The growing consumer demand for functional fruit juices has emphasized the need for targeted enzyme applications in juice processing. Among these, xylanase has emerged as a critical enzyme for improving the extraction and quality of juices from xylan-rich fruits. However, despite its potential, xylanase remains insufficiently characterized for functional juice production, and further evaluation is necessary. In the present study, a screening assay identified Aspergillus tubingensis FAT35 as the most efficient xylanase-producing isolate among a set of fungal strains. Solid-state fermentation using agro-industrial residues was optimized, resulting in high-yield enzyme production: 550 U mL-1, which corresponds to 2333 U g-1. The enzyme exhibited high xylanase activity with negligible pectinase and cellulase side activities. Application of the xylanase to orange and pineapple juices significantly improved juice yield (17% and 3%), sugar content (22% and 39%) and clarity (2% and 22%). Furthermore, treated juices showed enhanced antioxidant and antimicrobial activities. These functional improvements were attributed to the formation of the prebiotic oligosaccharide xylotetraose (X4), indicating added health value. The optimized xylanase from A. tubingensis FAT35 demonstrates strong potential for use in the fruit juice industry. Its ability to enhance both technological parameters and bioactive properties of juices highlights its role in the development of functional beverages and underscores the importance of further testing and application of xylanases in food biotechnology. © 2025 Society of Chemical Industry.

Brown seaweeds are marine bioresources rich in bioactive compounds such as carbohydrates, proteins, pigments, fatty acids, polyphenols, vitamins, and minerals. Among these substances, brown algae-derived polysaccharides (alginate, fucoidan, and laminarin) have promising industrial prospects owing to their distinctive structural features and diverse biological activities. Consequently, processing technologies have advanced substantially to address industrial requirements for biopolymer quality, cost-effectiveness, and sustainability. Over the years, significant progress has been made in developing various advanced methods for the sake of extracting, purifying, and structurally characterizing polysaccharides. Aside from that, numerous studies reported their broad spectrum of biological activities, such as antioxidant, anti-inflammatory, anticoagulant, and antimicrobial properties. Furthermore, these substances have various industrial, pharmaceutical, bioenergy, food, and other biotechnology applications. The present review systematically outlines the brown algae-derived polysaccharides treatment process, covering the entire value chain from seaweed harvesting to advanced extraction methods, while highlighting their biological activities and industrial potential as well.

The context of food science and biotechnology is undergoing a profound transformation, characterized by an evolutionary shift from conventional large-scale fermentation to precision biomanufacturing, positioning Lactic Acid Bacteria (LAB) as versatile cellular biofactories for next-generation functional foods. This review analyzes the evolutionary role of LAB, their utilization as probiotics, and the technological advances driving this shift. This work also recognizes the fundamental contributions of pioneering women in the field of biotechnology. The primary methodology relies on the seamless integration of synthetic biology (CRISPR-Cas editing), Multi-Omics analysis, and advanced Artificial Intelligence/Machine Learning, enabling the precise, rational design of LAB strains. This approach has yielded significant findings, including successful metabolic flux engineering to optimize the biosynthesis of high-value nutraceuticals such as Nicotinamide Mononucleotide and N-acetylglucosamine, and the development of Live Biotherapeutic Products using native CRISPR systems for the expression of human therapeutic peptides (e.g., Glucagon-like Peptide-1 for diabetes). From an industrial perspective, this convergence enhances strain robustness and supports the digitalized circular bioeconomy through the valorization of agri-food by-products. In conclusion, LAB continue to consolidate their position as central agents for the development of next-generation functional foods.

