Search Results (28)

Cyanobacteria are a prolific source of bioactive metabolites with expanding applications in sustainable agriculture and biotechnology. This work explores, for the first time in thermotolerant Colombian isolates, the impact of light spectrum, photoperiod, and irradiance on the co-production of exopolysaccharides (EPS) and indole-3-acetic acid (IAA). Six strains from hot-spring environments were screened under varying blue:red (B:R) LED ratios and full-spectrum illumination. Hapalosiphon sp. UFPS_002 outperformed all others, reaching ~290 mg L⁻¹ EPS and 28 µg mL⁻¹ IAA in the initial screen. Response-surface methodology was then used to optimize light intensity and photoperiod. EPS peaked at 281.4 mg L⁻¹ under a B:R ratio of 1:5 LED, 85 µmol m⁻² s⁻¹, and a 14.5 h light cycle, whereas IAA was maximized at 34.4 µg mL⁻¹ under cool-white LEDs at a similar irradiance. The quadratic models exhibited excellent predictive power (R² > 0.98) and a non-significant lack of fit, confirming the light regime as the dominant driver of metabolite yield. These results demonstrate that precise photonic tuning can selectively steer carbon flux toward either EPS or IAA, providing an energy-efficient strategy to upscale thermotolerant cyanobacteria for climate-resilient biofertilizers, bioplastics precursors, and other high-value bioproducts. Full article

This study explores the valorization of agro-industrial by-products—riceberry broken rice (RBR) and soybean meal (SBM)—as cost-effective substrates for enhancing exopolysaccharide (EPS) production by Bacillus tequilensis PS21. Eight Bacillus strains were screened, and B. tequilensis PS21 demonstrated the highest EPS yield (2.54 g/100 mL DW). The EPS displayed a strong antioxidant capacity with 65.5% DPPH and 80.5% hydroxyl radical scavenging, and a FRAP value of 6.51 mg Fe²⁺/g DW. Antimicrobial testing showed inhibition zones up to 10.07 mm against Streptococcus agalactiae and 7.83 mm against Staphylococcus aureus. Optimization using central composite design (CCD) and the response surface methodology (RSM) revealed the best production at 5% (w/v) RBR, 3% (w/v) SBM, pH 6.66, and 39.51 °C, yielding 39.82 g/L EPS. This EPS is a moderate-molecular-weight (11,282 Da) homopolysaccharide with glucose monomers. X-ray diffraction (XRD) showed an amorphous pattern, favorable for solubility in biological applications. Thermogravimetric analysis (TGA) demonstrated thermal stability up to ~250 °C, supporting its suitability for high-temperature processing. EPS also exhibited anticancer activity with IC₅₀ values of 226.60 µg/mL (MCF-7) and 224.30 µg/mL (HeLa) at 72 h, reduced colony formation, inhibited cell migration, and demonstrated anti-tyrosinase, anti-collagenase, and anti-elastase effects. This study demonstrates the successful valorization of agro-industrial by-products—RBR and SBM—for the high-yield production of multifunctional EPS with potent antioxidant, antimicrobial, and anticancer properties. The findings highlight the sustainable potential of these low-cost substrates in supporting the development of green and value-added bioproducts, with promising utilizations across the food, pharmaceutical, and cosmetic sectors. Full article

In the realm of sustainable and eco-friendly agriculture, current scientific research emphasizes the development of plant-based bioproducts to mitigate the agricultural footprint resulting from excessive fertilizer and pesticide use. This study investigates the cladodes of Opuntia ficus-indica to screen for bioactive compounds and assess their efficacy against fungal pathogens isolated from infected tomato fruits. Quantitative analysis of the methanolic extract revealed substantial concentrations of bioactive compounds: total polyphenols (86.6 mg GAEs/100 g FW), flavonoids (13.4 mg QEs/100 g FW), condensed tannins (8.9 mg TAEs/100 g FW), and carotenoids (0.9 mg β-CEs/100 g FW). Notably, the DPPH assay indicated that the cladode extract exhibited significant antioxidant potential at a concentration of 0.6 mg/mL. Seven fungal pathogens were isolated from infected tomato fruits and identified as belonging to the following genera: Rhizoctonia (EC2), Fusarium (EC1 and EC3), Alternaria (EC4), Mucor (EC5), Aspergillus (EC6), and Penicillium (EC7). At a concentration of 0.02% of the cladode hydroethanolic extract, the antifungal activity results demonstrated mycelial growth inhibition for Alternaria sp. (70.91%), Rhizoctonia solani EC2 (58.49%), Fusarium oxysporum EC3 (57.63%), and Fusarium solani EC1 (53.13%). Conversely, lower inhibitory activities were observed for Mucor sp. EC5 (31.08%), Aspergillus sp. EC6 (35.14%), and Penicillium sp. EC7 (28.38%). At a concentration of 0.04%, all cladode hydroethanolic extracts inhibited mycelial growth by more than 50%. Furthermore, the highest spore inhibition was attained with the 0.04% cladode hydroethanolic extract (exceeding 50%). Inhibition percentages of 83.02%, 85.96%, 87.76%, and 90.20% were recorded for Fusarium oxysporum EC3, Fusarium solani EC1, Rhizoctonia solani EC2, and Alternaria sp. EC4, respectively. Collectively, these findings suggest that Opuntia ficus-indica extract holds significant promise for application as a biopesticide against fungal pathogens affecting tomato fruits. Full article

