Empowering health with mushroom biofortification: nutritional and medicinal attributes

Abstract

Undernutrition, sometimes known as “hidden hunger”, affect over two billion individuals globally, primarily due to low mineral content in staple food. Minerals play a very crucial roles in human health, influencing various functions from bone strength to heart rhythm regulation. Biofortification or enriching processed foods of staple crops have emerged as effective strategies to combat deficiencies, significantly enhancing mineralabsorption and bioavailability. In this context, mushrooms stand up as a potential candidate with their diverse species and exceptional ability to accumulate minerals and other bioactive ingredients, offering a valuable resource for enriching food. However, biofortification of edible mushrooms via addition of certain metals such as lithium, zinc, selenium, iron and others presents a promising sustainable approach for delivering essential nutrients to human being. Thus, continued research and optimization of biofortification methods hold promise for improving human health andcombating hidden hunger on a global scale.

Introduction

The prevalence of micronutrient deficiencies gradually spreads throughout the world, representing a widespread problem known as hidden hunger. This issue predominantly arises from the consumption of staple foods that lack essential minerals (von Grebmer et al. 2014). Recently several food industries are performing biosensor, artificial intelligence and different quality control technique for food security throughout the word. In coming days several natural based food products are also going through the system (Chhetri 2024; Meliana et al. 2024; Nath et al. 2024). Minerals play indispensable roles in physiological processes, encompassing bone integrity, nerve impulse transmission, and cardiac rhythm regulation, thus underscoring their paramount importance in sustaining optimal health and longevity. Key minerals contribute not only to hormonal synthesis but also to the maintenance of cardiac regularity. Macro- and micro-elements constitute integral constituents of dental (calcium, phosphorus, and fluorine) and skeletal (calcium, magnesium, manganese, boron, and iron) structures, while various micro-elements (Cu, manganese, Fe, Se, and Zn) serve as vital cofactors for numerous enzymatic reactions. Macro elements (calcium, phosphorus, magnesium, sodium and potassium) exert pivotal roles in neural processes, including signal transduction and transmission, in contrast, microelements are indispensable for erythrocyte production (cobalt [Co], I, and Fe), glucose homeostasis (chromium [Cr]), and antioxidant defense activation (molybdenum [Mo]) (Freeland et al. 2012; Singh et al. 2024). Fortifying processed foods with minerals represents an efficacious strategy to combat mineral deficiencies, augmenting absorption and bioavailability levels significantly. Biofortification, encompassing modern biotechnological approaches, stands out as a promising path for enhancing the nutrient profile of staple crops like common beans, maize, wheat, rice, cereals, and legumes, through the ultimate aim of ameliorating human health (Singh et al. 2016; Duggal et al. 2022). Notably, augmenting edible mushrooms through bio-enrichment has emerged as a compelling modality to deliver vital nutrients to the populace (Bertelsen 2013). The burgeoning commercial cultivation of various mushroom species, coupled with their adeptness in mineral bioaccumulation, renders mushroom a promising candidate for enriching dietary staples with essential minerals crucial for human health (Niedzielski et al. 2014). Human consumption of mushrooms spans centuries, driven not solely by their gustatory appeal but also by their nutritional prowess (Bertelsen 2013). Diverse organic compounds inherent to mushrooms, including glucans, phenolic compounds, and organic acids, confer beneficial effects on human health (Petrovic et al. 2015). Phenolic compounds, comprising phenolic acids, showcase antioxidative and anti-inflammatory attributes (Reis et al. 2012; Muszynska et al. 2013), while organic acids such as ketoglutaric, fumaric, succinic, malic, citric, and tartaric acids exhibit antioxidant properties and immune-stimulating capabilities, besides regulating metal ion availability through chelation, complexation, and adsorption phenomena (Çayan et al. 2014; Lee et al. 2007; Zhang et al. 2020). The profile and abundance of phenolic acids and organic acids exhibit considerable interspecies variation (Gąsecka et al. 2018). Ground mushroom powder finds utility in enhancing the nutritional composition of diverse foodstuffs, including snacks, burgers, cakes, pasta, and bread (Lu et al. 2020).

Biofortification, delineated by the World Health Organisation strives to heighten the dietary potency of nutrition crops via agronomical methodologies, traditional breeding practices, or recent biotechnological interventions (Sen et al. 2024). In contrast to conventional fortification entailing nutrient supplementation during processing, biofortification elevates crop nutrient levels during cultivation. Given the climatic variability, farmers are encouraged to tailor crop varieties to ensure optimal nutrient delivery (Porter et al. 2014). Fungi assume a pivotal ecological role as decomposers, facilitating organic matter degradation and nutrient release into the ecosystem (Gulis et al. 2019). Noteworthy advancements in mushroom biofortification have yielded fortified varieties, including closed -cup white and chestnut mushrooms, endowed with vitamin D and vitamin B12, thereby addressing deficiencies in these essential vitamins (Maity et al. 2023). The utilization of mushroom-incorporated food products has burgeoned owing to their medicinal attributes and nutritional merits. Various studies attest to the augmentation of protein, ash, and crude fibre content upon integrating powdered mushrooms into diverse food matrices, thereby enhancing their nutritional profile and quality. For example, adding 10% powdered mushroom to muffin and bread batter significantly increases the protein content compared to controls, improving the quality of the bread (Majeed et al. 2017). This review summarizes research on mushroom biofortification published from 2012 to 2024, highlighting the key developments, trends, and advancements in the field over this period. This review article synthesises existing literature while highlighting novel advancements in mushroom biofortification, including the use of Li, Se, and Zn, as well as the identification of new mushroom species and optimized cultivation methods. We have expanded on the bioavailability of these nutrients, emphasizing their neuroprotective and antimicrobial properties, which have been underexplored. Additionally, this review identifies critical gaps in scaling biofortification processes and the potential for industrial applications, contributing valuable insights to the field’s advancement and its implications for public health. For biofortification purposes, researchers have harnessed different mushroom species, each rich in distinct nutrients, as substrates. In summary, the convergence of biofortification strategies and mushroom bio-enrichment represents a promising frontier in addressing global malnutrition and hidden hunger. By harnessing the innate capacity of mushrooms to accumulate essential minerals and bioactive compounds, coupled with innovative agricultural practices, we can fortify staple crops and food products, thereby enhancing the dietary status or overall health outcomes of populations worldwide. Continued research and implementation efforts in this realm hold immense potential for combating nutrient deficiencies and promoting public health on a global scale.

