Search Results (101)

This study aimed to evaluate the production of xylose reductase (XR), an enzyme responsible for converting xylose into xylitol, by Candida tropicalis ATCC 750 using hemicellulosic hydrolysate from cashew apple bagasse (CABHM) as a low-cost carbon source. The effects of temperature, aeration, and fluid dynamics on XR biosynthesis were also investigated. The highest XR production (1.53 U mL⁻¹) was achieved at 30 °C, with 8.3 g·L⁻¹ of xylitol produced by the yeast under microaerobic conditions, demonstrating that aeration and fluid dynamics are important factors in this process. Cellular metabolism and enzyme production decreased at temperatures above 35 °C. The maximum enzymatic activity was observed at pH 7.0 and 50 °C. XR is a heterodimeric protein with a molecular mass of approximately 30 kDa. These results indicate that CABHM is a promising substrate for XR production by C. tropicalis, contributing to the development of enzymatic bioprocesses for xylitol production from lignocellulosic biomass. This study also demonstrates the potential of agro-industrial residues as sustainable feedstocks in biorefineries, aligning with the principles of a circular bioeconomy. Full article

Riboflavin (vitamin B₂) is an essential micronutrient required for all living organisms. It is naturally synthesized by plants and most microorganisms, including the bacterium Bacillus subtilis, the filamentous fungus Ashbya gossypii, and the yeast Candida famata—all of which are known to be riboflavin overproducers. The choice of production organism in industrial applications depends on factors such as yield, ease of cultivation, and the availability of genetic tools. As a result, several microorganisms are commonly used, and their relative prominence can shift over time with advances in metabolic engineering and process optimization. This review presents a comparative analysis of riboflavin biosynthesis across prokaryotic and eukaryotic systems, with a particular focus on regulatory mechanisms governing flavinogenesis. Special attention is given to recent advances in metabolic engineering strategies, including the application of CRISPR/Cas9 genome editing in Bacillus subtilis and Ashbya gossypii. In yeast systems, significant improvements in riboflavin production have been achieved primarily through the manipulation of transcriptional regulators (e.g., SEF1, SFU1, TUP1) and metabolic genes. The role of other important genes (PRS3, ADE4, ZWF1, GND1, RFE1, VMA1, etc.) in riboflavin overproduction in C. famata is described. The review also explores the use of alternative, low-cost feedstocks—including lignocellulosic hydrolysates and dairy by-products—to support more sustainable and economically viable riboflavin production. Although considerable progress has been achieved in genetic optimization and bioprocess development, further work is required to fine-tune metabolic flux and maximize riboflavin synthesis, particularly under industrial conditions. This review highlights key opportunities for future research aimed at refining metabolic interventions and expanding the use of renewable substrates for environmentally sustainable riboflavin production. Full article

The proliferation of harmful cyanobacteria, particularly Microcystis, poses significant risks to drinking and recreational water resources, especially under the influence of climate change. Conventional monitoring methods based on microscopy for harmful cyanobacteria management systems are limited in detecting toxigenic genotypes, hindering accurate risk assessment. In this study, we developed a digital droplet PCR (ddPCR)-based method for the simultaneous quantification of total and toxigenic Microcystis in freshwater environments. We targeted the secA gene, specific to the Microcystis genus, and the mcyA gene, associated with microcystin biosynthesis. Custom-designed primers and probes showed high specificity and sensitivity, enabling accurate detection without cross-reactivity. The multiplex ddPCR assay allowed for concurrent quantification of both targets in a single reaction, reducing the analysis time and cost. Application to field samples demonstrated good agreement with microscopic counts and revealed seasonal shifts in toxigenic genotype abundance. Notably, ddPCR detected Microcystis at very low densities—down to 7 cells/mL in the mixed cyanobacterial communities of field samples—even when microscopy failed, highlighting its utility for early bloom detection. This approach provides a reliable and efficient tool for monitoring Microcystis dynamics and assessing toxin production potential, offering significant advantages for the early warning and proactive management of harmful cyanobacterial blooms. Full article