ABSTRACTChitin, the natural biopolymer of the world next to cellulose, is a modified biodegradable polysaccharide. Chitosan, as the major derivative of chitin, is the product of N‐deacetylated chitin. Chitosan is an important biopolymer in nature and the only positively charged (cationic) polysaccharide. Chitosan has many utilizations in sustainable agriculture and food systems, in particular, improving plant resistance to environmental stresses like water deficit, salt, high temperature, cold, heavy metal, etc., as well as biotic stresses such as pest and plant pathogens. In addition, this natural biopolymer is used in different industries such as paper, food (processing, packaging, and preservation), pharmaceuticals, biodiesel, and other uses like wastewater treatment and environmental protection. Chitosan gained significant interest for its safety, antifungal, antibacterial, biodegradability, biocompatibility, and antioxidant activities due to its rich amino and hydroxyl groups. The commercial value of chitosan is due to the valuable properties of its soluble derivatives, which are suitable in food processing, cosmetics, nano and biotechnology, environmental, and textile production. In this review, we will consider the effectiveness of chitosan in the performance of agriculture, herbal products, nutraceuticals, and food systems, like improving biologically active compounds in herbal plants as elicitor; the characteristics of chitosan and chitosan‐based biopolymers have been mentioned.

ABSTRACTLacticaseibacillus rhamnosus GG (LGG) is one of the most extensively studied probiotic strains, widely used in food and health applications. However, the absence of efficient, precise genome editing methods has limited its broader potential and functional versatility. Here, we present an endogenous type II‐A CRISPR‐Cas genome editing workflow for LGG designed for functional strain construction. Using a plasmid interference assay together with single‐nucleotide substitutions, we confirm the precise PAM requirement as 5′‐NGAAA‐3′. We pair a synthetic sgRNA cassette with homology‐directed repair donors to enable targeted deletions and insertions across multiple loci, achieving modest but practically relevant editing efficiencies (11.1‐25.0% of recovered transformants) that support routine strain construction. Using this optimised and precise genome engineering method, we generated a β‐glucuronidase (GUS)‐expressing LGG strain for robust strain tracking within complex microbial communities. This work removes barriers to LGG engineering, expands the probiotic CRISPR toolkit, and provides broadly applicable strategies for designing next‐generation probiotics with applications in food biotechnology and microbial therapeutics.

Fungal spoilage remains a major challenge in the food industry, driven by the high adaptive capacity of molds and their ability to colonize diverse food matrices. Culture collections play a key role in documenting this diversity and providing access to well-characterized strains for research and industrial applications. This study presents a curated set of 60 mold isolates obtained from spoiled food products in Poland between 2019 and 2024 and subsequently deposited in the Collection of Industrial Microorganisms (KKP)—Microbiological Resources Center, Department of Microbiology, Prof. Wacław Dąbrowski Institute of Agricultural and Food Biotechnology—State Research Institute. The isolates originated from fruit-based products, cereal- and flour-derived items, dairy products and meat, reflecting the wide range of substrates susceptible to fungal contamination. Taxonomic identification based on ITS sequencing revealed representatives of common food spoilage genera, including Penicillium, Aspergillus, Mucor, Alternaria, Cladosporium and others, together with less frequently reported taxa and physiologically resilient species such as Xeromyces bisporus and Paecilomyces niveus. The dataset highlights the occurrence of xerophilic, psychrotolerant and otherwise stress-resistant molds capable of persisting under reduced water activity, low temperature or modified-atmosphere conditions. By documenting the diversity and origins of these isolates, this work expands the reference resources available for studies on fungal ecology, spoilage mechanisms and contamination pathways in food environments. The strains preserved in the KKP collection constitute a valuable foundation for future research aimed at improving food safety, developing targeted detection methods and assessing antifungal strategies.

Fumonisins, a major class of mycotoxins, pose significant health risks to humans and animals due to their widespread contamination and potent toxicity. Recent advances in molecular biology, biochemistry, and enzymology have greatly enhanced the understanding of fumonisin biosynthesis and its genetic regulation. The key biosynthetic genes are typically organized in clusters and regulated by specific transcription factors; increasing evidence also highlights the involvement of complex transcriptional and epigenetic mechanisms. Environmental factors such as nitrogen, carbon, and pH also modulate these regulatory networks. Despite substantial progress, critical gaps remain in fully elucidating the regulatory pathways that control fumonisin production. This review synthesizes current knowledge regarding fumonisin biosynthesis, gene clusters, and multi-level regulatory mechanisms, while emphasizing recent trends, existing challenges, and potential applications in food safety and biotechnology to enhance food security and promote sustainable development.