Chief metals employed in biofortification

Fe, Zn, Se and Li are mostly utilised in mushroom biofortification process and also vital for human health. Metal ions can have a major effect on cellular functions in fungal cells by blocking important enzymes, interfering with biosynthesis, and changing metabolic pathways. For instance, too many metal ions can bind to active sites of enzymes or cause oxidative stress, which can disrupt enzyme activity and impair growth and metabolism. Optimising biofortification strategies while reducing potential toxic effects on beneficial fungal communities requires an understanding of these interactions (Detail explained in Table 1). Literature studies evident that metal speciifcally Se is incorporated as selenoproteins and selenoenzymes within the fruiting bodies (Kora 2020; Xu et al. 2021). However, it is also note that combined effect of different metals resulted in more nutrient content than the single dosage (Dhiman et al. 2024). Li, a naturally occurring alkali metal, is ingested through diet and exists in trace volumes in the human body. It is important for nourishing bodily functions, it contributes significantly to metabolism, neural signalling, and cell growth (Birch 2012). Recently Ogidi et al. (2023) demonstrated an edible mushroom; Pleurotus pulmonarius fortified with Cu, Li and Zn showed enhanced nutrient contents and bioactivities such as antioxidant as well as antimicrobial potentials.

Table 1.

Different functions of metal ions within fungal cells

Metal ion	Role in fungal cells	Enzyme inhibition	Biosynthesis processes	Metabolic pathways	Relevance to biofortification	References
Se	Antioxidant activity Enhances stress tolerance Improves nutrient content	Excess selenium can inhibit glutathione reductase	Boosts synthesis of antioxidant compounds like glutathione and selenoproteins Enhances polysaccharide and phenolic production	Reduces ROS through antioxidant pathways Influences sulfur metabolism due to its similarity with sulfur	Increases selenium content in mushrooms Improves nutritional value and antioxidant properties for human health	Hossain et al. (2021); Danso et al. (2023)
Zn	Essential cofactor for many enzymes (e.g., zinc metalloenzymes) Regulates protein synthesis and growth	High zinc levels can inhibit enzymes like urease and SOD through metal overload	Promotes synthesis of proteins, lipids, and nucleic acids	Regulates carbohydrate metabolism Involved in glycolysis and nitrogen metabolism	Improves protein and nutrient content Enhances enzymatic activity in mushrooms	Zięba et al. (2020); Koç and Karayiğit (2022)
Fe	Central to electron transport and redox reactions Vital for fungal respiration	Excess iron can lead to Fenton reactions, causing oxidative damage Can inhibit critical antioxidant enzymes like catalase in high concentrations	Supports heme and siderophore biosynthesis Enhances the production of bioactive compounds	Facilitates energy production through mitochondrial pathways Affects the tricarboxylic acid (TCA) cycle	Enhances nutritional value by increasing iron bioavailability in mushrooms	Pecoraro et al. (2021); Budzyńska et al. (2022)
Li	Modulates stress responses and growth Can affect ion balance in cells	Can inhibit enzymes such as glycogen synthase kinase-3 (GSK-3) and phosphatases, disrupting signal transduction pathways	May influence biosynthesis of secondary metabolites and stress	Affects signaling pathways such as inositol metabolism Impacts osmotic and ionic stress pathways	Potentially introduces lithium as a micronutrient Enhances stress resilience in fungal cells, supporting growth under adverse conditions	Van (2017); Jakobsson et al. (2017); Assunção et al. (2012)

Health impact

Studies reveal that dietary Li deficiency can cause severe psychological problems in humans and developmental problems in animals, highlighting the essential role that Li plays in mood regulation. Prolonged use may help meet the body’s need for Li, which may be increased by certain mood disorders. These days, personality disorders like manic depression are serious mental health issues. A vital and successful treatment for unipolar depression, acute mania, and bipolar affective disorder, the Li cation lowers aggression and suicidal thoughts ((Bauer and Gitlin 2016; Malhi and Outhred 2016).

Mechanisms of action

The mechanism of action of Li is probably through competing with similar cations, such as Na⁺ and Mg²⁺, to inhibit enzymes, hormones, and vitamins, thereby influencing intracellular processes and neurotransmission pathways. This may change how neurotransmitters involved in cellular metabolism are synthesised and released (Vosahlikova and Svoboda 2016). Li is involved in decreasing the expression of proapoptotic gene and antiapoptotic genes and protecting toxins. Furthermore, Li facilitates phosphorylation of GSK-3 and activates BMALI protein that further corresponds with neuroprotective effect and mood stabilization. The possible biochemical mechanisms underlying Li’s benefits are illustrated in Fig. 1.

Fig.1.

Illustrating the biochemical mechanisms of lithium's beneficial effects in humans

Selenium

Selenium is a crucial mineral certainly present in many foods or also existing as a nutritional supplement. Se is a constituent of 25 selenoproteins, such as thioredoxin reductases, glutathione peroxidases, and selenoprotein P. Selenoproteins are crucial for DNA synthesis, thyroid hormone metabolism, reproduction, and defence against oxidative damage and contamination (Lei et al. 2022).

Health impact

Se’s roles in apoptosis, DNA repair, the endocrine and immune systems, and its antioxidant properties suggest it may help prevent cancer (Rayman 2012). Animal studies also indicate Se’s potential chemopreventive properties. Additionally, selenoproteins reduce inflammation, prevent lipid oxidation, and inhibit platelet aggregation (Cold et al. 2015).

Mechanisms of action

Experimental studies on various cell types have shown highly variable results regarding Se, with effects ranging from anti-apoptotic to pro-apoptotic. However, nutritional study in rats showed that eating broccoli causes cardioprotection against ischaemia and reperfusion by activating survival proteins like Akt and Bcl-2 and generating an anti-apoptotic signal (Fig. 2). Later broccoli contains high levels of Se, it is reasonable to suggest that this cardioprotective effect is partially due to Se. Further support for Se’s anti-apoptotic effect comes from tests on animals using various selenoprotein suppression models (Tanguy et al. 2012).

Fig.2.

Cardioprotective benefits of intake selenium at physiological levels. GPx, glutathione peroxidase; TrxR, thioredoxin reductase; Sel S, selenoprotein, Bcl-2, B-cell lymphoma-2; GSH/GSSG, reduced/oxidized glutathione ratio; NFkB, nuclear factor kappa-B; AP-1, activator protein-1; TyrK, tyrosine kinase

Zinc

For many physiological processes, including newborn development, reproductive health, immune system performance, and vision, Zn is a necessary micronutrient. Because it is so important in many biological processes in the human body (Natasha et al. 2022).