Background: 5-Aminolevulinic acid (5-ALA) serves as a precursor in the heme biosynthesis pathway, resulting in the selective accumulation of protoporphyrin IX (PpIX) within glioma cells. This property facilitates fluorescence-guided resection (FGR) in high-grade gliomas (HGGs), enhancing surgical precision and oncological results. Nonetheless, its clinical implementation is restricted by factors such as accessibility, cost, and technical limitations. Methods: A systematic review of PubMed literature (2019–2024) was conducted to assess the efficacy of 5-ALA in HGG surgery compared to conventional white light microscopy. Studies focusing on non-neurosurgical applications, pediatric populations, and non-HGG indications were excluded. Results: Nineteen articles met the criteria. Recent studies indicate that 5-ALA-guided resection significantly enhances gross total resection (GTR) rates compared to white light surgery (75.4% vs. 54.3%, p < 0.001). Patients receiving 5-ALA-assisted resection exhibit enhanced progression-free survival (PFS) at 6 months (median 8.1 months compared to 5.4 months, p = 0.002) and overall survival (OS) (median 15.2 months versus 12.3 months, p = 0.008). The necessity for specialized neurosurgical microscopes equipped with blue light filters restricts accessibility, especially in low-resource environments. Recent advancements in fluorescence-enhancing technologies, particularly loupe-based systems, have demonstrated increases in fluorescence intensity by up to tenfold through direct emission. Sodium fluorescein, originally designed for ophthalmological use, has been adapted for enhancing contrast in intracranial tumors; however, its non-specific binding to serum albumin restricts its accuracy in glioma resection. Conclusions: Recent publications demonstrate that 5-ALA fluorescence-guided surgery significantly improves gross total resection rates and survival outcomes in patients with high-grade gliomas. Although it offers clinical advantages, cost and equipment constraints continue to pose substantial obstacles to broad implementation. Additional research is required to enhance fluorescence-guided techniques and increase accessibility in resource-constrained environments. Full article

Sesquiterpenols, a class of natural compounds composed of three isoprene units that form a 15-carbon skeleton with hydroxyl (-OH) group, are characterized by their volatility and potent aromatic properties. These compounds exhibit a wide range of biological activities, including antitumor, antibacterial, anti-inflammatory, anti-neurotoxic, antiviral, immunosuppressive, hepatoprotective, and cardiotonic effects. Due to their diverse physiological functionalities, sesquiterpenols serve as critical raw materials in the pharmaceutical, food, and cosmetic industries. In recent years, research on the heterologous synthesis of sesquiterpenol compounds using microbial systems has surged, attracting significant scientific interest. However, challenges such as low yields and high production costs have impeded their industrial-scale application. The rapid development of synthetic biology has introduced innovative methodologies for the microbial production of sesquiterpenol compounds. Herein, we examine the latest synthetic biology strategies and progress in microbial sesquiterpenol production, focusing on adaptive sesquiterpenol synthase screening and expression, synthesis pathway regulation, intracellular compartmentalized expression strategies, and tolerance to terpenoid-related toxicity. Critical challenges and future directions are also discussed to advance research in sesquiterpenol biosynthesis. Full article

D-tagatose is a functional sweetener with glucose-regulating and prebiotic properties, but its bioproduction from D-galactose faces many limitations, particularly the high production costs. In particular, the current biosynthesis of D-tagatose suffers from thermal instability and the substrate selectivity issues of L-arabinose isomerase (L-AI) required to convert D-galactose into D-tagatose. In this study, recombinant Escherichia coli BW25113/pQE-80L-araA_F118M/F279I expressing double mutant L-AI was immobilized to enhance its stability and reusability. The optimal conditions for whole-cell catalysis were 60 °C, pH 6.5, 5 mM Mn²⁺, and 20 h, with a yield of 55.2 g/L of D-tagatose. Immobilization with 3% sodium alginate and 2% CaCl₂ retained 90% of the production efficiency displayed by free cells. Notably, the immobilized cells exhibited enhanced heat resistance (60–70 °C) and operational stability, retaining 76% activity after five cycles. The D-tagatose production was further increased to 129.43 g/L by increasing the substrate concentration to 250 g/L. Compared to free cells, immobilized cells retained 83.6% of the initial yield up to 10 batches. This study presents a cost-effective and sustainable method for the production of D-tagatose using optimized whole-cell catalysis through immobilization, which paves the way to solve industrial challenges such as thermal instability and low substrate efficiency. Full article

