Search Results (505)

This study optimized supercritical fluid extraction (SC-CO₂) parameters for Boswellia sacra leaves to evaluate how extraction thermodynamics influence subsequent pharmacological bioactivity. Using a multi-parametric approach, extracts obtained at 40 °C and 80 °C (100 bar) were profiled via HPLC-DAD and evaluated using in vitro antimicrobial, pancreatic lipase inhibition, and Caco-2 cell antiproliferative assays. HPLC analysis revealed that the 40 °C threshold preserved chemical diversity (17 active peaks, retaining higher caffeic acid and rutin concentrations), whereas 80 °C conditions caused severe compound degradation. Consequently, the 40 °C extracts exhibited significantly enhanced biomedical potencies over the 80 °C variants, demonstrating superior microbiostatic activity against Gram-positive bacteria and Candida albicans (MIC = 15.62 µg/mL), stronger porcine pancreatic lipase inhibition (IC₅₀ = 17.21 µg/mL), and greater antiproliferative efficacy against human colorectal adenocarcinoma cells (IC₅₀ = 109.47 µg/mL). Ultimately, maintaining low thermal thresholds during green extraction is critical to preserving the integrated polyphenolic architecture of B. sacra required for multi-targeted antimicrobial, anti-obesity, and anticancer therapeutics. Full article

Digestive enzymes such as pancreatic lipase (PL), α-glucosidase (AG), and α-amylase (AA) play key roles in the hydrolysis of dietary lipids and carbohydrates and are therefore relevant biochemical targets for evaluating compounds with potential to modulate postprandial metabolic responses. Neurolaena lobata (L.) R. Br. ex Cass. (Asteraceae) is a neotropical medicinal species traditionally used for several health-related conditions, including metabolic complaints. This study evaluated the in vitro inhibitory activity of the hydroalcoholic extract, selected VLC fractions, and isolated metabolites from N. lobata leaves against PL, AG, and AA. Enzyme inhibition screening of the extract and VLC fractions was used to select fractions for phytochemical investigation, affording six known metabolites: the sesquiterpene lactones neurolenin B (C1) and lobatin A (C2), the benzoic acid derivatives p-hydroxybenzoic acid (C3) and 3,4-dihydroxybenzoic acid (C4), and the flavonoids 6-hydroxykaempferol 3,7-dimethyl ether (C5) and quercetagetin 3,7-dimethyl ether (C6). The hydroalcoholic extract and the MeOAc and iPrOH fractions inhibited PL, AG, and AA. Among the isolated compounds, C1, C2, C5 and C6 inhibited PL and AG, with IC₅₀ values ranging from 134 to 615 µM and from 170 to 639 µM, respectively, whereas all compounds showed weak AA inhibition. Exploratory kinetic analysis yielded apparent inhibition profiles mainly consistent with competitive behavior; C6 showed an apparent profile consistent with noncompetitive inhibition against AG under the assay conditions used. These findings expand the phytochemical and in vitro bioactivity profile of N. lobata and provide preliminary evidence of digestive enzyme modulation in isolated enzyme systems, without establishing therapeutic efficacy. Full article

Dihydrocaffeic acid (DHCA) is a naturally occurring phenolic acid with recognized antioxidant and biological properties. However, its relatively high polarity limits its applicability in lipid-based systems. In this study, myrtenyl dihydrocaffeate was synthesized through lipase-catalyzed esterification of DHCA with myrtenol using immobilized Candida antarctica lipase B. The reaction conditions were optimized using response surface methodology based on a central composite design, yielding an experimental ester yield of 39.94 ± 1.16%. The synthesized ester was characterized by NMR spectroscopy and subsequently evaluated using a combination of experimental and in silico approaches. Antioxidant activity was determined by DPPH^• and ABTS^•+ radical scavenging assays, while oxidative stabilization of rapeseed oil was assessed by pressure differential scanning calorimetry (PDSC). Antimicrobial activity was evaluated using disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. In silico studies included ADMET profiling, PASS bioactivity prediction, protein target prediction, and molecular docking. These computational analyses were used only as hypothesis-generating tools because the predicted protein targets had low target-probability scores and were not experimentally validated. Myrtenyl dihydrocaffeate retained substantial antioxidant activity and significantly improved the oxidative stability of rapeseed oil, exhibiting protection factors comparable to those of DHCA. The ester also demonstrated mild antimicrobial activity against selected Gram-positive bacteria. Overall, the results indicate that lipophilization of DHCA with myrtenol is an effective strategy for developing lipophilic antioxidant derivatives for lipid-based food, cosmetic, or topical formulations, while the predicted molecular targets require experimental validation. Full article

