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The use of biochar has emerged a potentially effective approach to improve soil function and promote crop performance. However, the specific impact of biochar on ß-glucosidase enzyme activity (BGA) within crop rotation systems in the Black Sea agroecosystem requires further investigation. This study was conducted to determine the effects of rice husk biochar (RHB) and poultry manure biochar (PMB) on BGA in soils. Six biochar doses (0-control, 10, 20, 30, 40 and 50 t ha-1) were applied at the beginning of two wheat-cabbage red pepper rotation periods. The mean BGA at second rotation (73.71 µg pNP g-1) was significantly lower compared to the BGA of the first period (93.39 µg pNP g-1). The BGA value in control (94.51 µg pNP g-1) decreased with increasing biochar application doses (76.05 µg pNP g-1, 50 t ha-1) treatment. The mean BGA value in PMB treatment was slightly higher than that of RHB, but it was not statistically different between two biochar types. However, the decrease in BGA value (25.0%) in the highest RHB dose compared to control was more than two-fold compared to the decrease in PMB application (12.1%). The difference in carbon/nitrogen ratio between RHB and PMB can be attributed to the variation in BGA values observed at the application of same biochar doses. The decrease in BGA over the course of the two rotation cycles implies that biochar may have a long-term influence on soil carbon cycling. [ABSTRACT FROM AUTHOR]

Trichoderma reesei RUT-C30 was cultivated on differentially pretreated rice straw and pure cellulose as a carbon source/inducer for cellulase production, and the enzymes were evaluated for hydrolysis of sequential acid and alkali pretreated rice straw. Growth on pretreated rice straw enhanced protein secretion and cellulase activities compared to pure cellulose as a carbon source. The yield of cellulolytic enzymes was higher for alkali pretreated rice straw (ALP-RS), while H2O2-treated (HP-RS) could not induce cellulases to a larger level compared to pure cellulose. Protein concentration was 3.5-fold higher on ALP-RS as compared to pure cellulose, with a maximum filter-paper cellulase (FPase) activity of 1.76 IU/ml and carboxy-methyl cellulase (CMCase) activity of 40.16 IU/ml (2.18 fold higher). Beta-glucosidase (BGL) activity was more or less the same with the different substrates and supplementation of heterologous BGL could result in a quantum jump in hydrolytic efficiencies, which in the case of ALP-RS induced enzymes was 34% (increased from 69.26% to 92.51%). The use of lignocellulosic biomass (LCB) itself as a substrate for the production of cellulase is advantageous not only in terms of raw material costs but also for obtaining a more suitable enzyme profile for biomass hydrolysis. [ABSTRACT FROM AUTHOR]

Abstract Beta-glucosidases catalyze the hydrolysis of the glycosidic bonds of cellobiose, producing glucose, which is a rate-limiting step in cellulose biomass degradation. In industrial processes, β-glucosidases that are tolerant to glucose and stable under harsh industrial reaction conditions are required for efficient cellulose hydrolysis. In this study, we report the molecular cloning, Escherichia coli expression, and functional characterization of a β-glucosidase from the gene, CelGH3_f17, identified from metagenomics libraries of an Ethiopian soda lake. The CelGH3_f17 gene sequence contains a glycoside hydrolase family 3 catalytic domain (GH3). The heterologous expressed and purified enzyme exhibited optimal activity at 50 °C and pH 8.5. In addition, supplementation of 1 M salt and 300 mM glucose enhanced the β-glucosidase activity. Most of the metal ions and organic solvents tested did not affect the β-glucosidase activity. However, Cu2+ and Mn2+ ions, Mercaptoethanol and Triton X-100 reduce the activity of the enzyme. The studied β-glucosidase enzyme has multiple industrially desirable properties including thermostability, and alkaline, salt, and glucose tolerance.

The Aburrá-Medellín river has been affected by urbanization processes in its basin, which has altered its physicochemical profile. In order to identify patterns in the enzymatic activity associated with variations in water quality and flow regime, the activity of the ß-glucosidase and phosphatase enzymes, related to carbon and phosphorus metabolism, respectively, was measured in the water and biofilm. For this purpose, nine monitoring stations with different degrees of anthropic intervention were selected, which were evaluated during low, medium and high flow regimes associated with meteorological variability in the basin. The hypothesis was that the physicochemical profile, water quality and flow regime affect the activity of both enzymes, both in the water and in the biofilm, since they influence the concentration of nutrients transported by the river. According to the results obtained, no statistically significant differences were found in the enzymatic activity at low, medium and high flow rates, although a higher concentration of nutrients was observed at low and medium flow rates than at high flow rates. On the other hand, it was observed that the concentration of nutrients in the river changed according to the degree of urbanization in the basin, due to the discharge of wastewater from densely populated municipalities. The activity of both enzymes was higher in the stations with higher concentrations of nutrients, while in the monitoring stations located in the upper zone of the basin, where anthropic intervention and the concentration of nutrients was low, the lowest enzymatic activities were found both in the water and in the biofilm. In this regard, enzymatic activity can be used as an indicator of the health of the Aburrá-Medellín river and it is suggested that the measurement of this variable be included in the framework of the RedRío monitoring network. [ABSTRACT FROM AUTHOR]

The food enzyme β‐glucosidase (β‐D‐glucoside glucohydrolase; EC 3.2.1.21) is produced with the non‐genetically modified Penicillium guanacastense strain AE‐GLY by Amano Enzyme Inc. The food enzyme is intended to be used in four food manufacturing processes. Dietary exposure to the food enzyme‐total organic solids (TOS) was estimated to be up to 4.054 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90‐day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 943 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 233. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that the risk of allergic reactions by dietary exposure cannot be excluded, but the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use. [ABSTRACT FROM AUTHOR]

Trichoderma erinaceum is a filamentous fungus that was isolated from decaying sugarcane straw at a Brazilian ethanol biorefinery. This fungus shows potential as a source of plant cell wall-degrading enzymes (PCWDEs). In this study, we conducted a comprehensive multiomics investigation of T. erinaceum to gain insights into its enzymatic capabilities and genetic makeup. Firstly, we performed genome sequencing and assembly, which resulted in the identification of 10,942 genes in the T. erinaceum genome. We then conducted transcriptomics and secretome analyses to map the gene expression patterns and identify the enzymes produced by T. erinaceum in the presence of different substrates such as glucose, microcrystalline cellulose, pretreated sugarcane straw, and pretreated energy cane bagasse. Our analyses revealed that T. erinaceum highly expresses genes directly related to lignocellulose degradation when grown on pretreated energy cane and sugarcane substrates. Furthermore, our secretome analysis identified 35 carbohydrate-active enzymes, primarily PCWDEs. To further explore the enzymatic capabilities of T. erinaceum, we selected a β-glucosidase from the secretome data for recombinant production in a fungal strain. The recombinant enzyme demonstrated superior performance in degrading cellobiose and laminaribiose compared to a well-known enzyme derived from Trichoderma reesei. Overall, this comprehensive study provides valuable insights into both the genetic patterns of T. erinaceum and its potential for lignocellulose degradation and enzyme production. The obtained genomic data can serve as an important resource for future genetic engineering efforts aimed at optimizing enzyme production from this fungus. [ABSTRACT FROM AUTHOR]