Haploid breeding technology has advantages in terms of saving time and reducing labor intensity and costs. However, the low induction rate limits the application of this technology. Previous researchers found that heat shock can increase the rate of Embryo-like structures (ELSs) induction. However, molecular mechanisms underlying heat-shocked haploid induction remain poorly understood. In the current study, unfertilized ovules of watermelon were subjected to heat shock for 0–5 days and conducted transcriptomics sequencing and DIA-based proteomics sequencing. Results indicated that, in contrast to the non-heat-shock condition, the expression level of protein phosphatase 2C (PP2C), a negative regulator in abscisic acid (ABA) signal transduction pathway, was repressed, and the expression level of Sucrose-non-fermenting 1-related protein kinases (SnRK2) was activated. The activated SnRK2s are enabled to promote the accumulation of storage substances in ovules. Through analysis, the expression of many genes involved in the biosynthesis of unsaturated fatty acids and amino acids has indeed been upregulated. In conclusion, our findings demonstrate that heat shock promotes the accumulation of storage substances in unfertilized ovules by activating the signal transduction process of ABA, which correspondingly increases ELSs induction rate. Full article

Maghemite nanoparticles (NPs) were successfully developed using phenolic-rich extracts (cyanidin) from Citrus reticulata peel residues. The 11 nm maghemite NPs, obtained at 3% w/v and at 353 K, presented the optimal synthesis conditions. To improve dye adsorption performance, the synergetic adsorption behavior between these 11 nm NPs and multiwall carbon nanotubes was demonstrated. Prior to the adsorption tests, the aging effect on NPs was carefully assessed using various analytical techniques, which clearly showed the magnetite–maghemite phase transition. However, this had no impact on the cyanidin coating or adsorption properties. A remarkable percentage removal of (93 ± 3)% for methylene blue and (84 ± 3)% for methylene green was achieved in short equilibrium times of 10 and 25 min, respectively, with an optimum pH value of 5.5. Reuse experiments revealed that 90% removal for both dyes was achieved between the second to seventh regeneration cycles. Organic loading during these cycles was effectively confirmed by X-ray photoelectron spectroscopy and magnetic measurements. Dye adsorption involves a two-step mechanism: (i) electrostatic adsorption by the negative surface groups of the adsorbent (isoelectric point of 5.2) and the dye cationic groups and (ii) π–π stacking interactions between the aromatic benzene rings of the dyes, the hexagonal skeleton of the multiwall carbon nanotubes, and the phenolic ring groups of the biosynthesized sample. These results suggest that the low-cost modified phenolic adsorbent can be successfully applied to dye removal from water with promising recycling properties. Full article

Impaired function of Polymerase-γ (Pol-γ) results in impaired replication of the mitochondrial genome (mtDNA). Pathogenic mutations in the POLG gene cause dysfunctional Pol-γ and dysfunctional mitochondria and are associated with a spectrum of neurogenetic disorders referred to as POLG spectrum disorders (POLG-SDs), which are characterized by neurologic dysfunction and premature death. Pathomechanistic studies and human cell models of these diseases are scarce. SH-SY5Y cells (SHC) are an easy-to-handle and low-cost human-derived neuronal cell model commonly used in neuroscientific research. Here, we aimed to study the effect of reduced Pol-γ function using stable lentivirus-based shRNA-mediated knockdown of POLG in SHC, in both the proliferating cells and SHC-derived neurons. POLG knockdown resulted in approximately 50% reductions in POLG mRNA and protein levels in naïve SHC, mimicking the residual Pol-γ activity observed in patients with common pathogenic POLG mutations. Knockdown cells exhibited decreased mtDNA content, reduced levels of mitochondrial-encoded proteins, and altered mitochondrial morphology and distribution. Notably, while chemical induction of mtDNA depletion via ddC could be rescued by the mitochondrial biosynthesis stimulators AICAR, cilostazol and resveratrol (but not MitoQ and formoterol) in control cells, POLG-knockdown cells were resistant to mitochondrial biosynthesis-mediated induction of mtDNA increase, highlighting the specificity of the model, and pathomechanistically hinting towards inefficiency of mitochondrial stimulation without sufficient Pol-γ activity. In differentiated SHC-derived human neurons, POLG-knockdown cells showed impaired neuronal differentiation capacity, disrupted cytoskeletal organization, and abnormal perinuclear clustering of mitochondria. In sum, our model not only recapitulates key features of POLG-SDs such as impaired mtDNA content, which cannot be rescued by mitochondrial biosynthesis stimulation, but also reduced ATP production, perinuclear clustering of mitochondria and impaired neuronal differentiation. It also offers a simple, cost-effective and human (and, as such, disease-relevant) platform for investigating disease mechanisms, one with screening potential for therapeutic approaches for POLG-related mitochondrial dysfunction in human neurons. Full article