Methanotrophs hold significant potential in global methane mitigation and resource recovery. However, the limited rate of cell proliferation remains a significant constraint for large-scale applications. Therefore, screening efficient methanotrophic strains that are suitable for industrial applications to mitigate methane and exploring potential methane resource utilization pathways are of great importance for sustainable development. Gradient dilution and the streak plate method were employed to isolate methanotrophic strains from a previously domesticated methane-oxidizing microbial consortium. We isolated a highly efficient strain, M6, which exhibited a 230% increase in growth rate compared to the laboratory model strain Methylocystis bryophila (M. bryophila). Taxonomic analysis revealed that strain M6 is classified as Methylocystis parvus. Genomic data indicated a diverse range of metabolic functions. In addition to utilizing methane, strain M6 can also utilize citrate to generate energy and intermediate products, addressing issues related to insufficient methane supply or low methane mass transfer efficiency. Metabolic adaptability ensures the stability of its application. The optimal cultivation conditions for strain M6 were determined, characterized by mild and easily implementable parameters. Based on the analysis of the genome and metabolic pathways, strain M6 exhibits potential for the synthesis of bioproducts, such as proteins, lipids, and polyhydroxyalkanoates (PHAs), with the fermentation process not requiring cost-intensive carbon sources, making it both economical and sustainable. Full article

Transcription factor-based biosensors (TFBs) are powerful tools in microbial biosensor applications, enabling dynamic control of metabolic pathways, real-time monitoring of intracellular metabolites, and high-throughput screening (HTS) for strain engineering. These systems use transcription factors (TFs) to convert metabolite concentrations into quantifiable outputs, enabling precise regulation of metabolic fluxes and biosynthetic efficiency in microbial cell factories. Recent advancements in TFB, including improved sensitivity, specificity, and dynamic range, have broadened their applications in synthetic biology and industrial biotechnology. Computational tools such as Cello have further revolutionized TFB design, enabling in silico optimization and construction of complex genetic circuits for integrating multiple signals and achieving precise gene regulation. This review explores innovations in TFB systems for microbial biosensors, their role in metabolic engineering and adaptive evolution, and their future integration with artificial intelligence and advanced screening technologies to overcome critical challenges in synthetic biology and industrial bioproduction. Full article

The rapid growth of the human population has significantly increased the demand for food, leading to the intensification of agricultural practices that negatively impact the environment. Climate change poses a significant threat to global food production, as it can disrupt crop yields and modify the lifecycle stages of phytopathogens and pests. To address these challenges, the use of microbial inoculants, which are bioproducts containing beneficial microorganisms known as plant growth promotion microorganisms (PGPMs), has emerged as an innovative approach in sustainable agriculture. This review covers the isolation and identification of beneficial strains, the screening and selection process, the optimization of production techniques, and the importance of quality control and field testing. It also discusses the key points for the development and formulation of high-quality microbial inoculants, as well as highlights their advancements, current challenges, and future directions for research and application. Full article

