Search Results (104)

Hydrophobins are small, surface-active protein biosurfactants secreted by filamentous fungi with potential applications in industries such as pharmaceuticals, sanitation, and biomaterials. Additionally, hydrophobins are known to stabilize enzymatic processing of biomass for improved catalytic efficiency. In this study, Pichia pastoris was used to recombinantly express hydrophobin HFBI from Trichoderma reesei, a well-characterized fungal system used industrially for bioethanol production. Iterative optimization was performed on both the induction and purification of HFBI, ultimately producing yields of 86.6 mg/L HFBI and elution concentrations of 48 μM HFBI determined pure by SDS-PAGE, over a five-day methanol-fed batch shake flask induction regiment followed by a single unit operation multimodal cation exchange purification. This final purified material represents an improvement over prior approaches to enable a wider range of potential applications for biosurfactants. Full article

Concern over the presence of pharmaceutical waste in the environment has prompted research into the management of emerging organic micropollutants (EOMs). In response, sustainable technologies have been applied as alternatives to reduce the effects of these contaminants. This study investigated the capacity of filamentous fungi isolated from iron mine soil in the Amazon region to biodegrade the drug chloroquine diphosphate. An initial screening assessed the growth of four fungal strains on solid media containing chloroquine diphosphate: Trichoderma pseudoasperelloides CBMAI 2752, Penicillium rolfsii CBMAI 2753, Talaromyces verruculosus CBMAI 2754, and Penicillium sp. cf. guaibinense CBMAI 2758. Among them, Penicillium sp. cf. guaibinense CBMAI 2758 was selected for further testing in liquid media. A Box–Behnken factorial design was applied with three variables, pH (5, 7, and 9), incubation time (5, 10, and 15 days), and chloroquine diphosphate concentration (50, 75, and 100 mg·L⁻¹), totaling 15 experiments. The samples were analyzed by gas chromatography–mass spectrometry (GC-MS). The most effective conditions for chloroquine biodegradation were pH 7, 100 mg·L⁻¹ concentration, and 10 days of incubation. Four metabolites were identified: one resulting from N-deethylation M1 (N4-(7-chloroquinolin-4-yl)-N1-ethylpentane-1,4-diamine), two from carbon–carbon bond cleavage M2 (7-chloro-N-ethylquinolin-4-amine) and M3 (N1,N1-diethylpentane-1,4-diamine), and one from aromatic deamination M4 (N1-ethylbutane-1,4-diamine) by enzymatic reactions. The toxicity analysis showed that the products obtained from the biodegradation of chloroquine were less toxic than the commercial formulation of this compound. These findings highlight the biotechnological potential of Amazonian fungi for drug biodegradation and decontamination. Full article

Monascus purpureus is a filamentous fungus renowned for producing bioactive secondary metabolites, including lovastatin and azaphilone pigments. Lovastatin is valued for its cholesterol-lowering properties and cardiovascular benefits, while Monascus pigments exhibit anti-cancer, anti-inflammatory, and antimicrobial activities, underscoring their pharmaceutical and biotechnological relevance. This study evaluated the impact of carbohydrate-derived elicitors—mannan oligosaccharides, oligoguluronate, and oligomannuronate—on the enhancement of pigment and lovastatin production in M. purpureus C322 under submerged fermentation. Elicitors were added at 48 h in shake flasks and 24 h in 2.5 L stirred-tank fermenters. All treatments increased the production of yellow, orange, and red pigments and lovastatin compared to the control, with higher titres upon scale-up. OG led to the highest orange pigment yield (1.2 AU/g CDW in flasks; 1.67 AU/g CDW in fermenters), representing 2.3- and 3.0-fold increases. OM yielded the highest yellow and red pigments (1.24 and 1.35 AU/g CDW in flasks; 1.58 and 1.80 AU/g CDW in fermenters) and the highest lovastatin levels (10.46 and 12.6 mg/g CDW), corresponding to 2.03–3.03-fold improvements. These results highlight the potential of carbohydrate elicitors to stimulate metabolite biosynthesis and facilitate scalable optimisation of fungal fermentation. Full article