D-tagatose is an ideal sucrose substitute with potential applications in food and healthcare. The combined catalysis of polyphosphate kinase (PPK), fructose kinase (FRK), D-tagatose-6-phosphate 3-differential anisomerase (FbaA) and phytase provides a low-cost and convenient pathway for the biosynthesis of D-tagatose from D-fructose; however, there is still a problem of low catalytic efficiency that needs to be solved urgently. Therefore, this study enhanced the biosynthesis of D-tagatose by optimizing the expression levels of PPK, FRK and FbaA in a polycistronic system and knocking out the gene pfka of Escherichia coli. With 30 g/L D-fructose as a substrate, the conversion rate increased to 52%, which was the highest after 24 h. In addition, by constructing a multienzyme self-assembly system with SpyTag and SpyCatcher to improve the whole-cell catalytic ability, the conversion rate was further increased to 75%. Finally, through the fed-batch strategy, the optimal strain Ec-7 produced 68.1 g/L D-tagatose from 100 g/L D-fructose. The multienzyme cascade route reported herein provides an efficient and elegant innovative solution for the generation of D-tagatose. Full article

Background/objectives: Streptococcus agalactiae (or Group B Streptococcus, GBS) is a major cause of neonatal meningitis globally. There are 10 serotypes of GBS, which are distinguished by their capsular polysaccharide (CPS) structure, with serotypes Ia, Ib, II, III, IV and V responsible for up to 99% of infections. Currently, there are no licensed vaccines against GBS. The most developed candidates are glycoconjugate vaccines, which can be highly effective but are also expensive to produce by existing approaches and unaffordable for many parts of the world. Biosynthesis of recombinant glycans and glycoconjugates in tractable strains of bacteria offers a low-cost alternative approach to current chemical conjugation methods. Methods: In this study, we apply combinatorial hierarchical DNA assembly to the heterologous biosynthesis of GBS III, IV and V CPSs in E. coli. Each gene was removed from its native regulation, paired with synthetic regulatory elements and rebuilt from the bottom up to generate libraries of reconstituted pathways. These pathways were screened for glycan biosynthesis using serotype-specific antisera. Results: We identified several configurations that successfully biosynthesised the GBS CPSs. Furthermore, we exploited the conserved nature of the GBS CPS biosynthesis loci and the flexibility of modular DNA assembly by constructing hybrid pathways from a minimal pool of glycosyltransferase genes. We show that transferase genes with homologous function can be used interchangeably between pathways, obviating the need to clone a complete locus for each new CPS assembly. Conclusions: In conclusion, we report the first demonstration of heterologous GBS CPS IV and V biosynthesis in E. coli, a key milestone towards the development of low-cost recombinant multivalent GBS glycoconjugate vaccines. Full article

It is estimated that approximately 25% of waste remains after the apple juice pressing process. Combining this waste biomass with valuable compounds creates the potential for reuse. To create a cost-efficient ecological process without any expensive steps, the aim of this research was to examine the potential of using non-Saccharomyces yeasts (Kazachstania barnettii D1, Hanseniaspora uvarum D9, and Wickerhamomyces anomalus D11) for the low-temperature valorisation of apple pomace. The scope encompassed characteristics of apple pomace and the evaluation of yeast growth and metabolic activity, including carbohydrate consumption, enzymatic activity, and the biosynthesis of volatile organic compounds. Moreover, the effect of inoculum size on biomass increases and the productivity of metabolites during the fermentation of apple pomace were evaluated. To investigate the potential intensification of the process, the experiment was performed on hydrolysed and untreated apple pomace. The obtained results indicate that yeast growth was satisfactory regardless of the inoculum size in both fermentation media. Various activities of peptidases, esterases, phosphatases, and glucosidases were observed. The yeast isolates presented metabolic activity during the process which was confirmed by the production of ethanol and acetic acid. Moreover, a significant amount of volatile organic compounds, especially esters, were synthesised, which have a positive impact on the sensory profile of fermented apple pomace. In general, the hydrolysis of apple pomace did not result in better yeast activity and the formation of aroma compounds. Full article