The management of the organic fraction of municipal solid waste (OFMSW), also called urban biowaste, and urban wastewater sludge (UWWS) represents a challenge for cities and regions, which want to adopt innovative urban bioeconomy approaches for their treatment and production of high-added-value products beyond the traditional anaerobic digestion (AD) and compost. This adoption is often restricted by the availability and maturity of technologies. The research object of this manuscript, based on the findings of EU Horizon 2020 project HOOP, is the identification of state-of-the-art circular technologies for material valorisation of OFMSW and UWWS, following a novel screening methodology based on the scale of implementation (tested at least at pilot scale). The screening resulted in 25 technologies, which have been compared and discussed under a multidisciplinary assessment approach, showing their enabling factors and challenges, their current or potential commercial status and their compatibility with the traditional technologies for urban biowaste treatment (composting and AD). The bioproducts cover market sectors such as agriculture, chemistry, nutrition, bioplastics, materials or cosmetics. Therefore, the results of this review help project promoters at city/region level to select innovative technologies for the conversion of OFMWS and UWWS into high value products. Full article

Melanin is a high value bioproduct generated through the fermentation of Aureobasidium pullulans, playing a crucial role in various fields, including food, medicine, environmental protection, and materials science. However, its high production costs and low synthetic yields significantly limit its applications. Therefore, it is essential to identify high-yield strains, reduce production costs, and optimize fermentation strategies. In this study, a high melanin-yielding Aureobasidium pullulans 53LC7 was screened and identified, and the fermentation process was optimized based on melanin yield, color value, and pullulan yield. The results indicated that the melanin yield peaked at an initial pH of 6.0, temperature of 27 °C, fermentation time of 6.5 d, and inoculation quantity of 2.5%, achieving a melanin yield of 16.33 g/L. Subsequently, huangjiu lees, a byproduct of huangjiu production, was incorporated into the fermentation medium, resulting in a melanin yield of 5.91 g/L. This suggests that the Aureobasidium pullulans was not effectively utilizing huangjiu lees. To address this, we employed an adaptive evolution strategy, which increased the melanin yield to 8.72 g/L. The enhanced production was correlated with the expression of key genes, including FKS, PKS, and Cmr1. Finally, cellulase was utilized to convert the crude fibers in huangjiu lees, which were difficult to utilize, into usable substrates, while pullulanase was employed to minimize byproduct formation in the fermentation system, resulting in a melanin yield of 19.07 g/L. This study not only provides promising strains for further research but also offers valuable insights for resource production technologies. Full article

Living in diverse environmentally harsh conditions, the plant exhibits a unique survival mechanism. As a result, the endophytes residing within the plant produce specific compounds that promote the plant’s growth and defend it against pathogens. Plants and algae symbiotically harbor endophytes, i.e., microbes and microorganisms living within them. The objective of this study is to isolate endophytic fungi, specifically strains of Aspergillus terreus, from the leaves of the salt-tolerant plant Tetraena qatarensis and to explore the salt tolerance, antagonistic activity, and growth promotion properties. Strain C A. terreus (ON117337.1) was screened for salt tolerance and antagonistic effects. Regarding salt tolerance, the isolate demonstrated the ability to thrive in a concentration of up to 10% NaCl. A. terreus showed inhibitory activity against four fungal phytopathogens, namely Fusarium oxysporum, Alternaria alternata, Colletotrichum gloeosporioides, and Botrytis cinerea. The GC-MS investigation of the fungal (strain C Aspergillus terreus) extract showed the presence of about 66 compounds (secondary metabolites). Secondary metabolites (SMs) are produced, like Hexadecanoic acid, which aids in controlling phytopathogens. Also produced is lovastatin, which is used to treat hypercholesterolemia. Strain C, which showed salinity tolerance and the highest inhibitory activity, was further analyzed for its effect on tomato seed germination under pathogen stress from Fusarium oxysporum. The greenhouse experiment indicated that the fungi increased the length of tomato seedlings and the plant biomass. Therefore, the selected endophytes derived from Tetraena qatarensis were scrutinized for their potential as biocontrol agents, aiming to thwart fungal pathogens and stimulate plant growth. The in vitro and in vivo assessments of strain C (Aspergillus terreus) against Fusarium oxysporum in this investigation indicate the promising role of endophytes as effective biological control agents. Investigating novel bio-products offers a sustainable approach to agriculture, gradually reducing dependence on chemical fungicides. Full article