Health impact

Zn is an essential mineral for biological systems because it detoxifies dangerous oxygen-free radicals, supports protein synthesis and gene expression, and preserves the structural and functional integrity of biological membranes (Jomova et al. 2022). ZnPoor growth, cognitive decline, prenatal problems, and elevated morbidity and mortality can result from a Zn deficiency (Karenya 2023). Deficiencies in Zn in humans can result in reduced appetite, anaemia, growing retardation, hypogonadism with diminished reproductive capacity, depressed mental function, and, in certain situations, impaired keratosis and teratogenic effects (Noulas et al. 2018).

Mechanisms of action

Numerous studies have been conducted on Zn’s role in inhibiting oxidative stress, emphasising the metal’s significance as an antioxidant and co-factor for metallothionein proteins. Zn limits NADPH oxidase and lowers high sugar-induced swelling in semipermeable membranes when combined with Fe and Cu. In order to protect cells from ROS SOD needs Zn to convert superoxide radicals into H₂O_₂ and oxygen. Adequate Zn levels are essential for antioxidant function, as deficiency impairs SOD and induces chronic endoplasmic reticulum stress, disrupting protein synthesis and Zn transport. Zn also inhibits free radicals through glutamate cysteine ligase and regulates cellular glutathione via Nrf₂. Zn is vital for metallothionein under normal conditions and is released during oxidative stress, enhancing metallothionein expression and anti-inflammatory effects. Zn levels control transporters (ZnTs and ZIPs) and are regulated by MTF-1 in response to cytoplasmic Zn²⁺ levels, with Zn stored in organelles like mitochondria and the endoplasmic reticulum (Hussain et al. 2022). (Fig. 3).

Fig.3.

Zinc’s role in regulating cellular oxidative stress and antioxidant activity. Nrf2, nuclear factor erythroid 2–related factor 2; GPX, glutathione peroxidases; H₂O₂, hydeogen peroxide; SOD, superoxide dismutase; MT, metallothionein; OH, Hydroxyl radical; NF-KB, nuclear factor kappa B; AP-1, activator protein 1

Iron

Earth contains iron, a transition metal and trace element. For all living things, it is an essential component. Iron limits environmental growth despite being abundant in the earth’s crust (Purves et al. 2012).

Health impact

For almost all living things, iron is essential. It is involved in many different metabolic processes, including the transport of oxygen, the synthesis of DNA, and the movement of electrons. But because iron can produce free radicals, the amount of iron in body tissues needs to be carefully controlled. There is a chance that high iron levels will damage tissue. Iron metabolism disorders are common in humans and include an extensive spectrum of conditions with different clinical presentations, from iron overload to anaemia, and possibly even neurodegenerative diseases (Abbaspour et al. 2014). The absence of mobilisable iron reserves and the presence of markers indicating an insufficient iron supply to tissues, such as the erythron, are the hallmarks of iron deficiency (Wick et al. 2013). Iron deficiency during pregnancy has been associated with a number of unfavourable outcomes, including an advanced risk of sepsis, perinatal or maternal mortality, and low birth weight. Moreover, anaemia and iron deficiency are linked to higher rates of morbidity and can have a detrimental effect on cognitive function (Abbaspour et al. 2014).

Mechanism of action

On the apical side of enterocytes, dietary iron is absorbed via DMT1 and DCytB. Together with hephaestin (HEPH), ferroportin (FPN1) can export excess iron from the cytosol to the bloodstream or store it in ferritin. The liver and spleen store excess iron, which circulates bound to transferrin and is mainly utilised by erythroid cells. HEPH, ceruloplasmin (CP), and zyklopen (ZP) assist FPN1 in exporting Fe³⁺ from cells. Under low iron conditions, the expression of DCytB, DMT1, and FPN1 increases due to the regulation of dietary iron absorption by hypoxia-inducible factor 2α (HIF2α). HIF2α inhibitors have the ability to reduce iron overload in conditions like hereditary hemochromatosis. The ferritin H (FTH) and L (FTL) subunits combine to form a "labile iron pool" that is stored in ferritin when there is an excess of intracellular iron. Because FTH possesses ferroxidase activity, Fe₂O can be stored as Fe₃O. When there is a cellular iron shortage, ferritinophagy is used by NCOA4 to release the iron that has been transferred to ferritin by chaperones PCBP1 and 2 (Silvestri et al. 2023). (Fig. 4).

Fig.4.

Control of the body’s various organs’ absorption and utilisation of iron. Fe³⁺, ferric iron; Fe²⁺, Ferrous iron; DMT1, Divalent metal transporter 1; Dcytb, duodenal cytochrome b; PCBP1-2, Poly(rC)-binding protein 1; NCOA4, nuclear receptor coactivator 4; FPN, ferroportin, Heph, hephaestin

Methods of biofortification in mushrooms

Substrate enrichment

The nutritional value of mushrooms is increased during cultivation by adding nutrients to the substrate, also known as the growth medium (Singhal et al. 2019). Minerals like Zn or Se, for example, can be added to the compost or casing soil so that mushrooms can grow and absorb these vital nutrients. For biofortifying button mushrooms (Agaricus bisporus) and oyster mushrooms (Pleurotus spp.), this technique is especially popular and successful because it guarantees steady nutrient uptake throughout the growth cycle, this method offers a sustainable and effective way to produce nutrient-dense mushrooms by enriching the substrate (Maknali et al. 2021; Tang et al. 2023).

Nutrient solution irrigation

The nutritional profile of mushrooms can be improved by irrigating them with nutrient-rich solutions while they are growing. By precisely controlling the kind and quantity of nutrients that the mushrooms absorb, this technique guarantees targeted fortification (Sangeeta et al. 2024). Iodine is necessary for thyroid health, and applying iodine-enriched solutions, for instance, helps fortify mushrooms with it. It is a useful method for creating nutrient-enhanced mushrooms because it is productive and flexible (Singh et al. 2022).

Genetic biofortification

Modern genetic methods are used to create strains of mushrooms that are better at absorbing or synthesising nutrients. In mushrooms, for example, altering metabolic pathways can promote the buildup of vital minerals like Zn or Se. Although this method has a lot of potential for creating nutrient-enriched mushrooms, it frequently encounters legal issues and public mistrust in some areas, which may prevent it from being widely used (Słyszyk et al. 2024).

Spraying nutrient solutions on fruiting bodies

A quick and easy way to increase the nutritional value of mushroom fruiting bodies is to spray nutrient solutions directly onto their surface. By ensuring that the nutrients stick to the mushrooms, this method enriches them without materially changing their growth environment. It is a useful method for targeted biofortification that enables accurate nutrient delivery (Cotter 2015).