Tannase, a highly adaptive biocatalyst, plays a pivotal role in diverse bioconversion reactions in nature. This enzyme exhibits numerous applications across various industrial sectors, including food, pharmaceuticals, chemicals, and beverages. This study aimed to screen and characterize fungal endophytes isolated from mangrove plants for their enzyme tannase-producing ability. Eighty-five filamentous endophytic fungi were isolated from different mangrove samples and subsequently identified. These fungal strains were initially screened using the tannic acid agar plate method. Out of the screened strains, 13 fungal isolates demonstrated tannase production ability. The quantitative estimation of extracellular tannase was performed using the submerged fermentation technique. Among the studied endophytes, eight isolates, namely LV_084 (21.21 IU/mL), LV_074 (15.41 IU/mL), LV_078 (6.98 IU/mL), LV_038 (6.97 IU/mL), LV_077 (6.32 IU/mL), LV_016 and LV_066 (6.37 IU/mL), and LV_060 (6.18 IU/mL) exhibited excellent tannase activity. Among these isolates, LV_084 Phyllosticta capitalensis and LV_074 Aspergillus chevalieri showed the highest enzyme-producing ability. These isolates were authenticated using ITS rDNA sequencing, followed by BLAST search and phylogenetic analysis. Furthermore, the physical and chemical conditions for the maximum enzyme production were optimized. This is the first report of enzyme tannase production by Phyllosticta capitalensis and Aspergillus chevalieri. Full article

The valorization of cheese whey, a rich by-product of the dairy industry that is rich in lactose (approx. 70%), proteins (14%), and minerals (9%), represents a promising approach for microbial fermentation. With global whey production exceeding 200 million tons annually, the high biochemical oxygen demand underlines the important need for sustainable processing alternatives. This review explores the biotechnological potential of whey as a fermentation medium by examining its chemical composition, microbial interactions, and ability to support the synthesis of valuable metabolites. Functional microorganisms such as lactic acid bacteria (Lactobacillus helveticus, L. acidophilus), yeasts (Kluyveromyces marxianus), actinobacteria, and filamentous fungi (Aspergillus oryzae) have demonstrated the ability to efficiently convert whey into a wide range of bioactive compounds, including organic acids, exopolysaccharides (EPSs), bacteriocins, enzymes, and peptides. To enhance microbial growth and metabolite production, whey fermentation can be carried out using various techniques, including batch, fed-batch, continuous and immobilized cell fermentation, and membrane bioreactors. These bioprocessing methods improve substrate utilization and metabolite yields, contributing to the efficient utilization of whey. These bioactive compounds have diverse applications in food, pharmaceuticals, agriculture, and biofuels and strengthen the role of whey as a sustainable biotechnological resource. Patents and clinical studies confirm the diverse bioactivities of whey-derived metabolites and their industrial potential. Whey peptides provide antihypertensive, antioxidant, immunomodulatory, and antimicrobial benefits, while bacteriocins and EPSs act as natural preservatives in foods and pharmaceuticals. Also, organic acids such as lactic acid and propionic acid act as biopreservatives that improve food safety and provide health-promoting formulations. These results emphasize whey’s significant industrial relevance as a sustainable, cost-efficient substrate for the production of high-quality bioactive compounds in the food, pharmaceutical, agricultural, and bioenergy sectors. Full article

Phycobiliproteins and pigments derived from cyanobacteria hold significant potential for diverse applications in the food, pharmaceutical, and chemical industries. The filamentous cyanobacterium Anabaena cylindrica serves as a valuable resource for extracting these compounds. This study develops a simplified, safe, and cost-effective extraction method that eliminates toxic solvents and minimizes processing steps. This makes the method applicable for all users and allows the easy integration of the extraction into biorefinery concepts in which the biomass is to be used as a fertilizer, for example. Utilizing salts such as ammonium sulfate and calcium chloride (15 gL⁻¹ each) enables the effective extraction of phycocyanin (PC) and allophycocyanin, achieving a PC concentration of 192.34 $\mathrm{mg}{\mathrm{g}}_{\mathrm{CDW}}^{-1}$ and 209.44 $\mathrm{mg}{\mathrm{g}}_{\mathrm{CDW}}^{-1}$, respectively. Ethanol was introduced as a less toxic alternative to methanol for pigment extraction, increasing chlorophyll a and carotenoid recovery by 21% and 37%, respectively. Full article