Beta-glucosidases catalyze the hydrolysis of the glycosidic bonds of cellobiose, producing glucose, which is a rate-limiting step in cellulose biomass degradation. In industrial processes, β-glucosidases that are tolerant to glucose and stable under harsh industrial reaction conditions are required for efficient cellulose hydrolysis. In this study, we report the molecular cloning, Escherichia coli expression, and functional characterization of a β-glucosidase from the gene, CelGH3_f17, identified from metagenomics libraries of an Ethiopian soda lake. The CelGH3_f17 gene sequence contains a glycoside hydrolase family 3 catalytic domain (GH3). The heterologous expressed and purified enzyme exhibited optimal activity at 50 °C and pH 8.5. In addition, supplementation of 1 M salt and 300 mM glucose enhanced the β-glucosidase activity. Most of the metal ions and organic solvents tested did not affect the β-glucosidase activity. However, Cu2+ and Mn2+ ions, Mercaptoethanol and Triton X-100 reduce the activity of the enzyme. The studied β-glucosidase enzyme has multiple industrially desirable properties including thermostability, and alkaline, salt, and glucose tolerance. [ABSTRACT FROM AUTHOR]

In the present study, a thermophilic strain designated CamBx3 was isolated from the Campanario hot spring, Chile. Based on 16S rRNA gene sequence, phylogenomic, and average nucleotide identity analysis the strain CamBx3 was identified as Bacillus paralicheniformis. Genome analysis of B. paralicheniformis CamBx3 revealed the presence of genes related to heat tolerance, exopolysaccharides (EPS), dissimilatory nitrate reduction, and assimilatory sulfate reduction. The pangenome analysis of strain CamBx3 with eight Bacillus spp. resulted in 26,562 gene clusters, 7,002 shell genes, and 19,484 cloud genes. The EPS produced by B. paralicheniformis CamBx3 was extracted, partially purified, and evaluated for its functional activities. B. paralicheniformis CamBx3 EPS with concentration 5 mg mL-1 showed an optimum 92 mM ferrous equivalent FRAP activity, while the same concentration showed a maximum 91% of Fe2+ chelating activity. B. paralicheniformis CamBx3 EPS (0.2 mg mL-1) demonstrated ß-glucosidase inhibition. The EPS formed a viscoelastic gel at 45°C with a maximum instantaneous viscosity of 315 Pa.s at acidic pH 5. The present study suggests that B. paralicheniformis CamBx3 could be a valuable resource for biopolymers and bioactive molecules for industrial applications. [ABSTRACT FROM AUTHOR]

In the present study, Erkence olive fruits were examined during two harvest periods and in two orchards close to the sea at low altitude (A) and far from the sea at higher altitude (B). Olives from the sea-facing northeast (NE) and land-facing southwest (SW) orientations of the trees were analyzed in terms of maturity index, total phenol content, phenolic compounds, ß-glucosidase enzyme activity, total protein content and specific enzyme activity. Climate data were collected from the trees and from the meteorology station. The maturity index, ß-glucosidase enzyme activity, total protein content and specific enzyme activity of the olives increased during the ripening period. On the other hand, the total phenol content, oleuropein and other phenolic compound contents in the olives decreased. The altitude and location of the fruits on the tree affected the ripening process. Lower altitude and sea-facing side of the tree resulted in an increased enzyme activity, thus aiding in the ripening process. [ABSTRACT FROM AUTHOR]

Bark residues from Douglas fir are an abundant resource that is currently used primarily in low-value energy recovery or is landfilled. Bark extractives are rich in diverse compounds like terpenes, fatty acids, phenols, and sugars with potential uses in a variety of high value applications. The study explores the potential of enzymatic hydrolysis to improve phenolic compounds from Douglas fir bark. It also assesses differences in chemical composition among rhytidome, phloem, and comingled bark fractions from an industrial waste pile. Phloem fractions exhibit higher yields of extractives, rhytidome fractions have elevated lignin levels, while the comingled fraction lies between the two except in ash content which was higher than in the separated fractions. Fungal decay tests with Gloeophyllum trabeum and Coniophora puteana on extract treated wood suggest potential for growth inhibition in extracts, about 58–31 % and 30–7% mass loss (in average) respectively, but due to high mass loss at low concentrations an enzymatic modification approach seems crucial for enhanced inhibition. Growth responses in whole-cell fermentation approach display variability depending on the participating microorganisms. Enzymatic hydrolysis with beta-glucosidase improved the antioxidant properties of bark extracts and holds promise for altering the chemical composition and enhancing bioactivity. [ABSTRACT FROM AUTHOR]

To reduce plant-parasitic nematodes (PPNs), an alternative solution based on amendments from selected native shrub was used to favor mainly the increase of free-living nematodes (FLNs) that include high trophic (bacterivorous, fungivorous, omnivorous, and carnivorous) nematodes and influence soil microbial activities. The consequences of the subsequent reduce of PPNs of the soil were also determined. Experiments were performed in a shade house using six treatments consisting of shrub residues from Piliostigma reticulatum (Pil), Boscia senegalensis (Bos), and Crotalaria juncea (Cro); a Straw-based Compost (Cs); a Farmer practice (FP) using Nitrogen-Phosphorus-Potassium (NPK) fertilizer; and a control (C0, no amendments). Abundances of nematodes, trophic groups, and nematodes ecological indices were determined. Enzymatic activities (ß-glucosidase and acid phosphatase) and the fluorescein diacetate hydrolysis (FDA) were also determined. Soil amended by Bos and Cro increased the density of nematodes compared to control soil or amended with NPK fertilizer. Indices of taxonomic diversity (H') did not differ significantly between the treatments. However, soils amended with Bos and Cro had the highest enrichment index (EI), and the lowest Nematode Channel Ratio (NCR), indicating that these two shrub residues exhibited a higher food web in response to the available resources provided by the fungal decomposition pathway. Moreover, Bos favored omnivorous and increased enzymatic activities. Bos induce 'bottom-up' while Cro induce 'top-down' controls on nematode populations. [ABSTRACT FROM AUTHOR]

β‐Glucosidase, a pivotal enzyme in the synthesis of various glycosides with wide applications in industries such as food, cosmetics, beverages, bioenergy, nutraceuticals, pharmaceuticals, and detergents, undergoes immobilization to enhance its utility in industrial processes and improve enzymatic properties. In this study, the conventional hybrid nanoflower synthesis method was used for β‐glucosidase‐inorganic hybrid nanoflowers. Following a comprehensive pH and temperature scanning of the nanoflowers, the influence of metallic ions and various organic solvents on hybrid nanoflowers to enhance the activity and stability of the biocatalyst were investigated. The optimal morphology for β‐Glu/NFs was determined to be 0.01 mg/mL. To obtain the optimum activity conditions, we explored pH values ranging from 3 to 8 and temperatures between 30 and 80 °C. Both the free form and β‐Glu/NFs exhibited higher activity in acidic conditions, with a significant decrease in activity observed at pH 7 and 8. β‐Glu/NFs demonstrated superior activity compared to the free enzyme in the presence of metal ions, exhibiting a remarkable 3.6‐fold increase in activity in chloroform, and approximately 2.8 and 2.7 times increase in benzene and glycerol, respectively. Moreover, it displayed over a 2‐fold increase in activity in methanol, ethanol, isopropanol, n‐butanol, acetone, and DMSO. [ABSTRACT FROM AUTHOR]

