This study Aimed to evaluate the effects of Lactiplantibacillus plantarum (LP), cellulase (CE) or xylanase (XE) on the fermentation quality and microbial community of the broom sorghum stover silage. It was found that additives can improve the fermentation quality and digestibility of broom sorghum stover. Compared with the CK treatment, the addition of cellulase significantly increased the CP and WSC contents of broom sorghum stover, and also significantly improved thein vitrodry matter digestibility andin vitroneutral detergent fiber digestibility. The microbial community in broom sorghum stover silage also changed with different additive treatments. The microbiota shifted fromPantoeaandNissabactertoLactiplantibacillusandLentilactobacillus. Changes in microorganisms indicate a decrease in the number of harmful microorganisms and more retention of nutrients. All treatments reduced pungent VOCs (e.g., Benzyl isothiocyanate) and enriched aromatic VOCs (e.g., Phenol 4-ethyl-, β-damascenone), with the CE treatment performing best. Ensiling degraded anti-nutritional factors: the CK and LP treatments were superior for tannin reduction, the CE and XE treatments for phytic acid. These additives effectively enhance broom sorghum stover's feed value, with CE showing the most comprehensive effect, providing a basis for its high-value utilization and sustainable livestock production.

The precise organization of multienzymes on biomaterial surfaces remains a fundamental challenge in designing high-performance biohybrid systems. In this work, a bioinspired, hypothesis-driven platform, multitemplate artificial antibody-antigen directed immobilization (MT-3ADI), is introduced that enables programmable modular assembly of multiple enzymes. This "plug-and-play" architecture dictates structure-function relationships while facilitating enzyme exchange and system regeneration. Applied to purpurogallin synthesis, the engineered glucose oxidase (GOx)-horseradish peroxidase (HRP) and cellulase (CEL)-GOx-HRP cascades achieved exceptional yields (96.31 ± 0.13% and 90.25 ± 0.21%) and robust operational stability (retaining >80% activity after six cycles). By establishing a generalizable framework for controlled protein organization, this work addresses core priorities in biomaterial surface science and offers broad applicability from green synthesis to therapeutic delivery.

PurposeDespite the importance of gerbera in the international market, stem bending is still the biggest problem for gerbera cut flowers during the post-harvest stage, as it reduces the flowers’ vase life and their market value. MethodsTo address this problem, the effects of different vase solutions containing calcium chloride (CaCl2) and sodium selenite (Na2SeO3) on the longevity and stem curvature of gerbera ‘Stanza’ flowers were investigated using repeated-measures ANOVA. The treatments were: distilled water (control, T1), 1 g L–1 (T2) and 2 g L–1 (T3) of CaCl2, 1 mg L–1 (T4) and 2 mg L–1 (T5) of Na2SeO3, and 1.5 g L–1 CaCl2 plus 1.5 mg L–1 Na2SeO3 (T6). Results Gerbera cut flowers subjected to T6 of vase solution exhibited a significantly longer vase life (20 days) and a substantially lower degree of stem bending (25.7°). Moreover, these flowers achieved the lowest significant ion leakage (IL) in petals, and an inhibition in the cellulase (CEL), polygalacturonase (PG), xylanase (XYL), and pectinase (PT) activities, along with noticeably greater levels of catalase (CAT) activity and lignin content until the last day of sampling. Automatic linear modeling analysis revealed CEL, PT, PG, lignin content, and XYL were selected as the most contributing attributes in determining the degree of stem bending. Conclusions It can be recommended that using calcium and/or selenium can be regarded as an effective substitute vase solution to maintain gerbera flowers and reduce bending stems over the post-harvest period.