Carotenoids are valuable pigments naturally occurring in all photosynthetic plants and microalgae as well as in selected fungi, bacteria, and archaea. Green microalgae developed a complex carotenoid profile suitable for efficient light harvesting and light protection and harbor great capacity for carotenoid production through the substantial power of the endogenous 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway. Previous works established successful genome editing and induced significant changes in the cellular carotenoid content in Chlamydomonas reinhardtii. This study employs a tailored carotenoid pathway for engineered bioproduction of the valuable ketocarotenoid astaxanthin. Functional knockout of lycopene ε-cyclase (LCYE) and non-homologous end joining (NHEJ)-based integration of donor DNA at the target site inhibit the accumulation of α-carotene and consequently lutein and loroxanthin, abundant carotenoids in C. reinhardtii without changes in cellular fitness. PCR-based screening indicated that 4 of 96 regenerated candidate lines carried (partial) integrations of donor DNA and increased ß-carotene as well as derived carotenoid contents. Iterative overexpression of CrBKT, PacrtB, and CrCHYB resulted in a 2.3-fold increase in astaxanthin accumulation in mutant ΔLCYE#3 (1.8 mg/L) compared to the parental strain UVM4, which demonstrates the potential of genome editing for the design of a green cell factory for astaxanthin bioproduction. Full article

This study addresses the production enhancement of pullulan, an extracellular polysaccharide with various applications. Pullulan is primarily produced by Aureobasidium pullulans (A. pullulans), and genetic modification is commonly used to increase its yield. However, there is a need for a more efficient and safer method. To achieve this, we designed a high-throughput screening system utilizing a unique fluorescent protein specific to pullulan. Ultraviolet (UV) mutagenesis was applied to create a pool of mutant strains, and flow cytometry allowed for single-cell screening. Our approach yielded strain M1-B3, which exhibited a substantial increase in pullulan production from 26.5 g/L to 76.88 g/L. Additionally, the molecular weight of the produced pullulan significantly increased, expanding its potential commercial application. This study demonstrates an efficient and safe method to enhance pullulan production in A. pullulans. The UV mutagenesis and flow cytometry based on screening not only increased yield but also improved pullulan’s molecular weight. The adaptability of this method to other polysaccharides and its potential for genomic analysis and broader applications make it a valuable tool in bioproduction. Full article

Trilobatin (TBL) is a key sweet compound from the traditional Chinese sweet tea plant (Rubus suavissimus S. Lee). Because of its intense sweetness, superior taste profile, and minimal caloric value, it serves as an exemplary natural dihydrochalcone sweetener. It also has various health benefits, including anti-inflammatory and glucose-lowering effects. It is primarily produced through botanical extraction, which impedes its scalability and cost-effectiveness. In a novel biotechnological approach, phloretin is used as a precursor that is transformed into TBL by the glycosyltransferase enzyme ph-4′-OGT. However, this enzyme’s low catalytic efficiency and by-product formation limit the large-scale synthesis of TBL. In our study, the enzyme Mdph-4′-OGT was used to screen 17 sequences across species for TBL synthesis, of which seven exhibited catalytic activity. Notably, PT577 exhibited an unparalleled 97.3% conversion yield within 3 h. We then optimized the reaction conditions of PT577, attaining a peak TBL bioproduction of 163.3 mg/L. By employing virtual screening, we identified 25 mutation sites for PT577, thereby creating mutant strains that reduced by-products by up to 50%. This research enhances the enzymatic precision for TBL biosynthesis and offers a robust foundation for its industrial-scale production, with broader implications for the engineering and in silico analysis of glycosyltransferases. Full article

Hemp bioproducts hold great promise as valuable materials for nutraceutical and pharmaceutical applications due to their diverse bioactive compounds and potential health benefits. In line with this interest and in an attempt to valorize the Lazio Region crops, this present study investigated chemically characterized hydroalcoholic and organic extracts, obtained from the inflorescences of locally cultivated Felina 32, USO 31, Ferimon and Fedora 17 hemp varieties. In order to highlight the possible chemopreventive power of the tested samples, a bioactivity screening was performed, which included studying the antimutagenic activity, radical scavenging power, cytotoxicity in human hepatoma HepG2 cells, leakage of lactate dehydrogenase (LDH) and modulation of the oxidative stress parameters and glucose-6-phosphate dehydrogenase (G6PDH) involved in the regulation of the cell transformation and cancer proliferation. Tolerability studies in noncancerous H69 cholangiocytes were performed, too. The organic extracts showed moderate to strong antimutagenic activities and a marked cytotoxicity in the HepG2 cells, associated with an increased oxidative stress and LDH release, and to a G6PDH modulation. The hydroalcoholic extracts mainly exhibited radical scavenging properties with weak or null activities in the other assays. The extracts were usually well-tolerated in H69 cells, except for the highest concentrations which impaired cell viability, likely due to an increased oxidative stress. The obtained results suggest a possibility in the inflorescences from the Felina 32, USO 31, Ferimon and Fedora 17 hemp varieties as source of bioactive compounds endowed with genoprotective and chemopreventive properties that could be harnessed as preventive or adjuvant healing strategies. Full article