There is a growing interest in developing agricultural practices that can improve crop performance while preserving natural resources. Plant biostimulants are thought to play a role in reaching this goal, in particular by increasing the nitrogen use efficiency. However, a notable research gap exists concerning the effects of foliar applications of natural zeolites as plant biostimulants on crop performance. To address this knowledge gap, a greenhouse experiment was set up in order to study the response of maize and wheat traits, specifically the biomass productivity and nitrogen uptake, as well as the response of soil extracellular enzymes to the foliar applications of a natural zeolite in combination with different levels of nitrogen fertilization, that is, 100%, 75%, and 50% of the optimal dose. Foliar application of zeolite in wheat and maize plants led to an increase in root nitrogen concentration of about 10%, particularly at the lowest nitrogen fertilization rate. This response was accompanied by an increase in aboveground to belowground uptake nitrogen ratio. Furthermore, there was a significant reduction of about 20% in root biomass in both crops with zeolite application across the entire nitrogen fertilization gradient. These plant-level responses were associated with a significant increase in the activity of carbon-degrading and nitrogen-degrading enzymes at the soil level in response to zeolite applications. Our findings provide a compelling proof-of-concept for the beneficial effects of foliar-applied zeolite as a biostimulant for crops, emphasizing the critical need for additional field research to validate our greenhouse results. [ABSTRACT FROM AUTHOR]

Abstract The food enzyme β‐glucosidase (β‐D‐glucoside glucohydrolase; EC 3.2.1.21) is produced with the non‐genetically modified Penicillium guanacastense strain AE‐GLY by Amano Enzyme Inc. The food enzyme is intended to be used in four food manufacturing processes. Dietary exposure to the food enzyme‐total organic solids (TOS) was estimated to be up to 4.054 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90‐day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 943 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 233. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that the risk of allergic reactions by dietary exposure cannot be excluded, but the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Conifers are long-lived and slow-evolving, thus requiring effective defences against their fast-evolving insect natural enemies. The copy number variation (CNV) of two key acetophenone biosynthesis genes Ugt5/Ugt5b and βglu-1 may provide a plausible mechanism underlying the constitutively variable defence in white spruce (Picea glauca) against its primary defoliator, spruce budworm. This study develops a long-insert sequence capture probe set (Picea_hung_p1.0) for quantifying copy number of βglu-1-like, Ugt5-like genes and single-copy genes on 38 Norway spruce (Picea abies) and 40 P. glauca individuals from eight and nine provenances across Europe and North America respectively. We developed local assemblies (Piabi_c1.0 and Pigla_c.1.0), full-length transcriptomes (PIAB_v1 and PIGL_v1), and gene models to characterise the diversity of βglu-1 and Ugt5 genes. We observed very large copy numbers of βglu-1, with up to 381 copies in a single P. glauca individual. We observed among-provenance CNV of βglu-1 in P. glauca but not P. abies. Ugt5b was predominantly single-copy in both species. This study generates critical hypotheses for testing the emergence and mechanism of extreme CNV, the dosage effect on phenotype, and the varying copy number of genes with the same pathway. We demonstrate new approaches to overcome experimental challenges in genomic research in conifer defences. [ABSTRACT FROM AUTHOR]

For an agricultural region such as the Vojvodina Province in northern Serbia, soil quality monitoring is very important. Enzymatic activities are proposed as good indicators as they respond to even the slightest changes in the soil. This study aimed to analyze the enzymatic activity levels across three different soil types in Vojvodina and to examine their connection to soil chemical properties and land use. All soil types (chernozem, vertisol, solonchak) were sampled at nine locations, each with 3 field plots. The activities of acid and alkaline phosphatase, ß-glucosidase, dehydrogenase, and catalase were measured in samples, as well as the selected chemical properties. Results showed differences in enzymatic activity across different soil types and land use. The most active enzymes in vertisol were acid phosphatase and ß-glucosidase; in solonchak, it was alkaline phosphatase; in chernozem, it was dehydrogenase. A high correlation between enzymatic activities and certain soil chemical properties (pH reaction, organic matter, organic carbon, total nitrogen) was also observed, underlining the existence of a relationship between different soil components. The highest determined correlation was between acid phosphatase and pH (r=-0.7), alkaline phosphatase and total nitrogen (r=0.7), and organic matter (r=0.72); the obtained correlations were found to be statistically significant. [ABSTRACT FROM AUTHOR]

Downy mildew caused by Sclerospora graminicola is a systemic infectious disease affecting foxtail millet production in Africa and Asia. S. graminicola-infected leaves could be decomposed to a state where only the veins remain, resulting in a filamentous leaf tissue symptom. The aim of the present study was to investigate how S. graminicola influences the formation of the filamentous leaf tissue symptoms in hosts at the morphological and molecular levels. We discovered that vegetative hyphae expanded rapidly, with high biomass accumulated at the early stages of S. graminicola infection. In addition, S. graminicola could affect spikelet morphological development at the panicle branch differentiation stage to the pistil and stamen differentiation stage by interfering with hormones and nutrient metabolism in the host, resulting in hedgehog-like panicle symptoms. S. graminicola could acquire high amounts of nutrients from host tissues through secretion of ß-glucosidase, endoglucanase, and pectic enzyme, and destroyed host mesophyll cells by mechanical pressure caused by rapid expansion of hyphae. At the later stages, .S', graminicola could rapidly complete sexual reproduction through tryptophan, fatty acid, starch, and sucrose metabolism and subsequently produce numerous oospores. Oospore proliferation and development further damage host leaves via mechanical pressure, resulting in a large number of degraded and extinct mesophyll cells and, subsequently, malformed leaves with only veins left, that is, "filamentous leaf tissue." Our study revealed the S. graminicola expansion characteristics from its asexual to sexual development stages, and the potential mechanisms via which the destructive effects of S. graminicola on hosts occur at different growth stages. [ABSTRACT FROM AUTHOR]

This research investigates the potential of bioactive compounds derived from cyanobacteria as inhibitors of alphaamylase and beta-glucosidase enzymes, which are involved in starch digestion and glucose release. The study reveals strong molecular interactions between these compounds and the enzyme active sites through docking analysis. Notably, compounds such as Abietic, Anilide, Nostocarboline, Noscomin, Tanikolide, Tubercidin, Cryptophycin, and Cyanobacteria exhibit the lowest binding energies when interacting with alpha-amylase. Among them, Noscomin demonstrates the lowest docking score and binding energy against alpha-amylase, outperforming the reference compound metformin. Similarly, these compounds also display low binding energies when interacting with beta-glucosidase. The bioactive compounds from cyanobacteria show significant potential as inhibitors of alpha-amylase and beta-glucosidase, suggesting their efficacy in managing diabetes by slowing down starch digestion and controlling glucose release. Their superior binding affinities and lower binding energies, particularly Noscomin, indicate their potential in regulating blood sugar levels by interacting effectively with these enzymes. Thus, these compounds hold promise as valuable leads for developing alpha-amylase and beta-glucosidase inhibitors, contributing to the management of diabetes. Further research is required to understand the underlying mechanisms of action and assess the bioavailability, toxicity, and pharmacological potential of these cyanobacterial compounds. This investigation provides valuable insights into the potential of cyanobacterial bioactive compounds as effective candidates for the development of novel therapeutics targeting alphaamylase and beta-glucosidase enzymes in diabetes management. [ABSTRACT FROM AUTHOR]

Although the nutritional demands of animals are known to differ between immature and mature stages and between neonates from different clutches in various organisms, little evidence has been accumulated to examine this possibility in crustacean zooplankters. To this end, we examined the five digestive enzyme activities related to assimilations of carbohydrates, lipids, amino acids and phosphorus in the neonates, immature and mature individuals of panarctic Daphnia pulex. We found that all the enzyme activities were higher at lower food levels, indicating that the activities increased with decreasing supplies of the nutritional substances relative to the demands. As expected, these enzyme activities changed ontogenetically. Beta‐glucosidase activities increased with age, especially at a high food level, while the opposite pattern was found for lipase activities. A decreasing trend with age was also found in the activities of arginine and alanine amino‐peptidase and alkaline phosphatase at a low food level. These results suggest that juvenile individuals require more lipids, amino acids and phosphorus relative to the requirements of adult individuals, while the opposite pattern was true for carbohydrates. We also measured the enzyme activities of the neonates born from the 1st and 2nd clutches of the maternal individuals. The results showed that most digestive enzyme activities differed significantly between the neonates from the first and second clutches depending on the food levels. The present study confirmed that the nutritional demands differ not only between mature and immature individuals in Daphnia pulex and suggests that the mature individuals invest nutrients and energy reserves differently to neonates of the different clutches depending on their nutritional conditions. [ABSTRACT FROM AUTHOR]