Aspergillusflavus is a globally important human pathogen and agricultural contaminant, while its domesticated relative A.oryzae is widely used in food fermentation and biotechnology. Despite their importance, the evolutionary relationship, population structure and domestication history of these fungi remain unresolved. Here, we present the first global population genomic analysis of 639 A.flavus and A.oryzae isolates from clinical, environmental and food-fermentation sources across multiple continents. Our analyses reveal a complex evolutionary landscape comprising well-separated clades interspersed with highly admixed mosaic groups and potential evidence for multiple independent domestication events giving rise to A.oryzae. Clinical A.flavus isolates are distributed across several clades and mosaic groups, some overlapping with fermentation strains, highlighting an apparent role of domestication and admixture in shaping pathogen diversity. These results challenge current species boundaries and provide a framework for understanding evolutionary history, taxonomy and pangenomic architecture in these fungi, with broad implications for pathogenicity, food safety, biocontrol and metagenomic surveillance.

Monosaccharide profiling and osmoprotective effects of camel milk: Implications for protein stability and therapeutic use.

Objectiveα-Amylases represent a class of industrially critical enzymes widely employed in food processing, animal feed, and biotechnology sectors. While microbial sources, particularly Bacillus species, serve as preferred production platforms due to their efficiency and scalability, there remains a pressing need to discover novel strains with enhanced enzymatic properties from underexplored environments. This study investigated the isolation and characterization of high-performance α-amylase-producing bacteria from diverse ecological niches across Iran, including Golestan, Mazandaran, Gilan, Kurdistan, and Isfahan, with particular emphasis on their potential application in animal feed industries.MethodsStrains capable of producing amylase were extracted from environmental samples in Iran, subjected to screening through starch hydrolysis, and subsequently optimized for their enzymatic activity. The chosen strains underwent morphological and genetic characterization, including 16S/rpoB and whole-genome sequencing, followed by the purification of the enzyme and in silico analysis.ResultsThrough systematic screening of sixty bacterial isolates obtained from soil, water, and industrial effluent samples, four promising strains were selected based on their superior productivity indices (PI > 1.8) and enzymatic activity profiles. Among these, strains S1 and S3 (isolated from the Kuhrang water source and Gavkhouni Wetland in Isfahan province, respectively) demonstrated exceptional α-amylase production capabilities (reaching 34,121 U/g under optimal conditions) along with remarkable stability across broad pH (4−9) and temperature (30−80°C) ranges. Comprehensive genomic characterization, including whole-genome sequencing and phylogenetic analysis, identified these isolates as novel variants of Bacillus spizizenii, marking the first report of this species’ α-amylase potential from Iranian ecosystems, specifically originating from Isfahan province. Further analysis revealed a 1980 bp GH13-family α-amylase gene encoding a 62 kDa enzyme with 93.6% sequence similarity to AmyE of B. subtilis, while molecular docking studies demonstrated strong binding affinities (−8.4 kcal/mol) with maltotetraose, supported by critical interactions with catalytic residues.ConclusionsThe combination of robust enzymatic activity, exceptional environmental tolerance, and confirmed genetic basis positions these B. spizizenii strains as highly promising candidates for industrial enzyme applications. This work not only expands the known diversity of high-performance α-amylase producers but also provides valuable insights into the biotechnological potential of underutilized Bacillus species from unique geographical sources.

Genetically Modified Foods (GMFs) have become one of the most controversial innovations in food production and biotechnology. Public concerns regarding the safety, ethical considerations, and health impacts of GMFs have fueled widespread debate. This study explores the impact of social media exposure on individuals’ knowledge of and attitudes toward Genetically Modified Foods and how these factors influence their purchase intention. The findings from an online survey of 467 participants in Saudi Arabia show that higher levels of social media exposure are associated with increased knowledge and stronger perceptions of both benefits and risks of GMFs. Purchase intentions, however, are driven primarily by perceived benefits (positively) and perceived risks (negatively), while knowledge exerts an indirect effect through these attitudinal components.

