Ganoderma lucidum is a medicinal mushroom widely recognized as a source of biomolecules with pharmacological properties, however, little is known about the factors that influence the synthesis of bioactive proteins by this fungus when cultivated under submerged fermentation. The objective of this work was to evaluate the production of mycelial biomass and intracellular proteases and protease inhibitors by G. lucidum cultivated under different submerged fermentation conditions. The cultivation was carried out in a medium composed of glucose (10 or 20 g.L-1), soy peptone (2.5 or 5 g.L-1) and yeast extract (5 g.L-1), with incubation under agitation (120 rpm) and non-agitation, totaling 8 experimental conditions. Biomass production was determined from the dry weight, while glucose consumption was estimated by quantification of reducing sugars. The proteins were extracted in NaCl (0.15 M), and the protein extracts were submitted to protein quantification by the Bradford method, total proteolytic activity using azocasein, caseinolytic and fibrinolytic activity in Petri dishes, activity of serine (trypsin and chymotrypsin) and cysteine (papain) protease inhibitors. Cultivation in agitated condition showed higher biomass production with a maximum value of 7 g.L-1, in addition to higher activities of trypsin, chymotrypsin and papain inhibitors, with 154 IU.mg-1, 153 IU.mg-1 e 343 IU.mg-1 of protein, respectively. The non-agitated condition showed a greater potential for obtaining proteins, total proteases, caseinolytic and fibrinolytic enzymes, with maximum values of 433 mg.g-1 of extract, 71 U.mL-1 of extract, 63.62 mm2 and 50.27 mm2, respectively. Thus, a medium composed of soy peptone, yest extract and glucose in a 1:2:4 proportion is recommended, under agitation to produce protease inhibitors, and the non-agitated condition when the target is, mainly caseinolytic and fibrinolytic enzymes.

The industrial transition to advanced biofuels is currently limited by the metabolic constraints and low inhibitor tolerance of wild-type microbial hosts. This review justifies the necessity of Precision Fermentation (PF) as the pivotal technological framework to overcome these barriers, providing a systematic synthesis of high-resolution genetic tools and intelligent bioprocess architectures. We analyze how the integration of CRISPR-Cas9, retron-mediated recombineering, and synthetic regulatory circuits enables the development of specialized microbial “chassis” capable of achieving 10- to 100-fold higher yields compared to native organisms, with industrial titers reaching 50 g/L for isobutanol and 25 g/L for farnesene. A major novelty of this work is the critical evaluation of Artificial Intelligence (AI), Soft Sensing, and Digital Twins in orchestrating real-time metabolic control and mitigating the toxic effects of advanced alcohols and drop-in hydrocarbons (C15–C20). Furthermore, the study concludes that the “scale-out” modular strategy, when integrated into hybrid thermochemical-biochemical biorefineries, allows for the full valorization of C5/C6 sugars and lignin, achieving a Minimum Selling Price (MSP) competitive with fossil fuels. By mapping the synergy between advanced metabolic engineering and data-driven process optimization, this review establishes PF as an indispensable driver for achieving carbon-neutral and carbon-negative energy systems in the circular bioeconomy.

The global functional food market continues to expand, and edible mushrooms are emerging as high-value ingredients due to their rich nutritional profile, particularly their high protein content, balanced amino acid composition, and dietary fiber. This growing industrial interest is reflected in the registration of more than 322 patents in the past five years according to the Derwent Innovation patent database. Recent advances include the integration of precision mycology (PM) and omics-based approaches, such as CRISPR-Cas9, into solid-state fermentation and submerged fermentation, enabling improvements in natural umami flavor and bioactive composition. Innovative products, including meat analogues with fibrous textures, functional beverages such as kombucha and juices, and fermented dairy products such as yogurts and cheeses, have been formulated to deliver prebiotic, antioxidant, and immunomodulatory properties. Future trends indicate a shift towards the production of high-value nutraceutical peptides and biomass, together with the adoption of artificial intelligence (AI) and the Internet of Things (IoT) to enhance bioreactor automation and scalability. Nevertheless, significant challenges remain, including regulatory constraints, the scarcity of clinical validation in humans, and the need for strict control over the bioaccumulation of heavy metals in mushroom-derived raw materials. Addressing these gaps will be critical for advancing regulatory frameworks, improving industrial standardization, and supporting the translational development of mushroom-based functional foods.