Chaetoglobosin A (CheA), a typical structure of the cytochalasin family, exhibits outstanding efficacy against a variety of tumor cells and plant pathogens. However, its low yield and high production cost are major obstacles limiting its wide application. In order to increase CheA yield, an engineered strain was established by overexpressing CgMfs, the gene encoding the MFS family’s efflux pump, on chassis cells lacking CgXpp1, which have been shown to act as a negative regulator of CheA biosynthesis. As expected, the engineered strain significantly boosted CheA production from 63.19 to 265.93 mg/L after incubation in PDA medium for 10 d, whereas the yield of the engineered strain was remarkably enhanced 2.93-fold compared with the wild type, following 10 d of cultivation utilizing potato starch industrial waste. The addition of metal ions had a positive effect on CheA production, with Cu²⁺ being the most effective and improving production to 176.92 mg/L. The optimal fermentation conditions were determined by response surface optimization, and under the optimal conditions, the engineered strain could stably produce CheA with a yield of 197.58 mg/L. This study provided the conditions for reducing production costs while increasing CheA production, as well as new strategies and insights for the production of the target compound. Full article

The rising demand for sustainable materials has led to a significant focus on developing resources from renewable systems, particularly through the integration of biological processes. Bacterial cellulose (BC) has emerged as a highly promising biomaterial, gaining attention across multiple industries, such as food, pharmaceuticals, materials science, and textiles, due to its renewable, biodegradable, and eco-friendly characteristics. Within the fashion industry, bacterial cellulose (BC) biofabrication presents a groundbreaking method for producing sustainable textiles and vegan leather. This systematic review emphasizes BC’s pivotal role in advancing sustainable materials, addressing challenges like low yields, strain instability, and high production costs, and exploring innovative biofabrication techniques to overcome these barriers. Current advancements aim to enhance the thickness, uniformity, and mechanical properties of BC layers by optimizing the environmental and nutritional conditions during Komagataeibacter cultivation and leveraging coculturing methods. Furthermore, recent innovations in synthetic biology and genetic engineering have opened new avenues for improving BC biosynthesis, making it a viable solution for the sustainable fashion industry. This review explores three core topics: (1) bacterial cellulose and its applications, (2) the biofabrication of BC for vegan leather, and (3) emerging innovations and patents utilizing bacterial cellulose as a sustainable industrial biomaterial. Full article

Fumonisins, a class of mycotoxins predominantly produced by Fusarium species, represent a major threat to food safety and public health due to their widespread occurrence in staple crops including peanuts, wine, rice, sorghum, and mainly in maize and maize-based food and feed products. Although fumonisins occur in different groups, the fumonisin B series, particularly fumonisin B1 (FB1) and fumonisin B2 (FB2), are the most prevalent and toxic in this group of mycotoxins and are of public health significance due to the many debilitating human and animal diseases and mycotoxicosis they cause and their classification as by the International Agency for Research on Cancer (IARC) as a class 2B carcinogen (probable human carcinogen). This has made them one of the most regulated mycotoxins, with stringent regulatory limits on their levels in food and feeds destined for human and animal consumption, especially maize and maize-based products. Numerous countries have regulations on levels of fumonisins in foods and feeds that are intended to protect human and animal health. However, there are still gaps in knowledge, especially with regards to the molecular mechanisms underlying fumonisin-induced toxicity and their full impact on human health. Detection of fumonisins has been advanced through various methods, with immunological approaches such as Enzyme-Linked Immuno-Sorbent Assay (ELISA) and lateral flow immunoassays being widely used for their simplicity and adaptability. However, these methods face challenges such as cross-reactivity and matrix interference, necessitating the need for continued development of more sensitive and specific detection techniques. Chromatographic methods, including HPLC-FLD, are also employed in fumonisin analysis but require meticulous sample preparation and derivitization due to the low UV absorbance of fumonisins. This review provides a comprehensive overview of the fumonisin family, focusing on their biosynthesis, occurrence, toxicological effects, and levels of contamination found in foods and the factors affecting their presence. It also critically evaluates the current methods for fumonisin detection and quantification, including chromatographic techniques and immunological approaches such as ELISA and lateral flow immunoassays, highlighting the challenges associated with fumonisin detection in complex food matrices and emphasizing the need for more sensitive, rapid, and cost-effective detection methods. Full article