Microwave-assisted enzymatic ring-opening polymerization (eROP) has emerged as a promising green approach for the synthesis of biomedical polymers with potential applications in drug delivery systems and implantable biomaterials. This review provides a critical overview of the current state of microwave-assisted eROP, with particular emphasis on systems catalyzed by Candida antarctica lipase B (CALB), the most widely used biocatalyst in this field. The fundamental aspects of CALB-catalyzed polymerization, including its catalytic mechanism and key parameters influencing reaction efficiency, are discussed under both conventional and microwave-assisted conditions. Microwave irradiation can offer significant advantages, such as reduced reaction times, improved energy efficiency, and, in some cases, enhanced polymer properties, including higher molecular weight and narrower dispersity. However, its effects are strongly dependent on reaction conditions, including solvent properties, temperature, and enzyme stability, with both accelerating and inhibiting effects reported. The review summarizes the current literature on microwave-assisted eROP of representative cyclic monomers, including ε-caprolactone, ω-pentadecalactone, and L-lactide, highlighting the influence of microwave parameters on polymerization outcomes and material properties. Despite promising results, microwave-assisted eROP remains relatively underexplored, with limited monomer scope and incomplete understanding of microwave–enzyme interactions. Challenges such as enzyme deactivation, lack of standardized methodologies, and scalability issues are also addressed. Overall, microwave-assisted eROP of cyclic monomers represents a sustainable and efficient strategy for producing high-quality pharmaceutical polymers. However, further research is required to fully realize its potential in industrial and biomedical applications. Full article

Entomopathogenic fungi of the genus Metarhizium can degrade and penetrate the insect cuticle through the coordinated action of hydrolytic enzymes, mainly lipases, proteases, and chitinases, whose production varies according to the fungal species and fermentation conditions. These enzymes can be generated via submerged fermentation and subsequently employed to enhance the insecticidal activity of fungal conidia. This study aimed to increase protease production from Metarhizium robertsii Mt015 to strengthen biological control agents based solely on fungal biomass. The culture medium composition and physicochemical parameters were optimized using a statistical design approach. Biological activity assays were then performed using Tuta absoluta larvae as the target insect and Beauveria bassiana as the reference control, tested both alone and in combination with the protease extract. Optimization identified wheat bran, casein, and an initial pH of 8–10 as the most influential variables, achieving a 5.5-fold increase in protease activity compared to the basal medium. When the protease extract was combined with B. bassiana conidia, the mortality rate reached 78.2%, significantly higher than the 55.6% achieved with B. bassiana conidia alone. Bliss independence analysis indicated that the observed larval mortality exceeded the additive expectation (Δ = 23.3 percentage points; 95% CI: 16.7–30.0), supporting a synergistic interaction between B. bassiana and the protease extract at 0.68 U/mL. These results demonstrate that enzymatic supplementation markedly improves the insecticidal performance of entomopathogenic fungi, supporting the use of enzyme-enriched formulations as a complementary strategy to strengthen biological control agents and advance the development of next-generation biopesticides. Full article