ABSTRACT Conservation agriculture (CA)-based practices reduce the risk of soil nutrient depletion by promoting the stabilization and sequestration of organic carbon (C) and enhancing soil biological functioning. To address this, a field study was conducted to quantify the impact of rice establishment methods, tillage and residue management practices for eight-years to identify the critical growth stages of rice and wheat for assessing the biological properties of soil under rice-wheat cropping system (RWS). The results revealed that direct seeded rice-reduced tillage followed by zero tillage (ZT) with residue retention significantly improved soil's extracellular enzyme activities except phenol oxidase (PHEOX) and peroxidase (PERO) across five growth stages of rice-wheat systems. Dehydrogenase (DHA), fluorescein diacetate (FDA), alkaline phosphatase (Alk-P), β-glucosidase (β-glu), total polysaccharides carbon (TPC) and cellulase (CELL) activities were highest at the maximum tillering stage of rice and wheat than at other growth stages. At the maximum tillering stage of wheat, activities of DHA, FDA, Alk-P, β-glu and TPC under ZT-wheat with residue retention were 60.5, 35.8, 50.6, 30.4, and 31.1% higher than conventional tillage (CT) without residue. Furthermore, ZT-wheat with residue retention significantly increased wheat grain yield by 10.6% as compared to CT without residue. Principal component analysis (PCA) suggested that xylanase, phenol oxidase and FDA can be used as sensitive parameters for assessing soil quality under rice establishment methods, tillage and residue management practices in RWS. These findings highlight that practices like ZT with residue retention support sustainable intensification of RWS by enhancing soil biological fertility and crop productivity.

Fungal enzymes in glycoside hydrolase family 5 subfamily 5 (GH5_5) display notable catalytic diversity, efficiently degrading cellulose and sometimes mannan. However, the structural determinants and molecular mechanisms governing substrate preference in this enzyme family remain unclear. In this study, GH5_5 enzymes from fungi were systematically classified using profile-based sequence models and functionally characterized. Saturation mutagenesis combined with high-resolution crystal structure analysis of the bifunctional enzyme BsCel5B, exhibiting cellulase (CEL) activity of 941 ± 17 U/mg and mannanase (MAN) activity of 1,736 ± 34 U/mg, was employed to identify key residues controlling substrate specificity. Residue T100 in BsCel5B was identified as a major structural contributor associated with significant shifts in substrate preference. The T100V and T100N mutations resulted in 2.0-fold increases in MAN activity and 2.5-fold increases in CEL activity, respectively, generating bifunctional enzymes with enhanced substrate-specific activities. Similar substrate specificity trends were observed in several GH5_5 cellulase mutants. Their structural analysis indicated that substrate preference in fungal GH5_5 enzymes might be shaped by residual network-mediated alterations of the active-site geometry, with T100 acting as a second-shell regulatory element within a cooperative residue network. Together, these findings suggest a mechanistic framework for engineering catalytic specificity in GH5_5 enzymes.IMPORTANCECellulose and mannan are major components of plant biomass, and enzymes capable of efficiently breaking them down are essential for sustainable biofuel production and biomass utilization. Fungal enzymes in GH5_5 are widely used for these purposes, yet their functional diversity has been difficult to predict or control. Substrate preference in these enzymes can be modulated by altering a single amino acid, offering a promising approach for tuning enzyme activity. The identification of a key residue that influences the balance between cellulose and mannan degradation provides valuable insights for engineering enzymes with tailored functions. These findings contribute to a deeper understanding of fungal biomass-degrading enzymes and support the rational design of more efficient catalysts for industrial and environmental applications.

Using cellulase to degrade cellulose for cellobiose production is an economically competitive pathway for the high-value utilization of lignocellulose. Cellobiohydrolases hydrolyze cellulose into cellobiose but suffer severe product inhibition, whereas endoglucanases (EGs) randomly cleave cellulose chains into oligosaccharides. Three heterologously expressed EGs from different sources (PcCel45A, BsCel5, and GtCel5A) were screened for their ability to degrade cellulose into cellobiose, and their hydrolysis processes were characterized. Using PcCel45A and BsCel5 to hydrolyze Avicel step-by-step at a protein ratio of 1:1 produced cellobiose with a purity of 90.99% and a yield of 1.2 g per 100 g of Avicel. Further studies revealed that PcCel45A and BsCel5 cannot degrade cellobiose, and their activities are not inhibited by the product, showing their advantage in cellobiose production. Additionally, adsorption kinetics and thermodynamic analyses indicated that PcCel45A and BsCel5 exhibited distinct adsorption mechanisms. This study provides a promising new strategy for the high-value utilization of lignocellulose.