The defensive-offensive associations between algae and herbivores determine marine ecology. Brown algae utilize phlorotannin as their chemical defense against the predator Aplysia kurodai, which uses ß-glucosidase (akuBGL) to digest the laminarin in algae into glucose. Moreover, A. kurodai employs Eisenia hydrolysis-enhancing protein (EHEP) as an offense to protect akuBGL activity from phlorotannin inhibition by precipitating phlorotannin. To underpin the molecular mechanism of this digestive-defensive-offensive system, we determined the structures of the apo and tannic acid (TNA, a phlorotannin analog) bound forms of EHEP, as well as the apo akuBGL. EHEP consisted of three peritrophin-A domains arranged in a triangular shape and bound TNA in the center without significant conformational changes. Structural comparison between EHEP and EHEP-TNA led us to find that EHEP can be resolubilized from phlorotannin precipitation at an alkaline pH, which reflects a requirement in the digestive tract. akuBGL contained two GH1 domains, only one of which conserved the active site. Combining docking analysis, we propose the mechanisms by which phlorotannin inhibits akuBGL by occupying the substrate-binding pocket, and EHEP protects akuBGL against this inhibition by binding with phlorotannin to free the akuBGL pocket. [ABSTRACT FROM AUTHOR]

Trichoderma erinaceum is a filamentous fungus that was isolated from decaying sugarcane straw at a Brazilian ethanol biorefinery. This fungus shows potential as a source of plant cell wall-degrading enzymes (PCWDEs). In this study, we conducted a comprehensive multiomics investigation of T. erinaceum to gain insights into its enzymatic capabilities and genetic makeup. Firstly, we performed genome sequencing and assembly, which resulted in the identification of 10,942 genes in the T. erinaceum genome. We then conducted transcriptomics and secretome analyses to map the gene expression patterns and identify the enzymes produced by T. erinaceum in the presence of different substrates such as glucose, microcrystalline cellulose, pretreated sugarcane straw, and pretreated energy cane bagasse. Our analyses revealed that T. erinaceum highly expresses genes directly related to lignocellulose degradation when grown on pretreated energy cane and sugarcane substrates. Furthermore, our secretome analysis identified 35 carbohydrate-active enzymes, primarily PCWDEs. To further explore the enzymatic capabilities of T. erinaceum, we selected a β-glucosidase from the secretome data for recombinant production in a fungal strain. The recombinant enzyme demonstrated superior performance in degrading cellobiose and laminaribiose compared to a well-known enzyme derived from Trichoderma reesei. Overall, this comprehensive study provides valuable insights into both the genetic patterns of T. erinaceum and its potential for lignocellulose degradation and enzyme production. The obtained genomic data can serve as an important resource for future genetic engineering efforts aimed at optimizing enzyme production from this fungus.

The Lactococcus taiwanensis strain K_LL004 was isolated from the gut of a grasshopper (Oxya chinensis sinuosa) collected from local farm in Korea. L. taiwanensis strain K_LL004 is the functional probiotic candidate with an ability to hydrolyse plant polysaccharides. The complete genome of the L. taiwanensis strain K_LL004 contains one circular chromosome (1,995,099 bp) with a guanine + cytosine (GC) content of 38.8%. Moreover, 1,929 Protein-coding sequence, 19 rRNA genes, and 62 tRNA genes were identified based on results of annotation. L. taiwanensis strain K_LL004 has a gene, which encodes hydrolytic enzymes such as beta-glucosidase and beta-xylosidase, that hydrolyzes plant polysaccharides.

The objective of this study, is characterization of partially purified beta-glucosidase (BGS) extracted from apricot defatted seeds powder. The enzyme was extracted using sodium phosphate solution (0.2M, pH 7) containing 2% sodium chloride with mixing ratio of (1:5 solid: liquid) for extraction period of 3 hours at 4C. The crude extract was precipitated by ammonium sulfate at 20-70% saturation, dialyzed then concentrated. The specific activity of the enzyme, folds of purification and the enzymatic yield were 5.45 units/mg, 7.8 fold and 84.5%, respectively. The results showed that the optimum conditions for the enzyme activity were at pH 5.5 and 6.5, the optimum temperature for the enzyme’s activity was 40°C and 55°C, while the thermal stability of the enzyme was between 25-45°C when the enzyme retained more than 90% of its activity over 30 minutes incubation at the optimum pH for activity, and retained 88% of its activity at 50° C. The results also revealed the inhibitory and activator effects of minerals and chemical compounds on the BGS activity. [ABSTRACT FROM AUTHOR]

Beans (Vicia faba) contain some anti-nutritional compounds, especially vicine and convicine and their non-sugar derivatives, divicine and isouramil, which are responsible for the occurrence of Favism in people with hereditary deficiency of Glucose-6-phosphate dehydrogenase (G6PD). These study objective was estimation of vicine and convicine content and their distribution in dry and green bean parts during the growth stages, and the effect of beta-glucosidase from defatted apricot seed powder and some home-based processing conditions on elimination of these compounds from the bean. The results showed that the content of vicine & convicine in bean ranged from 22.7 mg/100 g in the seedling stage to 46.8 mg/100 g in the pods stage at 139 days. As for vicine, it ranged from 12.6 mg /100g to 29 mg/100g at the above stages respectively. While for convicine it ranged from 10.1 mg/100g to 17.8 mg/100 respectively. The enzymatic treatments showed significant elimination of those compounds as compared to non-enzymatic treatments. Additionally, other factors such as soaking temperature, pH, and time as well as cooking process had a positive effect on the elimination of those glucosides. [ABSTRACT FROM AUTHOR]

It is well established that land use change (LUC) can impact soil organic carbon (SOC) in tropical regions, but the long-term effects of LUC on soil quality and C cycling remain unclear. Here, we evaluated how LUC affects soil C cycling in the Amazon region using a 100-year observational chronosequence spanning primary forest-to-pasture conversion and subsequent secondary forest succession. We found a surprising increase in topsoil SOC concentrations 60 years following conversion, despite major losses (> 85%) of forest-derived SOC within the first 25 years. Shifts in molecular composition of SOC, identified with diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy, occurred in tandem with a significant decline in permanganate-oxidizable C (POXC) and β-glucosidase activity (per unit SOC), interpreted as a deceleration of soil C cycling after pasture grasses became the dominant source of C inputs to soil. Secondary forest succession caused rapid reversal to conditions observed under primary forest for β-glucosidase activity but not for SOC molecular composition (DRIFT spectroscopy), reflecting a long-lasting effect of LUC on soil C cycling. Our results show that rapid changes in the origin of SOC occur following deforestation with legacy effects on some indicators of C cycling (e.g. enzyme activity) but not others (e.g. molecular composition). This approach offers mechanistic understanding of LUC in the Amazon basin and can be used to help explain conflicting reports on how deforestation impacts SOC in the region.

Lysosomes are the catabolic center of the cell. Limitations of many lysosomal tracers include low specificity and lack of reliable physiological readouts for changes in growth factor-regulated lysosomal activity. The imaging-based protocols described here provide insights at the cellular level to quantify functions essential to lysosomal biology, including β-glucosidase enzymatic cleavage, active Cathepsin D, and pH regulation in real time. These optimized protocols, applied in different cell types and pathophysiologic contexts, provide useful tools to study lysosome function in cultured living cells. For complete details on the use and execution of this protocol, please refer to Albrecht et al. (2020).