This study assessed the effects of adding slightly acidic electrolyzed water (SAEW) to drinking water (0.3 and 0.6 mg/L) for 15 consecutive days on the growth performance, digestive enzyme activity, amino acid transporter gene expression, antioxidant activity, and immune function in weaned piglets. Using a randomized complete block design with pen as the experimental unit, 144 healthy weaned piglets were randomly assigned to three groups with four replicate pens per group according to weight and sex. During the entire experimental period, the average daily gain and average daily feed intake of the SAEW groups were significantly higher than those of the control (CON) group (p < 0.05). As compared to the CON group, the levels of α-amylase, β-amylase, lipase, and proteases in the duodenum increased in both SAEW groups (p < 0.05). Furthermore, 0.6 mg/L SAEW positively affected the mRNA levels of CAT1, PepT1, CAT2, EAAC1, rBAT, b0,+ AT, and 4F2hc in the jejunum and duodenum (p < 0.05). Regarding immune function, SAEW significantly decreased intestinal secretory immunoglobulin A levels in a dose-dependent manner (p < 0.05), while no significant changes were observed in serum immunoglobulin concentrations between the SAEW groups and the CON group. In terms of antioxidant status, SAEW significantly increased superoxide dismutase and glutathione peroxidase activities, but also increased malondialdehyde content in a dose-dependent manner. Despite the stronger promotion of intestinal amino acid transporters under 0.6 mg/L SAEW, this advantage failed to generate extra growth benefits alongside elevated oxidative damage. In conclusion, both tested SAEW concentrations exert positive effects on piglet growth and digestion; comprehensively balancing production benefits and redox safety, 0.3 mg/L SAEW is the optimal drinking water dosage for weaned piglets. Full article

This study investigated the effects of dietary Bacillus subtilis supplementation on growth performance, digestive enzyme activity, expression of antioxidant- and inflammation-related genes, and gut microbiota in juvenile GIFT (Oreochromis niloticus). The juvenile GIFT (total 540, initial body weight: 16.17 ± 1.32 g) were randomly divided into six groups and fed diets containing 0, 1 × 10⁷, 1 × 10⁸, 1 × 10⁹, 1 × 10¹⁰, or 1 × 10¹¹ CFU/kg B. subtilis for 60 days. Compared with the control group (0 CFU/kg B. subtilis), dietary B. subtilis significantly improved final body weight, weight gain rate, specific growth rate, and daily growth index and reduced feed conversion ratio (p < 0.05). Supplementation also significantly increased intestinal amylase, lipase, and trypsin activities (p < 0.05). In addition, the expression of several antioxidant-related genes was upregulated, whereas pro-inflammatory gene expression was generally downregulated in the intestine, liver, and head kidney. Gut microbiota analysis indicated that B. subtilis supplementation altered community composition, increased Firmicutes’ abundance, and reduced Proteobacteria abundance. Among all supplementation levels evaluated, the diet containing 1 × 10⁹ CFU/kg B. subtilis produced the most favorable overall beneficial effects under the conditions of the present study. Full article

Severe acute pancreatitis (SAP) is a life-threatening inflammatory disorder characterized by high mortality and limited therapeutic options. Alginate oligosaccharide (AOS), a marine-derived bioactive polysaccharide, exhibits prebiotic, anti-inflammatory and antioxidant properties that are effective against various inflammatory diseases. In this study, a mouse model of SAP was established by intraperitoneal injection of cerulein (100 μg/kg) and lipopolysaccharide (5 mg/kg), and the mice were pretreated with AOS (200 mg/kg) by gavage for 4 consecutive weeks to explore the potential protective efficacy and underlying mechanisms. The results shown that AOS attenuated the severity of SAP, as evidenced by reduced serum amylase and lipase levels, as well as alleviated histopathological injury in both pancreatic and ileal tissues. AOS suppressed the overproduction of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) in serum, pancreas, and ileum at protein or mRNA levels. Moreover, AOS effectively diminished pancreatic and ileal inflammatory infiltration and oxidative stress in SAP mice, accompanied by inhibited the TLR4/MyD88/NF-κB pathway and activated the Nrf2/HO-1 antioxidant axis. Furthermore, AOS restored intestinal barrier integrity, as manifested by upregulated expression of tight junction proteins (claudin-1, occludin, ZO-1), reduced serum diamine oxidase, and decreased bacterial translocation from the gut to the pancreas. It was revealed by 16S rRNA sequencing that AOS ameliorated SAP-induced gut dysbiosis by restoring microbial diversity, normalizing the Firmicutes/Bacteroidetes ratio, enriching beneficial genera (Lactobacillus, Blautia), and enhancing cecal short-chain fatty acid (acetic, propionic, butyric acid) production. Collectively, our findings demonstrate that AOS exerts comprehensive protective effects against SAP through suppression of inflammatory signaling and oxidative stress, as well as restoring gut homeostasis. These results suggest that AOS may serve as a promising prebiotic-based nutritional strategy for the management of SAP. Full article