Biofortification with different metals

Mushrooms can be a promising natural food for adding minerals that are necessary for human health because more and more species are being grown for commercial use with their exceptional bioaccumulation abilities (Rzymski et al. 2016). Numerous metals with nutritional and pharmacological significance can bioaccumulate in mushrooms (Table 2). (Budzyńska et al. 2021).

Table 2.

A list of metals used for biofortification of some edible mushrooms

Mushroom species	Substrate used	Additional supplement	Metal used	Concentration	References
P. ostreatus	Barley, rice straw substrate	Addition of millet mushroom flour in order to make porridge blended with zinc dense mushroom	Zn	50 mg/kg 100 mg/kg 200mg/kg 400mg/kg 600mg/kg	Zieba et al. (2020)
A. bisporus	Wheat straw (1000kg), chicken manure (900kg), gypsum (85kg)	Zinc nitrate hexahydrate dissolved with water	Zn	0.1, 0.2, 0.4, 0.6, 0.8, mmolL−1	Rzymski et al. (2017)
Pleurotus eryngii,	Mixture of beech and alder sawdust (1:1)	20% wheat bran with 5% cornmeal, 5% soymeal and 1% gymsum with 1% saccharose	Fe	(a)5mM	Budzyńska et al. (2022)
Pleurotus ostreatus				(b)10mM
Pholiota nameko				(c)50Mm
Agaricus bisporus	Straw	chicken manure, gypsum	Li	1.0, 5.0, 10, 50, 100, 500 mg kg−1 dw	Pankavec et al. (2021)
Ganoderma lucidum, Pleurotus eryngii	Mixture of beech and alder sawdust	wheat bran 20%, corn flour 5%, soybean meal 3%, sucrose 1% and gypsum 1%	Li	0.25, 0.5, 0.75 and 1.0 mM	Mleczek et al (2017)
Pleurotus ostreatus	Mixture of beech and alder sawdust (1:1)	wheat bran 20%, corn flour 5%, soybean meal 3%, sucrose 1% and gypsum 1%	Li	0.25, 0.5, 0.75 and 1.0 mM
Agrocybe cylindracea	Mixture of beech and alder sawdust (1:1)	20%, wheat bran, 10% wheat straw chaff, 5% corn flour, 3% soybean meal, 1% chalk and 1% gypsum	Li	0.25, 0.5, 0.75 and 1.0 mM	Rzymski et al. (2017)
Hericium erinaceus	Mixture of beech and alder sawdust	20% wheat bran, 5% corn flour, 3% soybean meal, 1% sucrose and 1% gypsum	Li	0.25, 0.5, 0.75 and 1.0 mM
P. holiota nameko	Mixture of alder and beech sawdust (1:1)	20% wheat bran and 5% cornmeal	Ca	5, 10, and 15 mM of Fe and 5 and 10 mM	Budzyńska et al. (2021)
P. floridanus	Sawdust	Wheat bran	Ca	1:1, 1:5, 1:10, and 2:1 (in ratio)	Odiketa et al. (2020)

Li

In commercially grown edible mushrooms, Li concentrations are generally observed to be very low. While most vegetables and grains are recognised for their richness in Li, certain mushroom varieties such as Craterellus cornucopioides, Amanita strobiliformis, and Pleurotus ostreatus are known to contain varying concentrations of this element. The Li-enriched substrates that edible mushrooms take up nutrients from have a direct impact on the amount of Li present in those mushrooms. Several studies have shown that Li from mushrooms is more readily available and easier to access to for the patients than psychiatric medications (de Assunção et al. 2012).

According to Mleczek et al. (2017) if the substrates were fortified with 1 mM of Li₂CO₃ (or 0.5 mM for P. eryngii), consuming a meal comprising 100 g of dry Ganoderma lucidum, Pleurotus ostreatus first flush, P. ostreatus second flush, and P. eryngii would provide 7.4, 1.2, 1.6, and 1.9 mg of Li, respectively. The similar number of mushrooms full-grown on substrates supplemented with 1 mM of CH₃COOLi, on the other hand, would result in Li intakes of 2.6 mg for G. lucidum, 0.8 mg for P. ostreatus first flush, 0.5 mg for P. ostreatus second flush, and 1.5 mg for P. eryngii. The concentrations of Li accumulated in the fruiting bodies of the mushrooms, however, were not thought to be high enough for direct psychiatric treatments, though they might be supportive of daily Li intake for general health or behavioural modification. In a study by De Assunção et al. (2012). Pleurotus ostreatus mushrooms were grown on coffee bean husk enriched with Li at concentrations of up to 500 mg per kg. This resulted in a substantial increase in the fruiting bodies’ Li concentration by a factor of 2–5. Furthermore, in vitro digestion testing revealed that 70% of the Li in the enriched mushrooms was bioaccessible. Similar observation was also noticed by Vieira et al. (2013) who reported that P. ostreatus grown on coffee husk substrate enriched with 500 mg lithium chloride per kg could accumulate 39.5 mg of Li per kg of dry weight.

According to Rzymski et al. (2017), if 100 g of the fresh biomass of Agrocybe cylidracea and Hericium erinaceus is consumed, they may be able to provide 21–24% and 69%, respectively, of the Recommended Dietary Allowance (RDA) for Li when grown by 0.75 mM Li acetate or 1 mM Li chloride. But the amounts of Li found in the fruiting bodies of these mushrooms were not high enough to justify their use in mental treatments, which usually require doses between 600 and 1200 mg daily.

Faria et al. (2019) found that when Lentinus crinitus, a basidiomycete, was grown on a malted extract medium with 25 mg/L of Li (Li), it accumulated Li up to 575 mg/g. However, this high accumulation was associated with reduced mycelial biomass. To match the Li intake from a 300 mg Li carbonate pill (56 mg Li), one would need to consume 97 g of Li-enriched mycelial biomass. This suggests that Li-enriched mushrooms have potential for meeting daily Li requirements, especially in psychiatric therapies. Optimising cultivation methods for popular mushroom species is essential for achieving effective Li concentrations in the fruiting bodies.

Faria et al. (2018) evaluated lithium bioaccumulation in P. ostreatus mycelia grown in a liquid malt extract medium fortified with Li₂CO₃ or LiCl. At 40 mg/L dose, the maximum lithium accumulation was noted. Lithium from both sources was efficiently bioaccumulated by P. ostreatus, but the accumulation was greater for Li₂CO₃ supplementation. These results demonstrated the potential of fungal biomass for biotechnological uses, such as alternative drug delivery methods and products enriched with lithium.

