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.

One of the most expensive steps in mushroom production is the disinfection of the substrate. This study aimed to evaluate different methods of disinfecting various substrates used in the cultivation of Pleurotus ostreatus from the Amazon. P. ostreatus was grown under uncontrolled temperature conditions in a greenhouse (at approximately 30 °C) on substrates formulated with residues (seeds) of açaí and tucumã palm trees, Brazil nuts shell and pine sawdust. The different substrate disinfection treatments comprised autoclaving, immersion in hot water or immersion in calcium hydroxide (2%). The chemical composition of residues, substrates, and mushrooms was analyzed. During cultivation, it was found that in the treatment with hot water, all the bags were contaminated, and treatment with calcium hydroxide inhibited the colonization and growth of the mushrooms. P. ostreatus grown on the autoclaved açaí substrate supplemented with tucumã had higher production. The highest percentage of fiber content was obtained when the mushrooms were cultivated in the pine substrate supplemented with Brazil nuts (32.68 to 33.65%), and the highest protein content (20.03 to 20.88%) was found in the mushrooms grown on açaí-based substrates. Considering that P. ostreatus showed higher productivity in autoclaved substrates, further studies should be performed to develop alternative methods of disinfecting substrates for the cultivation of mushrooms in the Amazon region, as well as, in other regions with a tropical climate. However, the cultivation of this mushroom is promising using some regional substrates, especially with açaí, which is one of the important natural products of the Amazon.

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.