Several strategies can be employed for the identification of novel microbial lipases. Despite the increasing importance of metagenomics in bioprospecting, significant limitations in the expression of recombinant proteins, and lipases in particular, remain. Culture-based bioprospecting approaches are, therefore, still valuable. In this work, a collection of bacterial isolates, mainly of marine origin, was screened for lipase activity through a culture-based approach. Screening for lipolytic bacteria was performed in solid media containing olive oil emulsions and rhodamine B. Positive isolates were subsequently grown in liquid media, to confirm lipase production. Significant hydrolytic activity towards the triglyceride substrates tributyrin and triolein could be observed with the biomass produced, although no lipase activity could be detected in the culture supernatants. Six isolates presenting high activity were characterized as whole-cell biocatalysts, and all were found to be active at temperatures ranging between 25 and 65 °C, and at pH values between 6 and 10.5. Genomic analyses of two of these Gram-negative lipase-producing isolates revealed the presence of several hypothetical genes encoding for lipolytic enzymes, including outer cell-bound enzymes, predicted through the application of machine-learning tools. These natural isolates, containing cell-associated lipases, may therefore be of special interest for application as whole-cell biocatalysts. Full article

Poly(ε-caprolactone) (PCL) is a biodegradable aliphatic polyester with applications in many areas. Lipase B from Moesziomyces antarcticus (CALB, previously known as Candida antarctica lipase B) is a well-characterize enzyme capable of hydrolysing several polyesters. In this study, the codon-optimized gene encoding CALB was cloned into the yeast Yarrowia lipolytica to enhance its natural capabilities toward polyesters biodegradation. PCL films biodegradation was conducted directly in the medium using the engineered yeast at 28 °C. Process optimization employing baffled flasks significantly improved degradation efficiency and reduced time to 24 h. This study showed that the engineered yeast Y. lipolytica is a promising host for polyester biodegradation. Full article

Artisanal raw milk cheeses harbour complex microbial communities that drive cheese making and shape sensory quality. Previous work on Idiazabal cheese identified rennet as a major microbial source, although all reservoirs contributed to varying degrees. However, their impact in terms of enzyme-encoding genes related to technological quality of cheese remained unexplored. Building on that, this study draws on metagenome-assembled genomes (MAGs) from cheeses and dairy environments to comprehensively identify enzyme-encoding genes involved in key biochemical processes. In cheese MAGs (Lacticaseibacillus paracasei), protease-encoding genes were dominated by ATP-dependent metalloproteases (M41), carbohydrate-active enzyme-encoding genes (CAZymes) by glycoside hydrolases (GH) and glycosyltransferases (GT), while esterase, lipase, and related-enzyme-encoding genes were restricted to sparse ‘GDXG’, type-B and esterase D families. Dairy environments emerged as major reservoirs of enzyme-encoding genes, with notable differences among sample types (p ≤ 0.001). The richest sources of protease-encoding genes were grass (610 genes), linked primarily to Pantoea agglomerans, and rennet (318), mainly related to Basfia sp. and Moraxella sp., dominated by metalloproteases (M23, M38) and serine proteases (S15). The largest reservoirs of CAZyme-encoding genes were food contact surfaces (1550), associated mainly with Salinisphaera sp. and Dietzia sp., and rennet (1505), related to, e.g., Bacteroides pyogenes, Alloprevotella sp., and Lentilactobacillus buchneri. Food contact surfaces were also the richest source of esterase, lipase and related-enzyme-encoding genes (1209), mainly linked to Dietzia sp., Corynebacterium sp., and Brevibacterium aurantiacum. Similarly, aroma-related enzyme-encoding genes (e.g., oppA, pepA, GH13, esterase D) were consistently detected in environmental matrices. These results provide novel insights into dairy microbiomes as functional reservoirs of aroma precursors, revealing their relevance for artisanal PDO cheese production and future biotechnological applications. Full article