Arthrospira platensis is a filamentous cyanobacterium produced commercially for human consumption, and it is a source of phycocyanin (PC), which recently stirred up great interest due to its anti-inflammatory, radical scavenging, antioxidant and hepato-protective properties. This work has studied the encapsulation of A. platensis extract in starch sodium octenyl succinate by employing freeze-drying and two spray-drying techniques, conventional and nanospray-drying. The main characteristics and properties, including PC encapsulation efficiency, size, colour, and thermal stability of the capsules, were evaluated. Moreover, the antioxidant capacity of encapsulated extract and release of PCs into saliva simulant, were studied and compared. Similar PC encapsulation efficiency was achieved using freeze-drying and nanospray-drying techniques with values of 67–71% and 70–78%, respectively. Meanwhile, the conventional spray-drying method achieved significantly lower encapsulation efficiency values (38–42%). The thermal stability of encapsulated A. platensis extract was improved as demonstrated by the higher decomposition temperature, which was increased by 8–11 °C, 11–15 °C, and 22–23 °C for spray-dried, nanospray-dried and freeze-dried samples, respectively. The nanospray-drying technique allowed the production of the smallest particles with an average diameter of 2–14 µm, good colour and thermal stability, and antioxidant capacity. Overall, the results demonstrated the potential of A. platensis extract encapsulation in modified starch using several techniques with potential application as bioactive ingredients in nutraceutical or pharmaceutical products. Full article

Background/Objectives: Chagas disease is a neglected tropical disease caused by infection with the parasite Trypanosoma cruzi. Benznidazole and nifurtimox are the only approved drugs for treating this condition, but their low aqueous solubility may lead to erratic bioavailability. This work aimed for the first time to formulate tablets of nifurtimox by hot melt extrusion coupled with 3D printing as a strategy to increase drug dissolution and the production of tablets with dosage on demand. Methods: Different pharmaceutical-grade polymers were evaluated through film casting, and those with promising nifurtimox amorphization capacity were further used to prepare filaments by hot melt extrusion. The printability of the obtained filaments was tested, and the polyvinyl alcohol filament was further used for printing tablets containing 120 and 60 mg of nifurtimox. Results: Three-dimensional tablets showed a remarkable improvement in the drug dissolution rate compared to commercial tablets and a dissolution efficiency 2.8 times higher. In vivo studies were carried out on Swiss mice. Parasitemia curves of nifurtimox printed tablets were significantly superior to the pure drug. Moreover, NFX 3D tablets provided a similar Trypanosoma cruzi reduction in plasmatic concentration to benznidazole, the gold-standard drug for acute-phase treatment of the Chagas disease. Conclusions: The findings of this work showed that hot melt extrusion coupled with 3D printing is a promising alternative for increasing nifurtimox biopharmaceutical properties and an attractive approach for personalized medicine. Full article

Phase separation is quite common in formulations for hydrophobic active pharmaceutical ingredients (APIs) due to their thermodynamic instability in a supersaturated state during in vitro dissolution or in vivo absorption. Phase separation possibly accompanies the formation of a disordered drug-rich phase, but this is still not thoroughly understood. In this study, the phase separation of supersaturated axitinib (Axi) in media with or without polymers was evaluated via multiple analytical methods, including UV–vis and fluorescence spectroscopy, dynamic light scattering, and microscopy. The phase separation of Axi occurred at an Axi concentration of 25–30 µg/mL in the media, while the addition of quantitative hypromellose acetate succinate (HPMCAS) MG and povidone (PVP) K30 did not alter its phase separation concentration. The second scattering dispersion phase of the system exhibited superior stability and reversibility as the formative filamentous crystalline condensates could disintegrate upon dilution. These disparate analyses consistently detected the phase separation of Axi. This manuscript could provide a better understanding of the supersaturation state of hydrophobic APIs upon pharmaceutical application. Full article