The global market for citric acid (CA) is one of the biggest and fastest expanding markets in the food industry. The CA production employing microbial bioprocessing with efficient GRAS strains and renewable waste streams is in line with the European Union binding targets for resource efficiency, sustainable consumption-production, and low-carbon technologies. In this work, the potential of three novel wild-type Yarrowia lipolytica strains (namely LMBF Y-46, LMBF Y-47 and ACA-YC 5033) regarding the production of CA and other valuable metabolites was tested on glucose-based media, and the most promising amongst the screened strains (viz. the strain ACA-YC 5033) was cultured on glucose-based media, in which part of the fermentation water had been replaced by olive-mill wastewaters (OMWs) in a novel approach of simultaneous OMW valorization and bioremediation. In the first part of this study, the mentioned strains were cultured under nitrogen-limited conditions with commercial (low-cost) glucose employed as a sole carbon source in shake-flask cultures at an initial concentration (S₀) ≈ of 50 g/L. Variable quantities of secreted citric acid (CA) and intra-cellular compounds (viz. polysaccharides and lipids) were produced. All strains did not accumulate significantly high lipid quantities (i.e., maximum lipid in dry cell weight [DCW] values ≈30% w/w were noted) but produced variable CA quantities. The most promising strain, namely ACA-YC 5033, produced CA up to c. 24 g/L, with a yield of CA produced on glucose consumed (Y_CA/S) ≈ 0.45 g/g. This strain in stirred tank bioreactor experiments, at remarkably higher S₀ concentrations (≈110 g/L) and the same initial nitrogen quantity added into the medium, produced notably higher CA quantities, up to 57 g/L (Y_CA/S ≈ 0.52 g/g). The potential of the same strain (ACA-YC 5033) to bioremediate OMWs and to produce value-added compounds, i.e., yeast cells, CA, and intra-cellular metabolites, was also assessed; under nitrogen-limited conditions in which OMWs had partially replaced tap water and significant glucose concentrations had been added (S₀ ≈ 100 g/L, simultaneous molar ratio C/N ≈ 285 g/g, initial phenolic compounds [Phen₀] adjusted to ≈1.0 g/L; these media were similar to the OMWs generated from the traditional press extraction systems) the notable CA quantity of 60.2 g/L with simultaneous Y_CA/S = 0.66 g/g, was obtained in shake flasks, together with satisfactory phenolic compounds removal (up to 19.5% w/w) and waste decolorization (up to 47.0%). Carbon-limited conditions with Phen₀ ≈ 1.0 g/L favored the production of yeast DCW (up to 25.3 g/L), with equally simultaneous interesting phenolic compounds and color removal. The fatty acid profile showed that cellular lipids were highly unsaturated with oleic, linoleic and palmitoleic acids, accounting for more than 80% w/w. This study proposed an interesting approach that could efficiently address the biotreatment of toxic effluents and further convert them into circular-oriented bioproducts. Full article

Gamma-aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the mammalian central nervous system that has a significant beneficial effect on human health. Traditional microbial GABA synthesis requires continuous oxygen supplementation. Here, a new anaerobic platform for GABA production was established with engineered C. tyrobutyricum ATCC 25755, which is considered an ideal anaerobic microbial-cell factory for bioproduction. Glutamate decarboxylase (GAD) and Class I heat-shock proteins were screened and overexpressed, generating an excellent Ct-pMAG strain for monosodium-glutamate (MSG) tolerance and GABA production, with a GABA titer of 14.26 g/L in serum bottles with the mixed substrate of glucose and MSG. Fed-batch fermentation was carried out in a 5 L bioreactor, achieving 35.57 g/L and 122.34 g/L final titers of GABA by applying the pH-free strategy and the pH-control strategy, respectively using MSG. Finally, a two-stage strategy (growth stage and bioconversion stage) was applied using glutamate acid (L-Glu) and glucose as the substrate, obtaining a 400.32 g/L final titer of GABA with a productivity of 36.39 g/L/h. Overall, this study provides an anaerobic-fermentation platform for high-level bio-GABA production. Full article