Abstract In this study, a constrained mixture design was applied to evaluate the potential of agro-industrial by-products, namely cocoa fruit shell (CFS), cocoa bean shell (CBS), and palm oil (PO) as substrates for enzyme production. A desirability function approach was employed to optimize the simultaneous production of endoglucanase (EGL), exoglucanase (EXG), and β-glucosidase (BGL) by Penicillium roqueforti ATCC 10110 under solid-state fermentation (SSF). The desirability function reached a value of 0.84, indicating successful fulfillment of the optimization criteria. The optimized substrate proportions CFS (7.5 g), CBS (1.0 g), and PO (1.5 g) resulted in enzyme activities of (5.38 IUg −1 ) for EGL, (1.01 IUg −1 ) for EXG, and (2358.75 IUg −1 for BGL). All enzymes exhibited high stability at 50 °C and at pH 4.5 and 6.0. The presence of CoSO 4 and EDTA enhanced the activities of EGL, EXG, and BGL by more than 100%. The effects of solvent addition were further analyzed using artificial neural networks based on a Kohonen Self-Organizing Map (KSOM) to identify correlations among the observed responses. Overall, the application of desirability based optimization combined with constrained mixture design proved effective in enhancing cellulase production by P. roqueforti ATCC 10110. The characterized enzymes demonstrated have shown significant potential for industrial applications. Graphical Abstract

In recent years, there has been increasing emphasis on the sustainable organic material reutilization, including straw and biochar. This study investigates the effects of prolonged applications of straw (R), biochar (B), and their combination (RB) on soil organic carbon fractions, enzyme activities, and tomato yield under continuous greenhouse cultivation, compared to a control (CK). The results showed that organic amendments reduced soil bulk density. Compared to the CK treatment, the R treatment caused a slight decrease in soil pH, whereas the B and RB treatments significantly increased soil pH by 4.10% and 6.10%, respectively. Relative to the CK treatment, the R, B, and RB treatments significantly elevated the levels of dissolved organic carbon (DOC), easily oxidizable organic carbon (ROC), microbial biomass carbon (MBC), and particulate organic carbon (POC) by ranges of 10.28%-24.34%, 38.94%-49.69%, 109.73%-121.21%, and 102.56%-196.93%, respectively. The activities of catalase (CAT), α-glucosidase (AG), β-glucosidase (BG), β-xylosidase (XYL) and cellulase (CL) were significantly enhanced after organic material addition. Additionally, the R, B, and RB treatments increased yields by 22.61%, 18.25%, and 23.74%, respectively. By integrating long-term field observations with structural equation modeling, this study offers novel mechanistic evidence that particulate organic carbon (POC) serves as a central mediator linking organic amendments to yield improvement-a regulatory pathway that has not been systematically elucidated in previous research, offering a mechanistic basis for sustainable soil management in continuous greenhouse production.

The aim of this study was to evaluate the effects of enzymes and lactic acid bacteria (LAB) on the fermentation characteristics, bacterial community and predicted functional profiles of whole hulless barley. The hulless barley was inoculated with distilled water, Lactobacillus plantarum, L. plantarum and cellulase (CE), L. plantarum and L. buchneri, as the CON, T1, T2 and T3 treatments, respectively. After 60 days of fermentation, the T3 treatment had the lowest (p < 0.05) pH and highest (p < 0.05) lactic acid (LA) content. Additionally, a significantly (p < 0.05) higher content of propionic acid (PA) was found in the CON and T3 treatments compared to the T1 treatment. At the phylum level, Firmicutes was the dominant phylum in all treatments. At the genus level, Lactiplantibacilus was the dominant genus in all treatments. The relative abundance of Lentilactobacillus was higher and Pantoea was lower in the T1 and T3 treatments. Functional prediction analyses showed that T3 treatment had lower carbohydrate metabolism and higher cofactor and vitamin metabolism. In summary, the inoculation of Lactobacillus and cellulase modulated the bacterial community to improve the silage quality of hulless barley, and the addition of complex LAB show better results.

Antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and virulence factors (VFs) represent growing threats to global public health. Sophora davidii silage is a common feed in livestock production, may potentially serve as a reservoir and vector for the dissemination of these genetic determinants. The biosafety risks associated with Sophora davidii silage remain poorly evaluated. Consequently, mitigating these risks through improved processing techniques has become an urgent priority. This study systematically elucidates the effects of additives and chopping lengths on the microbial community structure, ARGs, MGEs, and VFs during the ensiling of Sophora davidii. Metagenomic analyses demonstrated that both additives and chopping length significantly influenced the biosafety profile of Sophora davidii silage. Additives markedly reduced the abundance of ARGs, MGEs, and VFs (P < 0.05). Formic acid (FA) demonstrated the most pronounced suppression, whereas cellulase (CE) was least effective. Notably, although inoculation with Lactiplantibacillus plantarum (LP) reduced the overall abundance of these risk factors, the strain may be associated with the vanY gene in the vanB cluster, mobile genetic elements (MGEs) such as ISLpl1 and ISLp1, as well as specific virulence factors (VFs). Furthermore, at a 5cm chopping length, the Control group exhibited significantly higher levels of ARGs and VFs compared to the 1cm and 3cm treatments (P < 0.05). The findings highlight the efficacy of reducing chopping length (to 1cm) in controlling the proliferation and dissemination of ARGs, MGEs, and VFs. Moreover, the use of direct acidifying agents, particularly formic acid, offers considerable advantages in enhancing the microbial safety of silage.

Ferulic acid esterase (FAE) catalyzes the hydrolysis of the feruloyl ester bond in lignocellulose, exposing cellulose. The objective of this research was to examine the impacts of Bacillus amyloliquefaciens A30 producing FAE on the fermentation quality, fiber degradation, enzyme activity and microbial diversity of corn bran silage and whole-plant corn silage. The experimental treatments were as follows: control (CK), cellulase (CEL), strain A30 (A30) and CEL + A30. Corn bran and whole-plant corn were ensiled for 14 d and 60 d, respectively. The results showed that all additive treatments effectively reduced the pH, neutral detergent fiber, acid detergent fiber and cellulose contents of both corn bran silage and whole-plant corn silage in comparison with control, with CEL + A30 group performing the best effects. Meanwhile, higher FAE activity was detected in A30 and CEL + A30 groups during ensiling. Furthermore, the supplementation of A30 increased the degradation ratio of NDF, ADF, ADL, and cellulose of corn bran silage and whole-plant corn silage. Additionally, treatments with A30 and CEL + A30 increased the abundance of Lactobacillus, and reduced the proportion of pathogenic genera, including Acinetobacter, Enterobacter, and Sphingobacterium. In conclusion, the application of A30 may effectively promote fiber degradation and the stability of microecological system for corn silage.

Laboratory incubation experiments were conducted to investigate the effects of coal fly ash (FA) amendment (0%, 2.5%, 7.5%, and 15%) and moisture regimes (40%, 70%, and 100% water holding capacity (WHC)) on the mineralization of carbon (C) in an acidic agricultural soil. The results showed that the soil C mineralization intensity initially increased and subsequently decreased throughout the incubation period, with the mineralization dynamics well described by the first-order kinetic model (0.9633 ≤ R2 ≤ 0.9972). Carbon mineralization increased with the application rate of FA, while moisture effect followed the order 70% WHC &gt; 100% WHC &gt; 40% WHC. Indicators showing highly significant correlations with total C mineralization amount included FA application rate, pH, water-soluble organic carbon, (WSOC) and cellulase (CEL) activity. Specific bacterial (Acidobacteriota, Gemmatimonadota, Pseudomonadota, and Actinobacteriota) and fungal phyla (Chytridiomycota, Glomeromycota, and Olpidiomycota) exhibited stronger correlations with C mineralization. The microbial taxa exhibiting significant responses to FA and moisture conditions were not consistent. Although the addition of high proportions of FA, especially with adequate moisture conditions, can enhance soil microbial activity and C mineralization, the potential risks of soil C loss and the accumulation of toxic elements necessitate the prudent implementation of elevated FA application rates in practical scenarios.

Abstract Ensiling tropical grasses presents a challenge in achieving an optimal fermentation profile, particularly when the grass possesses high nutritional value. Therefore, this study aimed to evaluate the use of additives during ensiling as a strategy to enhance the fermentation profile and nutritional quality of Zuri grass ( Megathyrsus maximus cv. BRS Zuri) silage harvested at two regrowth ages. The tested additives were control (CT); Lactiplantibacillus pentosus strain AV 14.17 (LP); cellulase (CE); and LPCE (LP + CE). The regrowth ages were 60 and 90 days. Increased regrowth age of Zuri grass resulted in a higher fermentability coefficient but a reduced crude protein content in the fresh forage. The CE and LPCE silages exhibited lower fibre content. The crude protein content was higher and the ammonia content was lower in the CE and LPCE silages when the grass was harvested at 60 days. The pH was lower in CE and LPCE silages at both regrowth ages. Butyric acid was detected only in the CT and LP silages when the grass was harvested at 60 days. The CE and LPCE silages showed greater effective digestibility. The indigestible fraction of neutral detergent fibre was higher in silage harvested at 90 days. The addition of cellulase, alone or combined with the L. pentosus strain, effectively solubilised the fibre components, resulting in a lower pH and more effective control of undesirable microorganisms, thus improving the nutritional value. The most significant results were obtained when Zuri grass was harvested at 60 days of regrowth.

