treated with seven doses of fibrolytic enzyme, with 0 as a control and 0.25, 0.5, 0.75, 1, 1.25 and 1.5 mL of FETR/kg DM of silage. The results obtained from the in situ method show that fibrolytic enzyme cubically (p < 0.05) affected DMD and quadratically (p < 0.01) affected NDFD with increasing level of enzyme application. In vitro DM and NDF degradability were quadratically and cubically (p < 0.01) affected by the increasing dosage of enzyme. Correlation analysis between the in situ assay-biological approach and the In vitro Daisy^II approach showed a strong correlation (r = 0.98, p < 0.01) on overall DMD and also a satisfactory relationship (r = 0.84, p < 0.01) was found on overall NDFD. The enzyme application showed a great impact on NDF rumen degradation kinetics by decreasing the undegradable fraction and increasing the potential degradable fraction and the effective degradable content of fiber. The washable (W) and potential degradation fraction (D) were linearly (p = 0.05) increased by the enzyme treatments. Therefore, the undegradable fraction was linearly decreased (p = 0.05) with increasing dosage of enzyme. Both bypass (BNDF) and effective degradable NDF (EDNDF) were cubically (p = 0.05) affected by fibrolytic enzyme. In conclusion, the exogenous fibrolytic enzyme derived from Trichoderma reesei highly impacted rumen degradation characteristics and degradability of whole plant faba bean silage and could be used to improve fibre digestion of whole plant faba silage in dairy cows. Full article

Non-digestible polysaccharides are of great significance to human and animal intestinal health. Cellulose, arabinoxylan, β−glucan and glucomannan were selected in the present study to investigate the fermentation characteristics and fiber-degrading enzyme kinetics by inoculating pig fecal microbiota in vitro. Our results showed that fermentation of arabinoxylan and β-glucan produced the highest amount of acetate and lactate, respectively. The abundance of Prevotella_9 was the highest in β-glucan group and positively correlated with lactate and acetate. Glucomannan fermentation produced the highest amount of butyrate, and the abundance of Lachnospiraceae_XPB_1014_group and Bacteroides were the lowest. A significant negative correlation was found between Lachnospiraceae_XPB_1014_group, Bacteroides and butyrate. Exo-β-1,4-xylanase had the highest activity at 24 h during arabinoxylan fermentation. The activity of β-glucosidase and β-mannosidase at 36 h were higher than those at 15 h in the glucomannan group. The abundance of Prevotella_9 was positively correlated with β-glucosidase while Lachnospiraceae_XPB_1014_group and Bacteroides were negatively correlated with β-xylosidase. Our findings demonstrated the β-glucan and arabinoxylan promote proliferation of Prevotella_9, with the preference to secret β-glucosidase, β-mannosidase and the potential to produce lactate and acetate. Butyrate production can be improved by inhibiting the proliferation of Lachnospiraceae_XPB_1014_group and Bacteroides, which have the lack of potential to secret β-xylosidase. Full article