Background: Actinobacteria are major producers of antibacterial and antifungal metabolites and are growing their search for substances of biotechnological interest, especially for use in agriculture, among other applications. The Amazon is potentially rich in actinobacteria; however, almost no research studies exist. Thus, we present a study of the occurrence and antifungal potential of actinobacteria from the rhizosphere of Inga edulis, a native South American plant and one that is economically useful in the whole of the Amazon. Methods: Among the 64 actinobacteria strains isolated from the rhizosphere of three Inga edulis plants, 20 strains were selected and submitted to dual-culture assays against five important phytopathogenic fungi and morphological and 16S rRNA gene analyses. Two strains, LaBMicrA B270 and B280, were also studied for production curves of metabolic extracts and antifungal activities, including their minimum inhibitory concentration (MIC) against phytopathogenic fungi. Results: Among the 20 strains, 90% were identified as Streptomyces and 10% as Kitasatospora. All the strains showed antagonisms against two or more of five phytopathogens: Corynespora cassiicola, Colletotrichum guaranicola, Colletotrichum sp., Pestalotiopsis sp., and Sclerotium coffeicola. Streptomyces spp. strains LaBMicrA B270 and B280 were active against phytopathogens of the guarana plant (Paullinia cupana). Furthermore, AcOEt/2-propanol 9:1 extract from the 10-day strain LaBMicrA B280 cultured medium presented activity against all the phytopathogens tested, with a minimum inhibitory concentration of 125 μg/mL. Conclusions: The results revealed various actinomycetes in three rhizospheres of I. edulis in the Amazon and the high potential of metabolic extracts from some of these bacterial strains against phytopathogenic fungi that destroy numerous crops.

Ganoderma sichuanense is a widely studied medicinal mushroom, but the production of its antioxidant proteins has been scarcely evaluated. We assess the influence of different concentrations of culture media components under submerged fermentation, with and without agitation, on production of proteins with antioxidant activity from the mycelial biomass of G. sichuanense. Protein extracts were characterized by scanning electron microscopy, X-ray diffraction, and attenuated total reflectance Fourier-transform infrared spectroscopy. They were also analyzed for total protein and phenolic contents, antioxidant activities (ABTS^•+, DPPH^•, chelating ability, and reducing power), and electrophoretic profiles by SDS-PAGE. The most active extract was tested for cytoprotective potential under H₂O₂-induced oxidative stress in Saccharomyces cerevisiae. Growth kinetics of the best fermentation condition were also analyzed. Microstructural differences ranged from fibrillar to aggregated forms, depending on cultivation. Crystallinity was unaffected, but chemical differences and secondary structure organization were confirmed by infrared spectroscopy. The extract from the static culture with 10 g·L⁻¹ glucose, 5 g·L⁻¹ yeast extract, and 2.5 g·L⁻¹ soy peptone (referred as CM1S) showed the highest protein and phenolic contents and the strongest antioxidant activity (IC₅₀ = 4.8 and 24.0 µg of protein·mL⁻¹ for ABTS^•+ and DPPH^•, respectively). SDS-PAGE revealed higher protein band intensities in static cultures. CM1S showed potential to protect yeast cells from oxidative stress. The Gompertz model estimated a specific growth rate of 0.0068 h⁻¹ in CM1S. The findings highlight a cultivation strategy that modulates fungal metabolism and improves the recovery of antioxidant proteins from G. sichuanense biomass.