The objective of this study was to explore the characteristics of soil organic carbon （SOC） mineralization of latosol subjected to continuous application of chemical fertilizers in combination with various organic materials. Soils were sampled from a typical pepper-corn rotation system in Hainan Province， where five treatments were established five years ago： a control （CK）， chemical fertilizer alone （NPK）， chemical fertilizer combined with corn straw （NPK+S）， chemical fertilizer combined with biochar （NPK+B）， and chemical fertilizer combined with sheep manure （NPK+M）. Then， a controlled indoor mineralization experiment was conducted. The driving factors of SOC mineralization were analyzed by measuring carbon conversion-related enzyme activities and the 13C nuclear magnetic resonance method. The results revealed that the combined application of organic materials significantly increased the contents of SOC， dissolved organic carbon （DOC）， and microbial biomass carbon （MBC） by 3.32%-75.08%， 219.32%-477.30%， and 5.12%-52.78% compared to the NPK treatment， respectively. Among these treatments， NPK+B had the greatest impact on enhancing SOC content， while NPK+M significantly increased MBC and DOC levels. Furthermore， NPK combined with organic materials significantly enhanced cumulative SOC mineralization by 86.83%-280.94%， following the order NPK+M&gt;NPK+S&gt;NPK+B. Compared with the NPK treatment， NPK+B treatment significantly increased the ratio of hydrophobic carbon to hydrophilic carbon and aromaticity by 48.31% and 105.89%， respectively. The NPK+M treatment significantly increased soil β-glucosidase （BG）， cellulase （CL）， and sucrase （SC） activities （P&lt;0.05）， whereas NPK+S notably enhanced CL and SC enzyme activities （P&lt;0.05）. By contrast， the NPK+B treatment exhibited relatively lower BG， POD， and SC enzyme activities. The structure equation model showed that soil active carbon components and carbon conversion-related enzyme activity directly regulated SOC mineralization. Both redundancy analysis and correlation analysis further revealed that cumulative SOC mineralization was strongly positively correlated with the contents of MBC and DOC and the activities of BG and SC. Consequently， the combination of chemical fertilizer with biochar appears to be the most effective approach for increasing the SOC content of latosol by enhancing recalcitrant carbon components and reducing carbon-converting enzyme activities， thereby resulting in relative lower organic carbon mineralization. The results of this study provide valuable scientific insights for optimizing fertilization strategies and enhancing carbon sequestration potential in tropical farmlands.

Ferulic acid possesses certain antioxidant and antibacterial properties. Additionally, ferulic acid esterase (FAE) and cellulolytic enzymes have been associated with synergistic degradation of ferulic acid ester bonds, thereby facilitating greater release of ferulic acid from lignocellulose, which could have important effects on silage quality and aerobic stability. This study examined the effects of ensiling Broussonetia papyrifera with FAE-producing Lactiplantibacillus plantarum (LP), cellulase (CE) and xylanase (XY) under aerobic exposure conditions. The following treatments were used: distilled water (CK), LP, LP + CE, LP + XY and LP + XY + CE. After 60 days of silage treatment, the samples were unsealed for aerobic exposure for 1, 3, 5, or 7 days. Compared with the CK treatment, the addition of FAE-producing L. plantarum significantly (P < 0.05) led to lower pH, reduced dry matter loss of the silage and increased lactic acid (LA) concentration after 60 d of ensiling (especially for the LP + CE and LP + CE + XY groups). During the aerobic exposure stage, the combined treatment with LP and enzymes effectively inhibited the increase in pH, significantly reduced the rate of dry matter loss and increased the LA concentration and aerobic stability of the silage (P < 0.05). Moreover, the LP + CE and LP + CE + XY treatment groups exhibited higher ferulic acid levels than the other groups did, corresponding with greater aerobic stability, especially for the LP + CE group, which remained stable. In this group, the pH values showed minimal change, increasing by only 0.31 (4.24-4.55) after 7 days of aerobic exposure. In addition, the LP and enzyme co-treatment was linked to shifts in the microbial community of the silage during aerobic exposure, with increased relative abundance of Lactiplantibacillus plantarum, and its abundance positively correlated with lactic acid and ferulic acid concentrations, while negatively correlated with ammonia nitrogen; and inhibited proliferation of spoilage-related bacteria (Enterobacter, Gluconobacter and Cladosporium). The combination of FAE-producing L. plantarum and cellulase can be used as an effective method to increase the preservation efficiency and aerobic stability of B. papyrifera silage.