In a recent study Pankavec et al. (2021) investigated the Li improvement of white Agaricus bisporus mushrooms. They achieved this by fortifying the marketable growing substrate with Li₂CO₃ solutions at different concentrations of 1.0–500 mg per kg of dry weight. The results of the study showed that levels up to 100 mg kg⁻¹ dw showed a significant (p < 0.01) dose-dependent increase in Li accumulation in mushrooms. However, Li uptake in fruiting bodies was shown to decrease with increased fortification levels, saturating at 100 mg kg⁻¹. In addition, compared to mushrooms grown on control substrate, lithiated mushrooms display Li contents up to 300 times higher. The consequences of this study reveal the potential of lithiated mushrooms as a pro-medical substitute for Li salt-based treatments.

Therefore, on the basis of these observations it could be suggested a bioavailable and possibly advantageous natural Li source is provided by certain edible mushrooms which could be enhaced through varying the substrate compositions enriched with the metal. Li-enriched mushrooms may support daily intake, though they are insufficient for psychiatric treatment. Their pro-medical potential is further enhanced by optimised cultivation methods.

Se

The majority of cultivated edible mushrooms have very low Se contents (< 1–8.5 μg Se/g dry weight), making them Se deficient (Costa-Silva et al. 2011). The Se (Se) content in wild edible fungi and mushrooms ranges from 12 to 200 μg/g, but most wild species are unsuitable for large-scale cultivation and have limited culinary value. To address this issue, mushrooms can be biofortified with Se on substrates enriched with inorganic or organic Se forms (Kora 2020). Biofortification impacts mushroom growth and metabolism, with selenomethionine (SeMet) being the most effective substrate. Hericium erinaceus is particularly effective in converting inorganic Se into organic Se proteins like Se-polysaccharides and seleno-AA. Se-biofortified mushrooms have shown potential in reducing tumour incidence, inhibiting cancer cell growth, and serving as dietary supplements and nutraceuticals (Maity et al. 2023).

Flammulina velutipes, also set an another example of biofortified mushroom through Se biofortification using selenite. After treatment with 20 μg/g of selenite, the mushroom accumulated 108 μg/g of carbon-based Se, with over 97% being organic. Most of the Se was bound to proteins (60–74%) and polysaccharides (15–21%), primarily as selenomethionine (56.8%), selenocysteine (22.8%), and methylselenocysteine (17.3%). Se biofortification also increased biomass yield and improved the mushroom’s content of proteins, polysaccharides, amino acids, and minerals like Ca, Fe, and Cu, making it a potential Se supplement (Dong et al. 2021).

Researchers examined the response of the medicinal fungus Inonotus hispidus to sodium selenite. The fungus tolerated up to 3.85 mmol/L selenite, which caused 85% growth inhibition, with 50% inhibition occurring at 1 mmol/L. Mycelium grown in 0.29 mmol/L Se had a Se content 115 times higher than that of control media, with 86% of it being organic. Microscopic observations showed that higher Se stress caused hyphal shrinkage, deformation, and collapse of clamp connections, though conidial production increased. Upon returning to standard media, mycelial growth was recovered. Se accumulation did not significantly affect the yield of Se-containing compounds but improved mycelial quality by reducing crude fat and ash content (Song et al. 2022).

By transforming inorganic Se into bioavailable organic forms like selenomethionine, Se biofortification improves the nutritional profile of edible mushrooms. Biofortified mushrooms exhibit promise as dietary supplements and nutraceuticals with anticancer and health-promoting qualities while also increasing Se content and biomass.

Zn

Zn supplementation and food fortification are recommended to reduce childhood infectious diseases in Zn-deficient populations. A study explored the potential of finger millet mixed with Zn-biofortified oyster mushroom powder to improve Zn bio-accessibility. Oyster mushrooms were grown on straw with zinc sulphate, dried, ground, and mixed with finger millet at different ratios. Zn content increased with higher zinc sulphate concentrations, with the 20% mushroom blend showing the highest bioaccessible Zn. However, this ratio negatively affected taste and aroma. The study concluded that lower mushroom powder ratios provide a balance between improved Zn levels and sensory appeal (Karenya 2023).

Koreti et al. (2023) studied the submerged culture of Calocybe indica with Zn-supplemented medium to enhance exopolysaccharide (EPS) production. After 21 days, Zn fortification increased both EPS and mycelial biomass, with the highest EPS yield (0.3853 ± 0.006 g/L) at 175 mg/L Zn concentration and maximum mycelial biomass (7.7133 ± 0.30 g/L). The study demonstrated that Zn supplementation boosts mineral content, influencing mycelial growth and EPS production. These bioactive EPS have potential applications in pharmaceuticals and food products.

Zięba et al. (2020) investigated the ability of P. eryngii mycelia from in vitro cultures and fruiting bodies to accumulate Zn and Se. They evaluated the efficiency, chemical composition, and bio-element content following biofortification with zinc sulphate, zinc hydro-aspartate, and sodium selenite (50 mg/L), each providing 20 mg/L of Zn ions. Phenolic compounds and lovastatin were detected using reverse-phase HPLC. The study found that P. eryngii effectively accumulated Se and Zn, with the type of Zn salt influencing the amount of Zn absorbed.

According to research by Oyetayo et al. (2021), Pleurotus pulmonarius grown on ZnSO₄-fortified substrates had the highest Zn content (349.5 mg/100 g) during harvest.

The nutritional value of edible mushrooms is increased through Zn biofortification, which also increases exopolysaccharide production with possible food and pharmaceutical uses and promotes Zn bio-accessibility. While the type of Zn source has a significant impact on absorption efficiency, optimal fortification strikes a balance between increased Zn levels and sensory appeal.

Iron

Iron (Fe) fortification of mushrooms can alleviate global deficiencies; however, the factors influencing the effectiveness of fortification remain incompletely understood. Three mushroom species (P. ostreatus, P. eryngii, and Pholiota nameko) were used in a study to compare the effects of three different forms of iron (FeSO₄ 7H₂O, FeCl₃ 6H₂O, and FeHBED) at three different concentrations (5, 10, or 50 mM) on the chemical composition, phenolic compounds, and organic acid production. For metal growth in all species, the 50 mM concentration worked best. P. eryngii and P. nameko benefited most from FeCl₃ 6H₂O, which increased Fe content by up to 145% and 185%, respectively, in comparison to controls. FeHBED increased the Fe content by 108% and was most effective for P. ostreatus. The highest Fe accumulation was found in P. nameko (89.2 ± 7.51 mg/kg DW). Fe salt supplementation generally inhibited phenolic acid production, except in P. ostreatus at 5 mM FeCl3 6H₂O and in P. eryngii at 5 mM FeSO₄7H₂O and FeCl₃6H₂O. Further, organic acid was found to be impacted by Fe supplementation (Budzyńska et al. 2022).