Ecdysteroids are polyhydroxylated sterols initially identified in insects but later reported in numerous terrestrial plants, including Ajuga, Rhaponticum, and Cyanotis. These metabolites display diverse biological properties such as anabolic, adaptogenic, antioxidant, antimicrobial, and antitumor effects, making them highly relevant for pharmaceutical and nutraceutical applications. However, their occurrence in aquatic plants remains poorly studied. This review highlights the potential of fast-growing macrophytes, particularly Lemna minor (duckweed), as promising sources of ecdysteroids and other bioactive compounds including flavonoids and amino acids. Available literature is critically analyzed to compare existing findings and to assess their implications for food technology, pharmacy, and biotechnology. By integrating current evidence, this article positions Lemna minor as a model system for exploring aquatic plant-based metabolites. The review also outlines future research priorities and industrial opportunities for sustainable production of nutraceutical and pharmaceutical agents from aquatic sources.

The rapidly developing mass-market of functional foods needs new delivery methods for biologically active substances. This article describes an innovative dietary biopolymer-based food capsule. The methodology involved modern food biotechnology methods, analytical chemistry, and computer modelling. The encapsulation procedure started with preparing the bioactive components followed by gel matrix formation and stabilization. The biopolymer system demonstrated good prospects for encapsulating alkaloids, e.g., berberine, in an intestinal simulation in vitro. The biopolymer gel proved resistant to acidic environments and provided a prolonged release of the active substance, releasing over 80% of the encapsulated berberine. This effective technology can be recommended for commercial production of functional foods.

Leuconostoc mesenteroides is a key microorganism in food biotechnology, valued for its production of flavor-forming metabolites and exopolysaccharides, and its inclusion in starter cultures and biocatalytic systems. However, the application of advanced genetic tools to L. mesenteroides remains hindered by multiple barriers, including inefficient DNA transfer, elevated endogenous nuclease activity, and restriction–modification systems sensitive to plasmid methylation patterns. As a result, even widely accepted electroporation methodologies often yield inconsistent or irreproducible transformation results, limiting the strain’s amenability to metabolic engineering and synthetic biology applications. In this study, a reproducible electroporation protocol for the L. mesenteroides strain H32-02 Ksu is developed and experimentally validated. The protocol concept relies on the sequential optimization of key process steps: targeted weakening of the cell wall followed by osmotic protection, the development of a gentle electrical stimulus that ensures membrane permeability without critical damage, and the creation of recovery conditions that minimize loss of viability and degradation of incoming DNA. Matching plasmid methylation to the recipient’s restriction profile proved critical: choosing a source for plasmid DNA production with a compatible methylation pattern dramatically increased the likelihood of successful transformation. In our case, the selection of an E. coli strain with a more suitable methylation profile increased the yield of transformants by 3.5 times. It was also shown that reducing the pulse voltage increase transformant number by 3 times. The combined optimization resulted in an approximately 40-fold increase in transformation efficiency compared to the baseline level and, for the first time, provided consistently reproducible access to transformants of this strain. The highest transformation efficiency was achieved: 8 × 102 CFU µg−1 DNA. The presented approach highlights the strain-specificity of barriers in Leuconostoc and forms a technological basis for constructing strains with desired properties, expressing heterologous enzymes, and subsequently scaling up bioprocesses in food and related industries. The methodological principles embodied in the protocol are potentially transferable to other lactic acid bacteria with similar limitations.