This study addressed the issue of insufficient activity in CALB lipase during the catalytic synthesis of key chiral intermediates for moxifloxacin. A structure-guided protein engineering strategy was employed to systematically modify its functional domains. Through molecular dynamics simulations of CALB-I189K, multiple regions exhibiting high conformational flexibility were preliminarily identified. Subsequently, by integrating 3D structural alignment with active site pocket distance analysis, the functionally most critical region (143–146) was selected. A site-directed saturation mutation library was constructed specifically targeting this region. Building upon the previously reported CALB-I189K, a mutant I189K/L144R/A146K was ultimately obtained through high-throughput screening combined with chiral HPLC validation. This mutant maintains excellent stereoselectivity (E = 206.52) while enhancing catalytic efficiency (k_cat/Κ_m) to 273.73 min⁻¹·mM⁻¹, approximately 4.5-fold that of I189K. At a substrate concentration of 1 M, it achieves 50% conversion within 2.6 h, demonstrating kinetic resolution capabilities approaching industrial standards. Molecular simulation analysis indicates that the L144R and A146K mutations synergistically enhance catalytic performance primarily by optimizing spatial distances between catalytic residues. This study not only provides a high-performance catalyst for the efficient biosynthesis of moxifloxacin chiral intermediates but also offers new insights for enzyme rational design based on dynamic structural information. Full article

This experiment investigated the effects of dietary Artemisia ordosica Krasch. (AOK) supplementation on the n3-polyunsaturated fatty acid (n3-PUFA) profile of subcutaneous adipose tissue (SADT) in Arbas cashmere goats and explored the underlying transcriptional mechanisms. Forty healthy, weaned kids (120 ± 10 days of age; similar body weight) were randomly allocated to two groups (n = 20): a control group (CON, basal diet) and an AOK group (AOK, basal diet with 3% of the roughage replaced by AOK). The feeding trial spanned 104 days, consisting of a 14-day adaptation period and 90 days of data acquisition. Compared with the CON group, AOK significantly reduced the content of saturated fatty acids (SFAs) and n6-polyunsaturated fatty acids (n6-PUFAs)/n3-PUFAs (n6/n3). In contrast, the levels of n3-PUFAs in the SADT of cashmere goats increased markedly (p < 0.05). Compared with the CON group, AOK exhibited significantly higher activities of hormone-sensitive lipase (HSL) (p = 0.027), adenylyl cyclase 2 (ADCY2) (p = 0.010), adenylyl cyclase 5 (ADCY5) (p = 0.046), cluster of differentiation 36 (CD36) (p = 0.013), solute carrier family 27 member 4 (SLC27A4) (p = 0.021), and fatty acid binding protein 4 (FABP4) (p = 0.040), along with significantly lower activities of fatty acid synthase (FAS) (p = 0.002), lipoprotein lipase (LPL) (p = 0.048), and stearoyl-coa desaturase (SCD) (p = 0.026) in SADT. Compared with the CON group, the activities of superoxide dismutase (SOD) (p = 0.032), catalase (CAT) (p = 0.010), glutathione peroxidase (GSH-PX) (p = 0.029), and total antioxidant capacity (T-AOC) (p = 0.002) were significantly increased in the AOK group. Transcriptomic profiling revealed that AOK supplementation downregulated mRNA levels of ADCY2, ADCY5, LPL, FAS, SCD, stearoyl-CoA desaturase 1 (SCD1), stearoyl-CoA desaturase 2 (SCD2), glycogen synthase 1 (GYS1), acyl-CoA oxidase 1 (ACOX1), acetyl-CoA carboxylase (ACC), diacylglycerol acyltransferase 1 (DGAT1), fatty acid desaturase 1 (FADS1), solute carrier family 27 member 2 (SLC27A2), erythroblastic leukemia viral oncogene homolog 4 (ERBB4), and carnitine palmitoyltransferase 1B (CPT1B) (p < 0.05). It also markedly induced acyl-CoA synthetase long-chain family member 4 (ACSL4) (p < 0.01) in SADT. Genes significantly enriched in the adenosine-monophosphate-activated protein kinase (AMPK) signaling pathway included LPL, SCD1, CPT1B, and GYS1 (p = 0.010). Genes significantly enriched in the phosphatidylinositol 3-kinase-akt (PI3K-Akt) signaling pathway included GYS1 and ERBB4 (p = 0.015). CPT1B, ADCY2, and GYS1 were identified as the genes significantly enriched in the insulin resistance signaling pathway (p = 0.048). LPL was the only gene significantly enriched in the cholesterol metabolism pathway (p = 0.049). Genes showing a tendency toward significant enrichment in the peroxisome-proliferator-activated receptor (PPAR) signaling pathway included ACSL4, CPT1B, SCD1, and LPL (p = 0.051). These interconnected cascades improve insulin sensitivity, stimulate triglyceride (TG) hydrolysis, and modulate n3-PUFA levels. Supplementation with AOK enhances n3-PUFA content by accelerating TG breakdown while simultaneously restraining FA oxidation in SADT. Consequently, AOK supplementation can be effectively used to enhance the nutritional value of cashmere goat meat through improved n3-PUFA deposition in SADT. Full article