Fast-growing mycobacteria cause difficult-to-treat infections due to their high intrinsic resistance to antibiotics as well as disinfectant agents. Mycobacterium abscessus complex (MAC) is the main cause of nontuberculous mycobacteria diseases. In this work, we evaluated the activity of the novel synthetic antimicrobial peptide, Agelaia-12, against Mycobacterium abscessus and M. massiliense. Agelaia-12 showed a minimum inhibitory concentration (MIC) of 25 μM detected against M. abscessus and M. massiliense with no cytotoxicity. The scanning electronic microscopy analysis of mycobacterial treated with Agelaia-12 demonstrated the presence of filamentous structures and aggregation of the cells. Congo red binding assay of M. abscessus exhibited altered dye accumulation after treatment with Agelaia-12. Treatment of M. abscessus- or M. massiliense-infected murine macrophages with Agelaia-12 decreased the mycobacterial load by 92% for the tested strains. Additionally, IFN-y KO mice infected with M. abscessus or M. massiliense and treated with Agelaia-12 showed a 98% reduction in lung bacterial load. Thus, the synthetic peptide Agelaia-12 may be a promising biomolecule for the treatment of mycobacteriosis, and its structural properties may serve as a foundational model for the design and development of novel pharmaceutical agents aimed at combating this disease. Full article

Background/Objectives: Three-dimensional printing technology has emerging interest in pharmaceutical manufacturing, offering new opportunities for personalized medicine and customized drug delivery systems. Fused deposition modeling (FDM) is highly regarded in the pharmaceutical industry because of its cost effectiveness, easy operation, and versatility in creating pharmaceutical dosage forms. This review investigates different methods of incorporating active pharmaceutical ingredients (APIs) into filament matrices for use in fused deposition modeling (FDM) 3D printing. Methods: Two electronic databases, the Web of Science and PubMed, were utilized to survey the literature. The selected keywords for this review were as follows: fused filament fabrication OR fused deposition modeling OR FDM OR FFF AND 3D printing AND loading techniques OR impregnation techniques AND solid dosage form. Results: This paper evaluates various loading techniques such as soaking, supercritical impregnation, microwave impregnation, and hot-melt extrusion, focusing on their effectiveness and capacity for drug incorporation. Additionally, this review includes a thorough risk assessment of the extrusion process using Ishikawa and SWOT analyses. Conclusions: Overall, this review provides comprehensive insights into the latest advancements in 3D printing for pharmaceutical applications and identifies key areas for future research and development. Full article

The metabolic activities of microorganisms to modify the chemical structures of organic compounds are an effective tool for the production of high-value steroidal drugs or active pharmaceutical ingredients (APIs). The integration of biotransformation into the synthesis of APIs can greatly reduce the number of reaction steps and achieve higher process efficiency, thus enabling their greener production. The current research efforts are focused on either the optimization of existing processes or identification of new potentially useful bioconversions. This study aimed to assess the catalytic abilities of the filamentous fungus Fusarium culmorum AM 282 to transform B-nor analogues (5(6→7)abeo compounds) of steroid hormones: androstenedione (AD), dehydroepiandrosterone (DHEA) and its acetate. Our previous studies have demonstrated that this strain is an active hydroxylating catalyst for many steroidal compounds with diverse structures. The results presented in this work showed that the hydroxylation of B-nor steroids occurred with the regio- and stereoselectivity typical of this strain in relation to the corresponding natural hormones of the standard 6:6 A/B series. After the transformations of B-nor-DHEA and its acetate, 15α-hydroxy-B-nor-DHEA was obtained as the sole product of the reaction, while the transformation of the AD analogue resulted in a mixture of its 15α- and 6α-hydroxy derivatives. A detailed analysis of the transformation course indicated that all the obtained hydroxy derivatives could be the result of the activity of the same enzyme. The presented results may provide a basis for research aimed at understanding the molecular nature of cytochrome P-450 monooxygenase from F. culmorum AM 282 with its ability for 15α-hydroxylation of steroidal compounds. An analysis of the pharmacokinetic and pharmacodynamic properties of the obtained metabolites with cheminformatics tools suggests their potential biological activity. Full article