Cotton Verticillium wilt is a disease that significantly impacts the cotton industry, severely affecting cotton quality and the economic well-being of farmers. Bacillus atrophaeus YL84 is a biocontrol bacterium with broad-spectrum antagonistic and growth-promoting characteristics, previously isolated by our laboratory. This study aimed to elucidate the antagonistic effects of sterilized fermentation filtrate from Bacillus atrophaeus YL84 on cotton Verticillium wilt pathogen Verticillium dahliae and its growth-promoting effects on cotton. The experiments were conducted in vitro and in vivo to assess these effects comprehensively. Using the dual culture method, it was found that Bacillus atrophaeus YL84 exhibited a high inhibition rate on mycelial growth of V. dahliae, with an inhibition rate of 84.11%. The undiluted YL84 sterilized fermentation filtrate and its 10% volume fraction dilution (fermentation filtrate diluted to 10%) exhibited inhibition rates of 80.25% and 72.16% for conidial germination and mycelial growth of V. dahliae, respectively. Scanning electron microscopy showed increased branching, swelling, and shortened internodes in the antagonized mycelia. Conductivity measurements revealed a significant enhancement caused by the YL84 filtrate, with conductivity increasing by 8.94 times compared to the control at a 250 μg/mL concentration. Similarly, protein leakage peaked at 9.47 times the control level at 250 μg/mL, demonstrating the filtrate’s potent impact on mycelial cell membrane permeability. The enzymatic activities of polygalacturonase (PG), cellulase (CL), and β-glucosidase (β-GC) were significantly reduced following treatment with YL84 sterilized fermentation filtrate, with reductions from control levels of 15.78, 10.11, and 5.01 U/mL to treatment levels of 11.81, 6.96, and 1.44 U/mL, respectively. Indoor pot experiments demonstrated that different concentrations of YL84 sterilized fermentation filtrate significantly suppressed the occurrence of cotton Verticillium wilt while promoting plant growth. Compared to the control group, application of 250 μg/mL YL84 sterilized fermentation filtrate resulted in a control efficacy of 66.69% for cotton Verticillium wilt, with increases in plant height, root length, fresh weight, and dry weight of 9.36–33.85%, 17.33–29.49%, 16.79–28.24%, and 25–58.33%, respectively. These findings underscore the potential of the YL84 filtrate as both a biocontrol agent and a promoter of cotton plant growth in agricultural settings. These results indicate that Bacillus atrophaeus YL84 sterilized fermentation filtrate possesses both disease-suppressing and growth-promoting activities, making it a promising candidate for development and use as a biocontrol agent and plant growth promoter.

Abstract Persimmon is a climacteric fruit that is highly prone to chilling injury (CI) and softening, which limits its long-term marketability. This study investigated the effectiveness of treatments on persimmon fruit: a 5 mM oxalic acid (OA) dip treatment for 2 min, modified atmosphere packaging (MAP), their combination (OA + MAP), and untreated fruits were considered as control, followed by storage at 0 ± 1 °C for up to 60 days with subsequent 1 day at shelf conditions. The combined OA + MAP treatment alleviated CI incidence, followed by MAP alone and OA treatment in persimmon fruit. Additionally, the OA + MAP treatment significantly suppressed the ethylene production rate (EPR), minimised weight loss (WL), and maintained better fruit texture, including hardness and chewiness. The treatment also effectively reduced malondialdehyde (MDA) production and the activity of lipoxygenase (LOX), as well as hydrogen peroxide (H 2 O 2 ) production in persimmons. Analysis of cell wall components revealed that OA + MAP-treated fruit maintained higher levels of total pectin (TP), chelate soluble pectin (CSP), sodium carbonate soluble pectin (NSP), and cellulose. Furthermore, separate application of OA or MAP and the combined OA + MAP treatment delayed the activities of pectin methyl esterase (PME), pectate lyase (PL), polygalacturonase (PG), and cellulase (CEL) in comparison with control fruit during storage. These findings demonstrate that the combined application of OA + MAP effectively enhances chilling tolerance and preserves cell wall integrity in persimmons.