Los hongos de podredumbre blanca tienen uno de los sistemas de degradación de lignina oxidativa más eficientes, y presentan un complejo enzimático que es responsable de digerir materia lignocelulósica. El objetivo de este estudio fue evaluar la producción de lacasa y peroxidasas totales por la Ganoderma lucidum a través de la fermentación de estado sólido, utilizando semillas de azaí y aserrín de aceituno (Simarouba amara) no suplementario y suplementado con trigo, maíz y salvado de arroz. G. lucidum se inoculó en sustratos preparados con residuos y se incubó a 25 ºC. Las extracciones enzimáticas se realizaron en los sustratos de cultivo cada 2 días durante 30 días. Las actividades enzimáticas se determinaron utilizando ABT (2,2′-azino-bis(3-etilbenzotiazolina-6-ácido sulfónico) sal diamonico), con la adición del cofactor enzimático H2O2 para peroxidasas totales. La actividad de lacasas fue mayor en los residuos suplementados, con énfasis en el sustrato a base de azaí el día 16 de cultivo, mientras que en aceituno la actividad máxima era el día 6. En el residuo de azaí no suplementado, el máximo pico de actividad fue el día 8 y, en el aceituno, no se observó un crecimiento fúngico. En cuanto a las peroxidasas totales, G. lucidum cultivado en los sustratos azaí suplementados mostró picos de actividad en el día 8, 12, 16 y 28, y el día 6 y 12 en condiciones no suplementadas. Mientras que en el aceituno, la actividad total de peroxidasas se observó solo el día 6 de cultivo. Por lo tanto, G. lucidum mostró potencial para producir lacasas y peroxidasas totales, con una suplementación con sustrato que induce la síntesis de estas enzimas. Os fungos da podridão branca possuem um dos sistemas de degradação da lignina oxidativa mais eficientes, e apresentam um complexo enzimático responsável pela digestão da matéria lignocelulósica. O objetivo deste trabalho foi avaliar a produção de lacase e peroxidases totais por Ganoderma lucidum, via fermentação em estado sólido, utilizando sementes de açaí e serradura de marupá (Simarouba amara) não suplementadas e suplementadas com farelo de trigo, milho e arroz. G. lucidum foi inoculado em substratos preparados com resíduos e incubado a 25 ºC. Extrações enzimáticas foram realizadas nos substratos de cultura a cada 2 dias durante 30 dias. As atividades enzimáticas foram determinadas utilizando-se o sal de diamônio ABTS (2,2'-azino-bis(ácido 3-etilbenzotiazolina-6-sulfônico), com a adição do cofator enzimático H2O2 para as peroxidases totais. A atividade de lacase foi mais elevada nos resíduos suplementados, com destaque para o substrato à base de açaí no 16o dia de cultivo, enquanto no marupá, a atividade máxima foi no 6o dia. No resíduo não suplementado de açaí, o pico máximo de atividade foi no 8º dia e, no marupá, não foi observado qualquer crescimento de fungos. Quanto às peroxidases totais, o G. lucidum cultivado nos substratos suplementados de açaí apresentou picos de atividade nos dias 8, 12, 16 e 28, e nos dias 6 e 12, em condições não suplementadas. Enquanto que no marupá, a atividade total da peroxidase foi observada apenas no 6o dia de cultivo. Assim, G. lucidum mostrou potencial para produzir lacases e peroxidases totais, com a suplementação de substrato induzindo a síntese dessas enzimas. White rot fungi have one of the most efficient oxidative lignin degradation systems, and present an enzymatic complex that is responsible for digesting lignocellulosic matter. The aim of this study was to evaluate the production of laccase and total peroxidases by Ganoderma lucidum via solid-state fermentation using açaí seeds and marupá (Simarouba amara) sawdust unsupplemented and supplemented with wheat, corn and rice bran. G. lucidum was inoculated in substrates prepared with the residues and incubated at 25 ºC. Enzymatic extractions were performed on the culture substrates every 2 days for 30 days. The enzymatic activities were determined using ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt), with the addition of the enzymatic cofactor H2O2 for total peroxidases. The laccase activity was higher in the supplemented residues, with emphasis on the açaí-based substrate on the 16th day of cultivation, while in marupá the maximum activity was on the 6th day. In the unsupplemented açaí residue, the maximum peak of activity was on the 8th day and, in marupá, no fungal growth was observed. As for total peroxidases, G. lucidum cultivated in the supplemented açaí substrates showed activity peaks on the 8th, 12th, 16th and 28th day, and on 6th and 12th day under unsupplemented conditions. While, in the marupá, total peroxidase activity was observed only on the 6th day of cultivation. Thus, G. lucidum showed potential for producing laccases and total peroxidases, with the substrate supplementation inducing the synthesis of these enzymes.