Microbial resources are essential for food biotechnology, where they contribute to product quality, process efficiency, and safety. Their long-term preservation is critical for culture collections (CCs) and microbial biological resource centers (mBRCs). This study investigates the effects of freeze-drying on five lactic acid bacteria (LAB) strains from the Unimore Microbial Culture Collection (UMCC), comparing non-lyophilized and lyophilized cultures under two pre-freezing protocols: overnight incubation at -20 °C (PF-20) and 2-hour incubation at -80 °C (PF-80). Viability, fermentative performance, metabolite production, and cell integrity were assessed using microbial counts, HPLC profiling, and SEM imaging, respectively. Lactiplantibacillus plantarum UMCC 2996 maintained over 95% viability and stable fermentative traits across all conditions. In contrast, Fructilactobacillus sanfranciscensis UMCC 2990 and Leuconostoc citreum UMCC 3011 showed significant viability losses (down to 54.45% and 73.89%, respectively) and altered metabolic profiles, particularly under PF-80. SEM analysis confirmed structural damage in sensitive strains, with visible cellular debris and membrane wrinkling. This study provides novel perspectives on the customized assessment of freeze-drying protocols for sourdough-derived LAB strains with industrial potential, confirming the need for tailored preservation strategies to ensure the long-term functionality of LAB strains in CCs and mBRCs.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00284-025-04673-5.

Antimicrobial resistance (AMR) remains a critical global challenge, requiring novel complementary strategies beyond antibiotic development. Probiotic-fermented foods (PFFs) offer an emerging, low-cost approach to mitigate AMR risk through ecological, molecular, and immunological mechanisms. This review integrates mechanistic insights, clinical evidence, and translational frameworks linking PFFs to antimicrobial stewardship. Key mechanisms include colonization resistance, nutrient and adhesion-site competition, production of antimicrobial metabolites, such as bacteriocins, hydrogen peroxide, and organic acids and Quorum-quenching-sensing activities that suppress pathogen virulence. Randomized clinical trials indicate that fermented diets and probiotic supplementation can improve microbiome diversity, reduce inflammatory cytokines, and decrease antibiotic-associated diarrhea, though direct AMR outcomes remain underexplored. Evidence from kefir, kombucha, and other microbial consortia suggests potential for in vivo pathogen suppression and reduced infection duration. However, safe translation requires standardized starter-culture genomics, resistome monitoring, and regulatory oversight under QPS/GRAS frameworks. Integrating PFF research with One Health surveillance systems, such as the WHO GLASS platform, will enable tracking of antimicrobial consumption and resistance outcomes. Collectively, these findings position PFFs as promising adjuncts for AMR mitigation, linking sustainable food biotechnology with microbiome-based health and global stewardship policies.

Lipase is a biocatalyst capable of catalyzing the hydrolysis or synthesis of triglycerides. It is widely distributed in animals, plants, and microorganisms. Owing to its high efficiency, specificity, and mild reaction conditions, lipase has demonstrated significant value in food processing, nutritional enhancement, and waste treatment. This paper reviews the sources of lipase and its applications in food processing, flavor improvement, and nutritional enhancement, as well as discusses its future development trends. Lipase can be utilized in oil modification, dairy product processing, baked goods, meat product processing, and other areas to enhance food quality and nutritional value. Furthermore, lipase exhibits substantial potential in the utilization of food waste resources and the development of functional foods. With advancements in enzyme engineering technology and deeper insights into the mechanism of lipase, its application in food biology will become increasingly extensive and profound.

In order to define the term GMO, different scientific definitions and legal explanations are available. In theregulation process of GM foods, the US and EU legal frameworks are based on the methodologiesthemselves. Currently, for the production of GMOs, several genome editing tools are available. Along withdifferent site-directed nucleases (ZFN, TALENs, etc.), RNAi and CRISPR/Cas9 have proven to be the veryeffective tools for genome editing. According to the current EU legislative, introduced in 2018,CRISPR/Cas9 and RNAi techniques are regulated as methods that produce GMOs, because themethodology of the process itself resembles the traditional breeding methods. In the past few years, a largenumber of scientific publications have confirmed that CRISPR/Cas9 and RNAi technology produce GMOs,supporting and suggesting that the legislation policies in the EU and especially in the USA have to beelaborated. Besides, a huge public pressure makes it difficult to develop and implement new methodologiesfor GMO production. For this reason, ELSI society is responsible to investigate and question whether thenew genetic engineering techniques produce GMO food that is safe for human consumption.

Dissecting native plasmid functions in Lactococcus lactis: Synergistic regulation of metabolism and industrially relevant traits.

Biopolymer composites with halloysite nanotubes for multifunctional enhancement in food packaging: A review.