Tyrosinase is an important enzyme in the biosynthesis of melanin. Many skin-whitening agents that inhibit tyrosinase activity from natural sources have been identified because they are harmless and non-toxic. In this work, 114 samples of 54 Fabaceae plants were assessed for their anti-tyrosinase activity using a dopachrome method. The results found that Desmodium heterocarpon stems and roots demonstrated the highest tyrosinase inhibitory activity at 20 µg/mL (92.50 ± 1.09%), whereas the water extract of Artocarpus lacucha and kojic acid demonstrated 87.41 ± 0.61% and 95.71 ± 0.33%, respectively. Six compounds were isolated from this plant, including genistein (1); hexadecanoic acid (2); salicylic acid (3); β-sitosterol-D-glucoside (4); 2,3-dihydroxybenzoic acid (5); and 2,5-dihydroxybenzoic acid (6). Among them, 2,5-dihydroxybenzoic acid demonstrated a potential effect for tyrosinase inhibition with an IC₅₀ of 57.38 µg/mL, while standards of kojic acid and the water extract of A. lacucha showed 2.46 and 0.15 µg/mL, respectively. 2,3-dihydroxybenzoic acid had a similar structure as 2,5-dihydroxybenzoic acid; however, it was shown to have tyrosinase inhibitory activity, with an IC₅₀ of 128.89 µg/mL. Studies using computer simulations confirmed this reservation. The determination of antimicrobial activities found that 2,5-dihydroxybenzoic acid showed the strongest inhibitory activity against Staphylococcus aureus, with MIC and MBC of 5 and 5 µg/mL, respectively. In addition, it inhibited MRSA, S. epidermidis, Propionibacterium acnes, Escherichia coli, and Pseudomonas aeruginosa, with MIC and MBC of 15–30 and 15–40 µg/mL. It showed potential activities against yeast and filamentous fungi, such as Candida albicans, Microsporum gypseum, Trichophyton rubrum, and T. mentagrophytes, with MIC and MFC of 15 µg/mL. So, 2,5-dihydroxybenzoic acid could inhibit tyrosinase activity and microorganisms that cause skin diseases. Therefore, it can be concluded that this plant has advantageous properties that will be investigated and further developed for possible uses, particularly in the cosmetic and pharmaceutical industries. Full article

Additive manufacturing of pharmaceutical formulations offers advanced micro-structure control of oral solid dose (OSD) forms targeting not only customised dosing of an active pharmaceutical ingredient (API) but also custom-made drug release profiles. Traditionally, material extrusion 3D printing manufacturing was performed in a two-step manufacturing process via an intermediate feedstock filament. This process was often limited in the material space due to unsuitable (brittle) material properties, which required additional time to develop complex formulations to overcome. The objective of this study was to develop an additive manufacturing MicroFactory process to produce an immediate release (IR) OSD form containing 250 mg of mefenamic acid (MFA) with consistent drug release. In this study, we present a single-step additive manufacturing process employing a novel, filament-free melt extrusion 3D printer, the MicroFactory, to successfully print a previously ‘non-printable’ brittle Soluplus^®-based formulation of MFA, resulting in targeted IR dissolution profiles. The physico-chemical properties of 3D printed MFA-Soluplus^®-D-sorbitol formulation was characterised by thermal analysis, Fourier Transform Infrared spectroscopy (FTIR), and X-ray Diffraction Powder (XRPD) analysis, confirming the crystalline state of mefenamic acid as polymorphic form I. Oscillatory temperature and frequency rheology sweeps were related to the processability of the formulation in the MicroFactory. 3D printed, micro-structure controlled, OSDs showed good uniformity of mass and content and exhibited an IR profile with good consistency. Fitting a mathematical model to the dissolution data correlated rate parameters and release exponents with tablet porosity. This study illustrates how additive manufacturing via melt extrusion using this MicroFactory not only streamlines the manufacturing process (one-step vs. two-step) but also enables the processing of (brittle) pharmaceutical immediate-release polymers/polymer formulations, improving and facilitating targeted in vitro drug dissolution profiles. Full article

Fiber-based technologies are widely used in various industries, but their use in pharmaceuticals remains limited. While melt extrusion is a standard method for producing medical fibers such as sutures, it is rarely used for pharmaceutical fiber-based dosage forms. The EsoCap system is a notable exception, using a melt-extruded water-soluble filament as the drug release trigger mechanism. The challenge of producing drug-loaded fibers, particularly due to the use of spinning oils, and the processing of the fibers are addressed in this work using other approaches. The aim of this study was to develop processes for the production and processing of pharmaceutical fibers for targeted drug delivery. Fibers loaded with polyvinyl alcohol and fluorescein sodium as a model drug were successfully prepared by a continuous melt extrusion process and directly spun. These fibers exhibited uniform surface smoothness and consistent tensile strength. In addition, the fibers were further processed into tubular dosage forms using a modified knitting machine and demonstrated rapid drug release in a flow cell. Full article