Background and aims: Colletotrichum graminicola (Glomerellaceae, Glomerellales), the causal agent of maize (Zea mays) anthracnose, as many other fungal pathogens, relies on its battery of cell wall degrading enzymes (CWDEs) to make its way through the cell walls of the host, and thus the production of these enzymes is considered a virulence factor. The aim of this work was to investigate if there is intraspecific variability in growth and in vitro production of several extracellular CWDEs among Argentinean fungal isolates of C. graminicola. M&M: Eight isolates of C. graminicola were tested in vitro to evaluate growth capacity and polygalacturonase, polymethylgalacturonase, ß-glucosidase and laccase production, using two different liquid culture media. Results: Polygalacturonase, polymethylgalacturonase and ß-glucosidase production greatly varied among isolates and culture media. Laccase activity was detected only in three isolates. Utmost enzymatic titres attained were respectively 250, 280, 45 and 63 U/l. The observed intraspecific variability in CWDEs in vitro production is consistent with the high variability found at genetic level when assessing ISSR markers. Conclusions: The isolates of C. graminicola evaluated showed notable differences in CWDEs production, not associated with a differential growth. This indicates a large intraspecific variability, which might be considered when choosing a method to deal with this pathogen. [ABSTRACT FROM AUTHOR]

A novel endophytic fungus producing beta-glucosidase was isolated and characterized from pigeon pea (Cajanus cajan [L.] Millsp.), which has excellent properties in converting ginsenoside Rb1 to ginsenoside Rd in Panax notoginseng. According to the 16S rDNA gene sequence, the G11-7 strain was identified as Fusarium proliferatum, and the accession number KY303906 was confirmed in GenBank. The G11-7 immobilized spores, in which the activity of beta-glucosidase could reach 0.95 U/mL, were co-cultured with P. notoginseng plant material to obtain a continuous beta-glucosidase supply for the biotransformation of ginsenoside Rb1 to Rd. Under the liquid–solid ratio (20:1), initial pH (6.0), and temperature (30 °C) constituents, the maximum ginsenoside Rd yield was obtained as 9.15 ± 0.65 mg/g, which was 3.67-fold higher than that without fungal spore co-culture (2.49 ± 0.98 mg/g). Furthermore, immobilized G11-7 spores showed significant beta-glucosidase producing ability which could be recovered and reused for 6 cycles. Overall, these results suggested that immobilized G11-7 offered a promising and effective approach to enhance the production of ginsenoside Rd for possible nutraceutical and pharmaceutical uses. [ABSTRACT FROM AUTHOR]

Programmed cell death (PCD) in lateral root caps (LRCs) is crucial for maintaining root cap functionality. Endoplasmic reticulum (ER) bodies play important roles in plant immunity and PCD. However, the distribution of ER bodies and their communication with vacuoles in the LRC remain elusive. In this study, we investigated the ultrastructure of LRC cells of wild-type and transgenic Arabidopsis lines using an auto-acquisition transmission electron microscope (TEM) system and high-pressure freezing. Gigapixel-scale high-resolution TEM imaging of the transverse and longitudinal sections of roots followed by three-dimensional imaging identified sausage-shaped structures budding from the ER. These were subsequently identified as ER bodies using GFPh transgenic lines expressing green fluorescent protein (GFP) fused with an ER retention signal (HDEL). Immunogold labeling using an anti-GFP antibody detected GFP signals in the ER bodies and vacuoles. The fusion of ER bodies with vacuoles in LRC cells was identified using correlative light and electron microscopy. Imaging of the root tips of a GFPh transgenic line with a PYK10 promoter revealed the localization of PYK10, a member of the β-glucosidase family with an ER retention signal, in the ER bodies in the inner layer along with a fusion of ER bodies with vacuoles in the middle layer and collapse of vacuoles in the outer layer of the LRC. These findings suggest that ER bodies in LRC directly transport β-glucosidases to the vacuoles, and that a subsequent vacuolar collapse triggered by an unknown mechanism releases protective substances to the growing root tip to protect it from the invaders. [ABSTRACT FROM AUTHOR]

Within the Svalbard archipelago, Kongsfjorden is an important marine ecosystem that is recognised as one of the main representative Arctic glacial fjords. Prokaryotic organisms are key drivers of important ecological processes such as carbon fluxes, nutrient mineralisation, and energy transfer, as well as sentinels of environmental pollution, especially in sediments, that are a repository of contaminants. In some areas of the Arctic, the structure and metabolic activity of the microbial community in the organic matter turnover and globally in the functioning of the benthic domain are mostly still unknown. A snapshot of the main microbial parameters such as bacterial abundance (by microscopic and plate counts), structure (by 16S rRNA sequencing), and metabolic activity was provided in Ny-Ålesund harbour, contextually in seawater and sediment samples. Fluorogenic substrates were used to assess the microbial ability to utilise organic substrates such as proteins, polysaccharides, and organic phosphates through specific enzymatic assays (leucine aminopeptidase—LAP, beta-glucosidase—ß-GLU, and alkaline phosphatase—AP, respectively). The metabolic profiles of psychrophilic heterotrophic bacterial isolates were also screened using a qualitative assay. The phylogenetic analysis of the microbial community revealed that Proteobacteria prevailed among the observed taxonomic groups. Several of the observed sequences were assigned to clones found in harbours, microbial biofilms, antifouling paints, or oil-polluted facilities of cold environments, highlighting a signature of human pressure on the polar habitat of Ny-Ålesund harbour. [ABSTRACT FROM AUTHOR]

Filamentous fungi are intensively used for producing industrial enzymes, including lignocellulases. Employing insoluble cellulose to induce the production of lignocellulases causes some drawbacks, e.g., a complex fermentation operation, which can be overcome by using soluble inducers such as cellobiose. Here, a triple β-glucosidase mutant of Neurospora crassa, which prevents rapid turnover of cellobiose and thus allows the disaccharide to induce lignocellulases, was applied to profile the proteome responses to cellobiose and cellulose (Avicel). Our results revealed a shared proteomic response to cellobiose and Avicel, whose elements included lignocellulases and cellulolytic product transporters. While the cellulolytic proteins showed a correlated increase in protein and mRNA levels, only a moderate correlation was observed on a proteomic scale between protein and mRNA levels (R2 = 0.31). Ribosome biogenesis and rRNA processing were significantly overrepresented in the protein set with increased protein but unchanged mRNA abundances in response to Avicel. Ribosome biogenesis, as well as protein processing and protein export, was also enriched in the protein set that showed increased abundance in response to cellobiose. NCU05895, a homolog of yeast CWH43, is potentially involved in transferring a glycosylphosphatidylinositol (GPI) anchor to nascent proteins. This protein showed increased abundance but no significant change in mRNA levels. Disruption of CWH43 resulted in a significant decrease in cellulase activities and secreted protein levels in cultures grown on Avicel, suggesting a positive regulatory role for CWH43 in cellulase production. The findings should have an impact on a systems engineering approach for strain improvement for the production of lignocellulases.IMPORTANCE Lignocellulases are important industrial enzymes for sustainable production of biofuels and bio-products. Insoluble cellulose has been commonly used to induce the production of lignocellulases in filamentous fungi, which causes a difficult fermentation operation and enzyme loss due to adsorption to cellulose. The disadvantages can be overcome by using soluble inducers, such as the disaccharide cellobiose. Quantitative proteome profiling of the model filamentous fungus Neurospora crassa revealed cellobiose-dependent pathways for cellulase production, including protein processing and export. A protein (CWH43) potentially involved in protein processing was found to be a positive regulator of lignocellulase production. The cellobiose-dependent mechanisms provide new opportunities to improve the production of lignocellulases in filamentous fungi.