Background and Clinical Significance: Burkitt lymphoma is an aggressive form of non-Hodgkin lymphoma of B-cell origin, caused by a MYC gene translocation on chromosome 8. There are three clinical subtypes, of which the sporadic subtype is most prevalent in the United States. Sporadic Burkitt lymphoma is diagnosed at a median age of 30 years and commonly manifests as bulky abdominal lesions, most often involving the ileocecal region. Pancreatic involvement is uncommon, and presentation as acute pancreatitis secondary to Burkitt lymphoma is exceedingly rare. Case Presentation: We present a case of a young male who presented with epigastric pain, nausea, and vomiting. He had a diffusely tender abdomen and elevated lipase levels. On imaging, he was found to have large retroperitoneal and intraperitoneal masses, contiguous with an enlarged pancreas. Burkitt lymphoma was confirmed upon biopsy of duodenal and gastric masses via endoscopic ultrasound. MRI brain and testicular ultrasound revealed unilateral fifth cranial nerve and bilateral testicular involvement, respectively. His course was complicated by bowel perforation requiring urgent surgery. However, he achieved complete remission with dose-dense systemic and intrathecal chemotherapy. Conclusions: This case highlights the diverse presentations of Burkitt’s lymphoma and a favorable prognosis with treatment. Clinicians should maintain a high index of suspicion for a malignant etiology of acute pancreatitis in patients without classic risk factors. Full article

This study evaluated activities of crude extracts from different parts of the tree of heaven (Ailanthus altissima (Mill.) Swingle) collected in Slovenia and Hungary, using effect-directed analyses based on hyphenation of high-performance thin-layer chromatography (HPTLC) and nine in vitro assays performed in situ on chromatographic plates after the separation. HPTLC separation combined with a set of four antibacterial assays, two antifungal assays, and three enzyme inhibitor assays to evaluate the extracts of 15 plant parts: young shoots, young leaves, mature leaves, yellow leaves, petioles of leaves, petioles of male inflorescences, petioles of fruits, female inflorescences, young fruits, male inflorescences, mature male inflorescences, bark of 1–2-year branches, bark of 2-year branches, bark of tree trunk, and bark of roots. Antibacterial activities against Gram-positive bacteria (Bacillus subtilis, Rhodococcus fascians) and Gram-negative bacteria (Aliivibrio fischeri, Pseudomonas syringae pv. maculicola (Psm)), as well as inhibition of enzymes α-glucosidase, lipase, and acetylcholinesterase, were observed for all extracts. Extracts differed in their antifungal activities. Extracts of young shoots, mature leaves, petioles of leaves, and bark of roots showed antifungal activity against plant pathogens Fusarium avenaceum and Bipolaris sorokiniana. Extracts of yellow leaves, male inflorescences, bark of 1–2-year branches, and bark of tree trunks were only active against F. avenaceum, whereas extracts of young leaves were only active against B. sorokiniana. This study is the first to report that A. altissima extracts exhibit (1) antifungal activity against F. avenaceum and B. sorokiniana; (2) antibacterial activity against A. fischeri, Psm, R. fascians, and B. subtilis (except leaves, bark of branches and bark of tree trunks); and (3) inhibitory activity toward lipase, α-glucosidase (except bark of tree trunks), and acetylcholinesterase (except bark of tree trunks). Full article