ABSTRACT This study was performed to evaluate the impact of fibrolytic enzymes on the chemical composition, fermentation profile, microbial population, and in vitro dry matter (DM) and neutral detergent fibre (NDF) degradability of silage from two Urochloa species. The experiment followed a completely randomised 4 (fibrolytic enzymes) × 2 (species) factorial design with three replicates. The fibrolytic enzymes were: control (CT), cellulase (CE) (0.1% fresh matter), xylanase (XL) (400 mg/kg of DM), and a blend (CEXL) (cellulase + xylanase). The species tested were U. decumbens cv. Basilisk and U. brizantha cv. BRS Piatã. The CE treatment resulted in a lower pH and a greater lactic acid bacteria population. The CEXL treatment increased crude protein content, while CE and CEXL reduced ammonia, NDF, acid detergent fibre, and lignin levels compared with CT. The XL treatment increased the potentially soluble fraction and the DM degradation rate, while CE and the blend increased the soluble fraction of DM. Silage treated with CEXL had lower indigestible NDF, while CE increased the degradation rate. The U. brizantha exhibited a greater NDF degradability and degradation rate than U. decumbens. Therefore, CE is recommended to improve the chemical composition, fermentation profile, and degradability of Urochloa silages.

Toona sinensis (T. sinensis) is a popular woody vegetable with distinct red and green varieties in China. Despite its significance, research on the comparative nutrient composition profiles and cellulose and hemicellulose dynamics between these two varieties remains limited. This study comprehensively investigated the refrigerated storage characteristics of T. sinensis buds from multiple aspects, related to cellulose and hemicellulose synthesis. The results showed marked differences between the two varieties during 3 d postharvest storage. Green T. sinensis buds had more severe blackening at the petiole base. Green T. sinensis buds were also richer in vitamin C (Vc), protein, reducing sugars, flavonoids, and total phenols, while red T. sinensis buds had higher total sugar content. In terms of enzyme activities, red T. sinensis buds had elevated β-xylosidase metabolizes hemicellulose content over 28.65 mg·g-1 higher than that of green T. sinensis buds, while green T. sinensis buds increased cellulase (CL) activity led to a hemicellulose content 26.60 mg·g-1 high than red T. sinensis buds. The cell wall thickening and polygonal cell shape during storage were closely associated with the increase in hemicellulose content. Additionally, red T. sinensis buds exhibited elevated CAT and SOD activities in response to oxidative stress induced by increased MDA levels. In summary, green T. sinensis buds demonstrated higher nutritional value but reduced storage stability and enhanced lignification compared to red T. sinensis buds. This research not only provides a multi-dimensional understanding of T. sinensis storage characteristics, but also lays a foundation for the development of scientific storage and preservation methods.

Ultraviolet (UV)-induced mutagenesis is a cost-effective and straightforward technique for introducing random genetic variations without the use of chemical reagents or genetic engineering. It is commonly employed to enhance enzyme activity in industrial trains. In this study, Trichoderma sp. was exposed to UV radiation at varying distances (4, 9, and 13 cm) and durations (2, 4, 6, and 8 min) to induce mutations. The activities of endoglucanase (EG), β-glucosidase (BGL), and cellobiohydrolase (CBH) were assessed following treatment. The 4 cm exposure distance yielded the highest enhancement, with EG, BGL, and CBH activities increasing 1.5-, 1.3-, and 0.9-fold, respectively. When the distance was fixed at 4 cm, the optimal exposure time was identified as 4 min, yielding further enhancements of 1.9-, 1.6-, and 1.4-fold, respectively. The resulting mutant, designated Mut-4, was scaled up in a 10-L bioreactor to assess its industrial applicability. Mut-4 retained its enhanced performance, achieving 1.9-, 2.0-, and 1.4-fold enhancements in EG, BGL, and CBH activities, respectively, compared with the original strain. These findings indicate that combining UV-induced mutagenesis with basic screening is an effective strategy for enhancing cellulolytic enzyme production, representing a promising approach for lignocellulosic biomass conversion.