Amygdalin is known as a chemical compound derived from various fruits. The glycosides existing in this plant have been historically utilized as an anticancer agent. This review presented an overview of amygdalin and its onco-immunity and other therapeutic medical applications. A literature search for studies relating to amygdalin and cancer treatment was carried out using PubMed and Google Scholar. Combinations of the following terms were used in the search strategies: "amygdalin," "rhodanese," "cyanide," "cyanogenic," "hypothiocyanite," "mandelonitrile," "glucosides," "cancer," "apoptosis," and "cytotoxicity," combined with a cancer term such as "seed," "almond," or "apricot," "cancer + cell line, antiproliferation or inhibition," "BAX From the March 3, 1981 until the April 15, 2021, all of the English-language papers were evaluated based on the inclusion criteria. Publications included reviews, chapters from books, and original research papers. The FDA prohibits Amygdalin from medical usage as an anticancer treatment due to a lack of proof of cure in cancer cases. When this natural-based compound is used with conditional chemotherapeutic medicines causes synergistic effects. Besides, amygdalin is used to manage asthma, improve the immune system, induce apoptosis in human renal fibroblasts, and inhibit hyperglycemia. Various medical uses of amygdalin have been found such as managing asthma, improving the immune system, inducing apoptosis in human renal fibroblasts, and inhibiting hyperglycemia. More effective in vitro and review studies are required to elucidate the exact role of this herb in medical applications. [Display omitted] [ABSTRACT FROM AUTHOR]

Microwave irradiation at different frequencies gave molecular selective effects, namely higher frequency microwave effects for waters while lower frequency effects for ions. We already reported that 2.45 GHz and 5.80 GHz microwave irradiation gave different results for a hydrolysis reaction by thermostable ß-Glucosidase HT1. Here, we designed and made a reactor, employed 400 MHz microwave irradiation, and studied the effectiveness of 400 MHz microwave for HT1 reaction, then 400 MHz and 2.45 GHz had the ability to accelerate HT1 reaction. In consideration of the general mechanism of enzymatic glycoside hydrolysis, our results would be reasonable if ions are key reaction species because 400 MHz microwave activated ions selectively. In addition, the phenomenon that 400 MHz microwave would not affect water molecules by dielectric heating might contribute the enzyme stability. This report should support that microwave is not only a tool to heat reactions efficiently but also can bring unique effects for reactions. [ABSTRACT FROM AUTHOR]

The production of second-generation bioethanol has several challenges, among them finding cheap and efficient enzymes for a sustainable process. In this work, we analyzed two native fungi, Cladosporium cladosporioides and Penicillium funiculosum, as a source of cellulolytic enzyme production, and corn stover, wheat bran, chickpeas, and bean straw as a carbon source in two fermentation systems: submerged and solid fermentation. Corn stover was selected for cellulase production in both fermentation systems, because we found the highest enzymatic activities when carboxymethyl cellulase activity (CMCase) was assessed using CMC as substrate. C. cladosporioides showed the highest CMCase activity (1.6 U/mL), while P. funiculosum had the highest filter paper activity (Fpase) (0.39 U/mL). The ß-glucosidase activities produced by both fungi were similar in submerged fermentation using corn stover as substrate. Through in-gel zymography, three polypeptides with cellulolytic activities were identified in each fungus: with molecular weights of ~ 38, 45 and 70 kDa in C. cladosporioides and ~ 21, 63 and 100 kDa in P. funiculosum. The best results for saccharification (10.11 g/L of reducing sugars) of diluted acid pretreated corn stover were obtained after 36 h of the hydrolytic process at pH 5 and 50 °C using the enzyme extract of P. funiculosum. This is the first report of cellulase identification in C. cladosporioides and the saccharification of corn stover using enzymes of this fungus. Enzymatic extracts of C. cladosporioides and P. funiculosum obtained from low-cost lignocellulosic biomass have great potential for use in the production of second-generation bioethanol. [ABSTRACT FROM AUTHOR]

The present study aimed to evaluate the effect of pure and mixed culture fermentations with autochtonous yeast strains Torulaspora delbrueckii and Saccharomyces cerevisiae on physico-chemical and sensory qualities of Fetească albă and Fetească neagră young wines, at pilot scale. Yeast strains were isolated during different stages of spontaneous alcoholic fermentation and selected for their potential role in winemaking: 26 strains, 17 Saccharomyces cerevisiae strains and 9 non-Saccharomyces strains, from genera Candida, Torulaspora and Debaryomyces, were evaluated in terms of extracellular enzymatic activity, fermentative and technological characteristics. From these, two Saccharomyces cerevisiae strains were selected for esterase activity, and two non-Saccharomyces strains, Torulaspora delbrueckii species were selected for ß-glucosidase and esterase activity. Different yeast strains influenced the physicochemical characteristics of the wines. The sensory qualities of the young wines produced with mixed cultures were positively influenced. [ABSTRACT FROM AUTHOR]

Divicine is an active pyrimidine aglycone, generated from vicine by the enzyme ß-glucosidase upon ingestion of fava beans. In this study, we investigated the effect of divicine on cultured human umbilical vein endothelial cells (HUVECs) and explored the potential mechanisms. Incubation HUVECs with 18.5-85.1 µM divicine resulted in a concentration and time dependent decrease of cell viability, followed by decrease of cellular reduced glutathione, as well as increase of reactive oxygen species (ROS), malondialdehyde (MDA), and labile iron pool. Transmission electron microscopy confirmed that the divicine treated HUVECs' mitochondria had shrunk. Importantly, the administration of desferrioxamine, an iron chelator, to the divicine treated HUVECs significantly reduced iron overload and cell death and decreased cellular ROS and MDA. These results demonstrated that divicine could cause damaging of endothelial cells, and ferroptosis might be involved in divicine induced HUVECs injury, reminding long term ingestion of fava beans might be harmful to vascular system, especially for those suffering from glucose 6-phosphate dehydrogenase deficiency. [ABSTRACT FROM AUTHOR]

Genome resequencing is an efficient strategy for associating mutant phenotypes with physical genomic loci (Baker 2009). A pilot study of this approach demonstrated that the Neurospora crassa genetic map was critical in narrowing the possible candidate mutations in a strain to a small number in a limited, defined region of the genome (McCluskey et al.2011). In this study, we utilize a resequencing strategy to identify the mutations underlying the gluc-1 and gluc-2 genes in N. crassa. [ABSTRACT FROM AUTHOR]

A regression-based approach was used to test the suitability of a range of parameters, including total prokaryotic cell abundance and biomass, as lipopolysaccharide (LPS) content, and exoenzymatic activities (leucine aminopeptidase, LAP, beta-glucosidase, ß-G, and alkaline phosphatase, AP) as predictors of heterotrophic prokaryotic production (HPP) in the Ross Sea epipelagic waters. A close association between HPP and protein hydrolysis mediated by enzymatic activity (LAP), and to a lower significance level with the other variables, was recorded. Three multiple regression equations were developed from two microbial datasets collected during middle austral summer periods. All showed a good predictive ability for HPP, and this was further validated through a comparison between the predicted and the observed HPP values. The obtained regression equations proved to represent a promising example of empirical models for further predictive studies in the Ross Sea where—through the incorporation of additional microbiological and environmental parameters—the developed models could find a practical application to cover the entire austral summer period. [ABSTRACT FROM AUTHOR]