Scheid et al. (2020) assessed the bioavailability of iron in the mycelial biomass of various edible and medicinal basidiomycetes, including Plurotus ostreatus, P. eryngii, Ganoderma lucidum, and others, cultivated in sugarcane molasses and malt extract media. Iron content in the mycelial biomass ranged from 106–213 mg/kg in malt extract and 358–1304 mg/kg in sugarcane molasses. Biomass grown on molasses had significantly higher iron levels (1.68–6.84 times) than that on malt extract. P. ostreatus grown on molasses had the highest iron content (1304 mg/kg), 12.5 times higher than common iron-rich foods like broccoli and spinach. Significant species-specific differences were observed (p < 0.001) across fungi.

The effectiveness of iron fortification varies by species, iron form, and concentration, but it greatly increases the iron content of mushrooms. The highest iron accumulation was seen in P. nameko and P. ostreatus, with sugarcane molasses serving as an excellent growth medium.

Nutritional analysis of biofortified mushrooms

In a biofortification study published in 2018 by Rathore et al. demonstrated, Calocybe indica’s response to Se-enriched wheat straw. According to the study, inorganic Se was accumulating within the fruit bodies and integrating into proteins (56–68%), polysaccharides (22–29%), and nucleic acids (1.4–2.7%). Harvested from the substrate enriched with 10 mg/ml Se, fruit bodies had the highest protein content (25.31 g/100 g). At a concentration of 10 mg/ml Se, amino acid profiling showed that the highest concentrations of Glu (4.73 g/100 g) were tracked by Asp (1.80 g/100 g) and glycine (1.61 g/100 g).

Kaur et al. (2018) examined the application of Se-rich wheat straw for biofortification in P. florida, P. sajor-caju, and P. ostreatus. According to the study, fruiting bodies treated with Se showed an increase in their overall protein content. The protein content of P. florida increased significantly, from 1.644 mg/g in the control to 2.492 mg/g in the samples treated with Se. When P. ostreatus was treated with Se, its total protein content rose from 1.728 mg/g in the control to 1.786 mg/g. P. sajor-caju also increased with Se biofortification, going from 2.496 mg/g in the control to 2.640 mg/g.

In 2019, Fasoranti et al. investigated Se-fortified Pleurotus species, including P. ostreatus and P. pulmonarius. The Se-fortified mushrooms exhibited higher protein content (16.35–18.21%), while the carbohydrate content remained unchanged (53.84–58.10%) compared to non-fortified samples. Both fortified and unfortified mushrooms had low fat content (0.39–0.40%). Calcium was the most abundant mineral (1.78–20.94 mg/100 g), and nickel was the least (0.01 to 0.03 mg/100 g), with no lead detected. Glu was the predominant amino acid (12.27 mg/100 g), and both essential and non-essential amino acids were present in all samples. Interestingly, Fadugba et al. (2024) showed Se-fortified P. ostreatus had highest fat, ash and crude fiber content than the non-fortified one justifying the utilization of the fortified mushroom as a nutrient rich food for human consumption.

Hu et al. (2019) investigated Auricularia auricular Se-enriched fruiting bodies, finding consistently high protein and polysaccharide levels, with Se treatment not significantly altering these amounts. However, at 40 μg/g selenite, essential, non-essential, and total AA decreased, with alanine being the most abundant. Similarly, Ogidi and Oyebode (2023) examined the proximate composition and mineral content of Pleurotus pulmonarius mushrooms, fortified with trace elements. Cu and Li fortifications increased protein and ash content, while Zn fortification raised protein (up to 17.90%) and reduced carbohydrates. The Zn + Cu + Li combination had the highest protein (18.93%) and ash (8.30%), significantly altering the nutritional profile (p ≤ 0.05).

Rathore et al. (2020) conducted research on the fortification of C. indica with Vitamin D2 through artificial (UVB light) and natural (sunlight) methods. The effects of the treatment on the nutritional value of the treated mushrooms were analysed. Their investigation revealed that the highest amount of Vitamin D2 was present in mushrooms treated with sunlight, which was 78.33 μg/g, while those treated with UVB light had reached a maximum of 140.58 μg/g.

Se and trace element biofortification enhance the protein, amino acid, and mineral content of mushrooms, improving their nutritional value. Vitamin D2 enrichment further boosts their dietary significance, offering a potential solution to nutrient deficiencies.

Biological activities

Edible mushrooms offer a wide range of healthy benefits as they are already enriched with various metabolites as well as biologically active macronutrients (Aytar and Ozmen 2020a, 2020b, 2020c)). In recent days, bio-fortifications come up with a new avenue to explore fully their medicinal properties. Several reports have shown biofortified mushrooms expressed enhanced bioactivities compared to unfortified one. However, the most significant activities observed are antioxidant activity followed by antimicrobial, anticancer and immunological activity. Literature survey revealed that a good source of organic Se was obtained from aqueous enzymatic extracts of Se-enriched white button mushrooms (A. bisporus) grown on Se-rich compost. Without changing the organoleptic qualities of food, these extracts provide a practical and repeatable supplement for reaching the daily recommended intake of Se. While avoiding the risks associated with excessive Se intake, the addition of these Se-rich extracts to meals may help to prevent and treat diseases like neurodegenerative, cardiovascular, and immunological disorders that are linked to low Se levels (Cremades et al. 2012). Furthermore, the enriched mushrooms support their role in health and disease management by enhancing their antimicrobial, antioxidant, anticancer, and immunological properties.

Antioxidant activity

The antioxidative activity of fortified mushrooms aligns with Vieira et al. (2013), who studied Pleurotus ostreatus cultivated on coffee husk supplemented with Zn, Li, and iron. Using the DPPH antioxidant test, significant differences (p < 0.05) were found between treatments, though no direct relationship was observed between total phenolic content and antioxidant activity, differing from previous research. Variations in antioxidant activity were attributed to the bioavailability of enriched elements. In the β-carotene-linoleic acid system, the enriched and non-enriched P. ostreatus extracts showed no significant differences. Overall, antioxidant activity ranged from 58 to 66%, indicating the high antioxidative potential of P. ostreatus.

de Souza Lopes et al. (2022) showed mushrooms bioaccumulated with Li salts grew on coffee husks could oxidise substances. According to their findings, P. ostreatus and P. djamor that had been enriched with Li exhibited increased oxidising activity against catalase and SOD, with the exception of the highest Li salt concentration. Li has several health benefits, including the preclusion and treatment of neurodegenerative ailments, which may account for the increased antioxidant activity of Li-enriched mushrooms. Li has psychopharmacological, neuroprotective, and prophylactic benefits in preventing mood disorders when taken with food (Szklarska and Rzymski 2019).

