Strain development, bioprocess optimization, and food-structure design for sustainable mycoprotein production from edible filamentous fungi.

From Bioreactor to Market: Opportunities and Challenges of Animal-Free Proteins from Precision Fermentation, Cell Culture and Molecular Engineering

This work explores the development of animal-free proteins produced in laboratory settings using innovative technologies such as precision fermentation, submerged fermentation, plant cell culture, and molecular engineering, which enable the production of high-quality proteins without relying on animal farming or large-scale traditional agriculture.

Microbial Production of Alternative Proteins for Food Applications: Advances, Industrial Challenges, and Regulatory Status.

The escalating global population and the environmentally inefficient nature of livestock-based protein production are intensifying demand for sustainable and scalable protein alternatives. Microbial biosynthesis, employing engineered cell factories, represents a pivotal strategy for producing functional proteins with a reduced ecological footprint. This review comprehensively examines the biosynthesis of alternative proteins (APs) via microbial precision fermentation, encompassing diverse categories including coloring proteins, flavoring and taste proteins, structuring and texturizing proteins, nutritional and functional proteins, food processing and enabling proteins, and special functional proteins. Enabling technologies, from fermentation feedstock and microbial host selection to genome/metabolic engineering, bioprocess optimization via response surface methodology/artificial neural networks, and downstream purification, are critically analyzed. Emerging strategies demonstrate substantial progress in enhancing microbial titers, achieving functional mimicry, and advancing regulatory readiness. However, persistent challenges include precise flavor replication, nutritional completeness, and food safety concerns such as allergenicity and process contaminants. Potential solutions, including advanced metabolic engineering, refined protein extraction, biocontainment strategies, and transparent regulatory frameworks, are discussed. By integrating technological innovation with targeted application mapping and regulatory foresight, this review outlines a roadmap toward scalable, safe, and functionally robust microbial AP platforms, thereby contributing to the transition toward a sustainable food system.

Bioprocess Engineering for Enzyme Manufacturing: A Systems Approach to Production, Purification, and Formulation

Enzymes constitute one of the largest and fastest-growing sectors of industrial biotechnology, serving as environmentally sustainable biocatalysts in food and beverage processing, fermentation industries, biofuel production, detergents, textiles, leather processing, pulp and paper manufacturing, pharmaceuticals, nutraceuticals, animal feed, and environmental remediation. Their exceptional catalytic efficiency, substrate specificity, and ability to operate under mild reaction conditions have enabled the replacement of numerous conventional chemical catalysts, thereby reducing energy consumption, hazardous waste generation, and overall environmental impact. Continuous advances in microbial biotechnology, metabolic engineering, protein engineering, synthetic biology, and precision fermentation have significantly improved enzyme productivity, catalytic performance, process economics, and industrial scalability. This review comprehensively examines four interconnected pillars of industrial enzyme biotechnology: (i) microbial fermentation technologies, (ii) downstream processing and industrial scale-up, (iii) bioprocess optimization, and (iv) enzyme formulation and delivery systems. Current fermentation strategies—including submerged fermentation (SmF), solid-state fermentation (SSF), batch, fed-batch, and continuous cultivation—are critically evaluated with respect to productivity, substrate utilization, scalability, and commercial feasibility. Recent advances in downstream processing, encompassing biomass separation, enzyme concentration, purification, stabilization, drying, and quality assurance, are discussed with emphasis on maximizing product recovery while minimizing activity loss and manufacturing costs. Modern bioprocess optimization approaches—including Design of Experiments (DoE), Response Surface Methodology (RSM), systems biology, metabolic engineering, artificial intelligence-assisted process control, digital bioprocessing, and Process Analytical Technology (PAT)—are highlighted for their roles in improving enzyme yield, productivity, robustness, and manufacturing efficiency. Furthermore, recent developments in enzyme formulation technologies, including liquid concentrates, powder formulations, immobilized enzymes, nanoformulations, encapsulation, and controlled-release delivery systems, are reviewed for their ability to enhance enzyme stability, shelf life, transportability, and industrial performance. By integrating upstream production, downstream recovery, process optimization, and formulation technologies into a unified framework, this review provides a holistic perspective on industrial enzyme manufacturing. The article serves as a comprehensive resource for researchers, industrial microbiologists, biochemical engineers, and biotechnology professionals seeking to develop economically viable, sustainable, and high-performance enzyme production systems. Future progress is expected to arise from the convergence of synthetic biology, machine learning, precision fermentation, digital bioprocessing, and circular bioeconomy concepts, thereby enabling next-generation sustainable enzyme manufacturing.

Fungal consortium–driven valorization of agro-waste for sustainable co-production of industrial enzymes through integrated statistical bioprocess modelling

Precision fermentation and recombinant proteins as enabling technologies for scalable cellular agriculture.

Cellular agriculture has emerged as a promising strategy for producing animal-derived food components through controlled biological processes while reducing the environmental and ethical burdens associated with conventional livestock production. Among its enabling technologies, precision fermentation and cultivated-cell systems offer complementary advantages but continue to face challenges related to production costs, scalability, and functional performance. Increasingly, hybrid cellular agriculture approaches combining precision-fermented proteins, cultivated cells, and plant-derived matrices are being explored to overcome these limitations and accelerate commercialization. This review examines recombinant proteins as critical enabling components within these integrated systems. Advances in microbial expression platforms, including prokaryotic hosts such as Escherichia coli and Bacillus subtilis and eukaryotic hosts such as Saccharomyces cerevisiae and Komagataella phaffii, are critically evaluated regarding protein yield, product quality, regulatory suitability, downstream processing, and techno-economic feasibility. Industrial-scale fermentation capacities up to 80,000 L demonstrate the growing potential for large-scale recombinant protein production. Applications of recombinant proteins in edible scaffolds, serum-free culture media, extracellular matrix alternatives, and functional food ingredients are discussed alongside their associated technical and regulatory challenges. Ultimately, recombinant proteins are identified as integrative elements bridging acellular and cell-based production systems, supporting the development of scalable, economically viable, and sustainable cellular agriculture.

Research on the Production of Single-Cell Protein from Gibberellin Fermentation Waste Liquid Using Microbial Methods

Efficient monitoring and intelligent regulation of industrial bioprocesses are increasingly important for sustainable resource utilization and advanced manufacturing systems. This study investigates the production of single-cell protein (SCP) from gibberellin fermentation waste liquid using the unconventional yeast Nectaromyces rattus, aiming to achieve simultaneous pollutant removal and biomass valorization. Bottle culture experiments and tank reactor simulations were conducted to evaluate the effects of wastewater dilution ratio and glucose supplementation on microbial growth, nitrogen assimilation, and SCP production. The results indicate that under a 12-fold wastewater dilution with 10 g/L glucose addition, corresponding to an optimized C/N ratio of approximately 18:1, the dry cell weight reaches 192.336 g/L and the ammonia nitrogen removal efficiency attains 85.54%. Furthermore, systematic analyses of strain adaptation, fermentation optimization, and quality evaluation demonstrate the feasibility of integrating biological treatment with high-value protein production for industrial wastewater recycling. The proposed strategy establishes an effective framework for sustainable biomanufacturing and process intensification. In addition, its reliance on real-time reactor monitoring, multiparameter process control, and intelligent optimization provides valuable engineering references for electromagnetic sensing technologies, wireless monitoring architectures, antennaassisted industrial instrumentation, and advanced signal acquisition systems used in next-generation bioprocess automation.