The insufficient of lignocellulose degradation enzymes, such as cellulase and hemicellulase, is the major obstacle that hinders the bioconversion of lignocellulosic biomass to monosaccharides, especially during the woody biomass hydrolysis process. The addition of additives has received significant attention due to their enhancement of the enzymatic degradation efficiency of lignocellulose. In the present study, a combination of organic alcohols and a biosurfactant could synergistically enhance the saccharification of the cellulose substrate of Avicel, as well as that of pretreated poplar. Results showed that compound additives can greatly improve the conversion rate of enzymatic hydrolysis. The combination of 0.1% (v/v) n-decanol and 1% (v/v) sophorolipid dramatically increased the poplar enzymatic conversion rate from 17.9% to 85%, improving it by 67.1%. Enzyme-rich Hypocrea sp. W63 was fermented to obtain beta-glucosidase (BGL) and xylanase (XYL), which were used as auxiliary enzymes during enzymatic hydrolysis. It was found that the effects of such a combination of additives improved the filter paper activity, stability, and longevity, helping in the recovery of the cellulase cocktail. The compound additives associated with the commercial cellulase and Hypocrea sp. W63 enzyme solution formed an excellent formula for improving the stability of BGL and XYL. The results provide insight into compound additives and the use of a cellulase and auxiliary enzyme cocktail to improve enzymatic hydrolysis for lignocellulose conversion into biofuels. [ABSTRACT FROM AUTHOR]

Trichodermareesei is a widely used host for producing cellulase and hemicellulase cocktails for lignocellulosic biomass degradation. Here, we report a genetic modification strategy for industrial T.reesei that enables enzyme production using simple glucose without inducers, such as cellulose, lactose and sophorose. Previously, the mutated XYR1V821F or XYR1A824V was known to induce xylanase and cellulase using only glucose as a carbon source, but its enzyme composition was biased toward xylanases, and its performance was insufficient to degrade lignocellulose efficiently. Therefore, we examined combinations of mutated XYR1V821F and constitutively expressed CRT1, BGLR, VIB1, ACE2, or ACE3, known as cellulase regulators and essential factors for cellulase expression to the T.reesei E1AB1 strain that has been highly mutagenized for improving enzyme productivity and expressing a ß-glucosidase for high enzyme performance. The results showed that expression of ACE3 to the mutated XYR1V821F expressing strain promoted cellulase expression. Furthermore, co-expression of these two transcription factors also resulted in increased productivity, with enzyme productivity 1.5-fold higher than with the conventional single expression of mutated XYR1V821F. Additionally, that productivity was 5.5-fold higher compared to productivity with an enhanced single expression of ACE3. Moreover, although the DNA-binding domain of ACE3 had been considered essential for inducer-free cellulase production, we found that ACE3 with a partially truncated DNA-binding domain was more effective in cellulase production when co-expressed with a mutated XYR1V821F. This study demonstrates that co-expression of the two transcription factors, the mutated XYR1V821F or XYR1A824V and ACE3, resulted in optimized enzyme composition and increased productivity. [ABSTRACT FROM AUTHOR]

The establishment of soybean seedlings is sensitive to flooding, and colocalization of proteins and quantitative trait loci (QTLs) advances to narrow down the hub genes. To reveal soybean seedling tolerance to flooding, Qihuang34 (QH34), Jidou17(JD17), and their recombinant inbred lines were used in this study. Severe plant death, shorter root length, and more stress-induced proteins were observed in JD7 compared to QH34 under flooding. The major functional categories of proteins altered by flooding were similar in both cultivars, and formed a cohesive network. Protein abundance and gene expression of ribosomal protein S3 decreased in flooded JD17 compared to QH34. Through proteomics and QTL mapping, beta-glucosidase 44 and dehydrin were involved in flood tolerance. Beta-glucosidase 44 abundance, beta-glucosidase activity, and glucose content decreased in QH34 compared to JD17 during flooding. The dehydrin contained the cis- acting elements of methyl jasmonate (Me-JA) and gibberellin (GA), accompanied by increased levels of Me-JA and GA 3 in flooded QH34 and JD17, respectively. These findings suggest that integrating proteomics with QTL is advantageous to identify genes associated with flood tolerance, proposing that enhanced protein homeostasis and cell wall stiffening in root tips may confer soybean seedling tolerance to flooding via GA and JA signals. • Soybean seedling Qihuang34 grew better compared to Jidou17 at flood. • Proteomics and QTL mapping were utilized to reveal flooding tolerance. • Transcripts and abundance of ribosomal protein S3 only reduced in Jidou17 at flood. • Beta-glucosidase 44 and Mat1 were identified as flooding tolerant genes. • Cell wall remodeling and GA and JA signals facilitate root-tip growth at flood. [ABSTRACT FROM AUTHOR]

Indigo is an ancient dye uniquely capable of producing the signature tones in blue denim; however, the dyeing process requires chemical steps that are environmentally damaging. We describe a sustainable dyeing strategy that not only circumvents the use of toxic reagents for indigo chemical synthesis but also removes the need for a reducing agent for dye solubilization. This strategy utilizes a glucose moiety as a biochemical protecting group to stabilize the reactive indigo precursor indoxyl to form indican, preventing spontaneous oxidation to crystalline indigo during microbial fermentation. Application of a β-glucosidase removes the protecting group from indican, resulting in indigo crystal formation in the cotton fibers. We identified the gene coding for the glucosyltransferase PtUGT1 from the indigo plant Polygonum tinctorium and solved the structure of PtUGT1. Heterologous expression of PtUGT1 in Escherichia coli supported high indican conversion, and biosynthesized indican was used to dye cotton swatches and a garment.

The beta-glucosidase BglC fulfills multiple functions in both primary metabolism and induction of pathogenicity of Streptomyces scabiei, the causative agent of common scab in root and tuber crops. Indeed, this enzyme hydrolyzes cellobiose and cellotriose to feed glycolysis with glucose directly and modifies the intracellular concentration of these cello-oligosaccharides, which are the virulence elicitors. The inactivation of bglC led to unexpected phenotypes such as the constitutive overproduction of thaxtomin A, the main virulence determinant of S. scabiei. In this work, we reveal a new target substrate of BglC, the phytoalexin scopolin. Removal of the glucose moiety of scopolin generates scopoletin, a potent inhibitor of thaxtomin A production. The hydrolysis of scopolin by BglC displayed substrate inhibition kinetics, which contrasts with the typical Michaelis–Menten saturation curve previously observed for the degradation of its natural substrate cellobiose. Our work, therefore, reveals that BglC targets both cello-oligosaccharide elicitors emanating from the hosts of S. scabiei, and the scopolin phytoalexin generated by the host defense mechanisms, thereby occupying a key position to fine-tune the production of the main virulence determinant thaxtomin A.