Recently, in 2024, Wang et al. conducted a unique strategy to enhance the antioxidant activity of P. eryngii polysaccharides through biofortification with different selenium salts such as sodium selenite and selenium yeast. Results had shown that biofortication made changes on molecular weight and molar ratios of compositional monomers of the polysaccharides extracted from fortified mushrooms. Selenium yeast Se-enriched P. eryngii polysaccharides showed enhanced radical scavenging activity than sodium selenite Se-enriched polysaccharides indicating different fortification process impacted the structural chemistry of the bioactive molecules along with their biological activities.

Fortified mushrooms exhibit high antioxidant activity, influenced by bioavailable elements like Li, which also provide neuroprotective and mood-stabilising benefits. This highlights their potential in promoting overall health and preventing neurodegenerative disorders (Table 3).

Table 3.

Antioxidant property of different biofortified mushroom species

Mushroom species	Antioxidant property	Key findings	References
P. sajor-kaju	DPPH radical scavenging activity, metal chelating activity, lipid peroxidation	The total antioxidant activity of methanol extracts from Se-enriched mushrooms increased with concentration, showing significantly higher antioxidant content compared to non-enriched controls i.e., 4.58 mg BHT/g DW, significantly higher than the control’s 1.45 mg BHT/g DW,	Bhatia et al. (2014)
P. florida, P. ostreatus, and P. sajor-caju	A higher DPPH scavenging activity. The scavenging effect was expressed as scavenging activity (1 2 Ac/Ad),	Protein samples demonstrated varying degrees of antioxidant activity, as indicated by their DPPH radical scavenging effects. Samples with lower absorbance readings at 515 nm exhibited higher DPPH scavenging activity, suggesting stronger antioxidant properties	Kaur et al., (2018)
G. frondosa	The DPPH radical-scavenging activity	The DPPH radical-scavenging activity of the aqueous samples increased with rising concentrations. Higher concentrations showed greater scavenging rates	Li et al. (2017)
C. indica	DPPH, Ferric reducing antioxidant power	Se biofortification significantly enhanced the antioxidant properties of C. indica fruit bodies, with 5 mg/ml Se increasing total phenolic compounds to 25.29 mg GAE/g and improving DPPH radical scavenging and FRAP activity. The highest IC50 value for DPPH was found in non-supplemented mushrooms, indicating lower antioxidant activity compared to Se-enriched samples	Rathore et al. (2018)
Cordyceps militaris	Scavenging Activity of DPPH Radicals	The DPPH radical scavenging activity of both water and ethanol extracts from Se-biofortified C. militaris was significantly higher than that of control extracts, with water extracts showing superior activity (88.5%–96%) compared to ethanol extracts (33.3%–90.4%). The scavenging activity of ethanol extracts increased with concentration	Hu et al. (2019)
Coriolus versicolor	Scavenging ability on DPPH radicals, Chelating abilities on ferrous ions	The methanolic extracts demonstrated significant, concentration dependent antioxidant activities, evidenced by enhanced DPPH radical scavenging, effective ferrous ion chelation, and increased reducing power. These effects were comparable to standard antioxidants, including ascorbic acid and α-tocopherol, indicating the extracts’ potential in mitigating oxidative stress	Miletić et al. (2019)
PPS – Pleurotus pulmonarius (fortified with selenium) POS – Pleurotus ostreatus (fortified with selenium	DPPH free radical scavenging activity, Hydroxyl radical scavenging ability, Reducing power assay	Selenium-enriched Pleurotus spp. demonstrated significantly greater reductive and hydroxyl radical scavenging abilities compared to non-enriched counterparts, with both activities being concentration-dependent. At 250 µg/mL, the hydroxyl scavenging activity reached 91.58% for POS and 88.48% for PPS	Fasoranti et al. (2019)
Volvariella volvacea	DPPH radical scavenging, metal chelating, and lipid peroxidation inhibition activities	Se-enriched mushroom extracts demonstrated significantly higher antioxidant activity compared to controls, with 71.1% DPPH radical scavenging at 10 mg/ml. Additionally, Se-enriched exhibited superior metal chelation power of 56.1%, suggesting that selenium enrichment enhances both radical scavenging and metal chelation abilities	Kora (2020)
Button, oyster, and paddy straw mushroom		When compared to unfortified mushrooms, the antioxidant action was improved in terms of total phenolics, total antioxidant, DPPH radical scavenging, metal chelating, and lipid peroxidation inhibition activities
Hericium erinaceum	DPPH assay and reducing power assay	Polysaccharides, particularly selenium-containing ones, exhibited strong dose-dependent antioxidant activity, reaching 100% at 5 mg/mL, surpassing control polysaccharides (72%) and common antioxidants like BHT and α-tocopherol. Se-enriched polysaccharides showed the highest antioxidant activity, with low EC50 values (< 0.1 mg/mL) indicating their superior efficacy
Pleurotus spp. such as P. citrinopileatus, P. djamor, and P. pulmonarius	The antioxidant activity was determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical (Sigma-Aldrich, St. Louis, MO, USA)	The mycelium of P. pulmonarius showed the highest total phenolic content (259 mg/100 g d.w.) and antioxidant activity (19% DPPH reduction) among the species under study. Although Pleurotus djamor mycelium demonstrated strong antioxidant potential in both gastric (25.16%) and intestinal juice (10.11%), Pleurotus citrinopileatus fruiting bodies demonstrated the highest antioxidant activity in artificial intestinal juice (1.18% DPPH reduction). These findings show that antioxidant capacity varies by species under different biological condition	Włodarczyk et al. (2020)
Pleurotus eryngii	DPPH radical scavenging activity	The DPPH scavenging activity of Pleurotus eryngii fruiting bodies significantly increased with selenium enrichment, indicating its potential as a dietary antioxidant. Selenium-enriched mushrooms showed notably higher antioxidant activity than non-enriched species, likely due to the incorporation of selenium into their polysaccharide structures, enhancing their hydrogen atom-donating capacity	Zięba et al. (2020)
Pholiota nameko	A free radical scavenger with metal chelating ability	Mushrooms’ antioxidant activity is linked to hydrogen donation and radical scavenging, but P. nameko lacks sufficient organic acids for effectiveness	Budzyńska et al. (2021)
P. pulmonarius	DPPH assay, nitric oxide (NO) assay	Fortifying mushrooms with zinc or iron did not significantly affect the yield or nutrient content, while zinc-fortified mushrooms exhibiting the highest protein content and superior scavenging activity against DPPH (96.3%) and NO (96.8%) radicals, comparable to butylated hydroxytoluene (97.1%). Simultaneous Zn and Fe supplementation increased ash content and enhanced the phenolic content	Oyetayo et al. (2021)