Bermudagrass (Cynodon spp.) is one of the most widely distributed warm-season grasses globally. The growth habits and plant type of bermudagrass are strongly associated with the applied purpose of the landscape, livestock, and eco-remediation. Therefore, persistent efforts are made to investigate the genetic basis of plant type and growth habits of bermudagrass. Here, we dissect the genetic diversity of 91 wild bermudagrass resources by genome-wide association studies (GWAS) combined with weighted gene co-expression analysis (WGCNA). This work is based on the RNA-seq data and the genome of African bermudagrass (Cynodon transvaalensis Burtt Davy). Sixteen reliable single-nucleotide polymorphisms (SNPs) in transcribed regions were identified to be associated with the plant height and IAA content in diverse bermudagrass by GWAS. The integration of the results from WGCNA indicates that beta-glucosidase 31 (CdBGLU31) is a candidate gene underlying a G/A SNP signal. Furthermore, both qRT-PCR and correlation coefficient analyses indicate that CdBGLU31 might play a comprehensive role in plant height and IAA biosynthesis and signal. In addition, we observe lower plant height in Arabidopsis bglu11 mutants (homologs of CdBGLU31). It uncovers the breeding selection history of different plant types from diverse bermudagrass and provides new insights into the molecular function of CdBGLU31 both in plant types and in IAA biosynthetic pathways. [ABSTRACT FROM AUTHOR]

Anacardium occidentale L. is used throughout the world to treat type 2 diabetes. In Portugal, a traditional herbal preparation made with stem bark of this species (AoBTHP) has been used for more than 30 years to treat this pathology. The AoBTHP was standardized on total phenolic content, and its hypoglycemic activity was assessed using db/db mice (n = 26) for 92 days. Three doses (40.2, 71.5, and 127.0 mg/kg/day, per os) were tested, and glibenclamide (5 mg/kg/day) was used as positive control. During the study, glycemia was measured under non-fasting or fasting states. In sequence, thin-layer chromatography bioautographic assays were used for the detection of possible alpha- and beta-glucosidase inhibitors. A significant hypoglycemic effect in fasting glycemia in days 31 and 57 was observed with the three tested doses. The 71.5 mg/kg and 127.0 mg/kg AoBTHPs significantly reduced non-fasting glycemia on day 24. The highest dose showed the most significant hypoglycemic effect. Gallic acid was identified as the major alpha- and beta-glucosidase inhibitor. The 127 mg/kg/day AoBTHP dose showed a greater glucose-lowering effect than glibenclamide. For the first time, a standardized AoBTHP was tested using an in vivo diabetes model, and its usage was preclinically validated for type 2 diabetes treatment. The hypoglycemic activity of an AoBTHP can be related to the presence of alpha- and beta-glucosidase inhibitors, such as gallic acid, but other mechanisms can also be involved. [ABSTRACT FROM AUTHOR]

A novel β-glucosidase (Thglu3) was identified from Thermotoga sp. which had biotransformation activity for notoginsenoside R1 (NR-R1). Sequence analysis of Thglu3 revealed that it could be classified into glycoside hydrolase family 3 (GH3). The gene encoding a 719-amino acid protein was cloned and expressed in Escherichia coli. The recombinant enzyme was purified, and its molecular weight was approximately 81 kDa. The recombinant Thglu3 exhibited an optimal activity at 75 °C and pH 6.4. The β-glucosidase had high selectivity for cleaving the outer glucose moiety at the C20 position of NR-R1, which produced the more pharmacologically active notoginsenoside R2 (NR-R2). Under the optimal reaction conditions for gram-scale production, 30 g NR-R1 was transformed to NR-R2 using 20 g crude enzyme at pH 6.4 and 75 °C within 1 h with a molar yield of 93%. This study was the first report of the highly efficient and selective gram-scale transformation of NR-R2 from NR-R1 by a thermophilic β-glucosidase. [ABSTRACT FROM AUTHOR]

Understanding how fertilizer application (particularly N, the most used chemical fertilizer worldwide) interacts with soil microbes is important for the development of best management practices that target improved microbial activity to enhance sustainable food production. This study was conducted to determine whether urea N rate and time of application to maize (Zea mays) influenced soil enzyme activity. Enzyme activity was determined by monitoring fluorescein diacetate (FDA) hydrolysis, ß-glucosidase, acid-phosphomonoesterase, and arylsulfatase activities. Experiments were conducted from 2014 through 2016 to compare single (fall or spring applications) and split applications of N at varying N rates under irrigation (Becker) and rainfed conditions (Lamberton and Waseca) in MN, USA. Nitrogen rates varied by location and were based on University of Minnesota guidelines. Soil samples were collected seven times each season. Nitrogen application split into two applications increased FDA activity by 10% compared with fall and spring applied N at Waseca. Fall or spring N application decreased arylsulfatase activity by 19% at Becker and by between 13% and 16% at Lamberton. ß-Glucosidase and acid-phosphomonoesterase activities were unaffected by N application. Sampling time and year had the greatest impact on enzyme activity, but the results varied by location. A negative linear relationship occurred between FDA and ß-glucosidase activity at all three sites. In summary, urea N application had small effects on enzyme activity at the sites studied, suggesting that some form of organic N could be more important than the ammonium provided by urea. [ABSTRACT FROM AUTHOR]

Beta-glucosidase (Bgl) is an enzyme with considerable food, beverage, and biofuel processing potential. However, as many Bgls are inhibited by their reaction end product glucose, their industrial applications are greatly limited. In this study, a novel Bgl gene (Bgl1973) was cloned from Leifsonia sp. ZF2019 and heterologously expressed in E. coli. Sequence analysis and structure modeling revealed that Bgl1973 was 748 aa, giving it a molecular weight of 78 kDa, and it showed high similarity with the glycoside hydrolase 3 (GH3) family Bgls with which its active site residues were conserved. By using pNPGlc (p-nitrophenyl-β-D-glucopyranoside) as substrate, the optimum temperature and pH of Bgl1973 were shown to be 50 °C and 7.0, respectively. Bgl1973 was insensitive to most metal ions (12.5 mM), 1% urea, and even 0.1% Tween-80. This enzyme maintained 60% of its original activity in the presence of 20% NaCl, demonstrating its excellent salt tolerance. Furthermore, it still had 83% residual activity in 1 M of glucose, displaying its outstanding glucose tolerance. The Km, Vmax, and kcat of Bgl1973 were 0.22 mM, 44.44 μmol/min mg, and 57.78 s−1, respectively. Bgl1973 had a high specific activity for pNPGlc (19.10 ± 0.59 U/mg) and salicin (20.43 ± 0.92 U/mg). Furthermore, molecular docking indicated that the glucose binding location and the narrow and deep active channel geometry might contribute to the glucose tolerance of Bgl1973. Our results lay a foundation for the studying of this glucose-tolerant β-glucosidase and its applications in many industrial settings. Key points: • A novel β-glucosidase from GH3 was obtained from Leifsonia sp. ZF2019. • Bgl1973 demonstrated excellent glucose tolerance. • The glucose tolerance of Bgl1973 was explained using molecular docking analysis. [ABSTRACT FROM AUTHOR]

The study aimed to assess the effects of fungicide QuadrisR on activities of soil enzymes contributed to soil nutrient turnover. A batch laboratory experiment with QuadrisR-amended (0 mg/kg ds (dry soil) - 35.00 mg/kg ds) loamy sand soil was conducted, and shifts in soil physical environments and enzyme activities (beta-glucosidase, urease, acid and alkaline phosphatases, arylsulfatase and dehydrogenase) were evaluated on experimental days 1, 30, 60, 90 and 120. The results indicated that QuadrisR changed both soil properties and enzyme activities. The most sensitive environmental parameter to fungicide input was soil pH. The most suscaptable to QuadrisR enzymes were dehydrogenase and arylsulfatase, and the most resistant - urease. The mean overall dehydrogenase activity decreased by 33%, whereas the profile of arylsulfatase activity tended to permanent decrease over time. The general pattern of enzyme responses to QuadrisR was an immediate-early (days 1 - 30) decline of enzyme activities after fungicide application, except that of arylsulfatase. Beta-glucosidase manifested a temporal profile of steady-state stimulation under the lowest (2.90 mg/kg ds) and low sensitivity to the higher (14.65 mg/kg ds and 35.00 mg/kg ds) fungicide concentrations. [ABSTRACT FROM AUTHOR]