Antimicrobial activity

The use of Se-rich wheat straw for the biofortification of three Pleurotus species, P. ostreatus, P. florida, and P. sajor-caju, was examined by Kaur et al. (2018). Their research revealed that these species’ crude extracts and proteins exhibited antibacterial activity against a variety of pathogens, including Gram-positive (S. aureus) and Gram-negative (E. coli, A. hydrophila, K. pneumonia, Y. enterocolitica, and S. typhi) ones. It’s interesting to note that the antimicrobial qualities were not considerably changed by Se enrichment.

According to Mudroňová et al. (2019), ZnSO₄·7H₂O concentrations ranging from 100–500 mg increased Lactiplantibacillus plantarum’s lactic acid production, facilitated its adhesion to porcine enterocytes, and inhibited pathogens. Since Cu promotes the growth rate of lactic acid bacteria, bio-enrichment of edible mushrooms with Cu is encouraged.

Ogidi and Oyebode. (2023) used agar well diffusion to test the antimicrobial activity of Cu- Li, and Zn fortified P. pulmonarius against a range of microorganisms. Against Klebsiella pneumoniae, the ethanolic extract from mushrooms fortified with Cu showed the largest zone of inhibition, followed by extracts from P. pulmonarius fortified with Li, Cu + Li, and Cu + Zn + Li. Furthermore, against Klebsiella oxytoca and Candida tropicalis, the P. pulmonarius extract fortified with Zn, Cu, and Li exhibited the strongest antimicrobial activity.

Zn, Cu, Li, and Se-fortified mushrooms have stronger antibacterial activity, particularly against Candida and Klebsiella species. Potential as functional foods with health-promoting qualities is presented by these fortified mushrooms.

Anticancerous activity

Multiple stages of cancer, including initiation, promotion, and progression, can be modulated and blocked by Se supplementation, according to extensive research and literature reviews. Se-fortified button mushrooms (A. bisporus) were shown to produce strong anticarcinogens in female Sprague–Dawley rats. DMBA-induced mammary epithelial DNA adducts were decreased, and liver and mammary GSH were markedly activated upon enrichment with Se-biofortified mushrooms. With studies demonstrating increased cancer cells activity in Se-fortified mushrooms compared to inorganic forms, whereas treatment with Se-enriched mushrooms effectively inhibited tumour growth, notably inhibiting cell proliferation more than unfortified mushrooms (Bhatia et al. 2014).

Furthermore, by preventing lymphangiogenesis, Li biofortification has demonstrated promise in preventing colon cancer metastases. TGFBIp is less effective in colon cancer cells because it prevents Smad3 phosphorylation through GSK3beta inactivation. It had been noted that TGFBIp-driven lymphatic endothelial cell migration in tumour cells was inhibited by Li. While Li effectively inhibited lymphangiogenesis in tumours. However, it had no effect on the growth of SW620 tumours in vivo or in vitro. Based on tumour xenograft models, Li was found to inhibit TGFBIp-induced tumour lymphangiogenesis, which in turn regulates metastasis in the liver, lymph nodes, and lungs. In conclusion, Li suppressed TGFBIp-induced lymphangiogenesis in early tumour growth by acutely inhibiting TGFBIp expression, preventing colon cancer metastases (Pattanayak and Das 2022).

Immunological activity

The effects of mycelial water extracts of Lentinula edodes containing Zn and Se on the expression of CD4 + and CD8 + T cells stimulated with anti-CD3 antibody were examined by Kałucka et al. (2023). ^The results showed that the percentage of CD4 + CD69 + T cells was significantly downregulated in all examined extracts. It is noteworthy that, in comparison to the control, the non enriched extracts showed the strongest effect. Despite the lack of a statistically significant impact on CD25 expression on CD4 + T cells (p > 0.05), the non-enriched extract showed a tendency towards downregulation. Moreover, no extract had any effect on CD8 + T cell expression of the CD69 marker (p > 0.05); on the other hand, only the Zn and Se non-enriched fraction markedly increased the proportion of CD8 + CD25 + T cells. Thus, it could be suggested that Zn and Se enrichment modulated the immunological activity whereas their mechanisms of action appear to be different.

Practical challenges and implications of biofortification

Scalability of biofortification

Large-scale production of biofortified crops, such as mushrooms, poses difficulties in terms of resource management, infrastructure development, and cultivation method optimisation in order to satisfy demand worldwide. Scalability must be attained in order for biofortification to be successfully applied globally (Jangra et al. 2024).

Consumer acceptance

Adoption of biofortified products by consumers is critical to their success. In order to promote the inclusion of biofortified foods in regular diets and increase awareness of their nutritional advantages, public education campaigns are crucial. To achieve broad acceptance, it will be essential to dispel doubt and misunderstandings (Razzaq et al. 2021; Sandhu et al. 2023).

Cost-effectiveness

The sustainability of biofortified crops depends on the cost-effectiveness of biofortification techniques for both producers and consumers. Making these solutions accessible and affordable requires striking a balance between the possible health benefits and the cost of biofortification techniques, especially in areas with high rates of nutritional deficiencies (Ofori et al. 2022; Sandhu et al. 2023).

Food biofortification with minerals

Mineral deficiencies can be addressed safely, effectively, and economically with bio-fortification, especially through the bio-enrichment of mushrooms. A promising nutritional treatment for macro- and micronutrient deficiencies is the biofortification of mushrooms with elements such as Li, iron, Se, and Zn (Oyetayo 2023). Because of their well-known capacity to absorb and retain minerals, mushrooms are perfect for biofortification. Furthermore, MT which binds metals and promotes their accumulation, is found in mushrooms (Krezel et al. 2017). Commercial varieties of mushrooms typically have a low Se content, but growing them on substrates high in Se can increase this content. With chelated forms of Zn having lower phytotoxicity, Zn bio-accessibility and acceptance among local consumers has demonstrated promise in enhancing immune functions, particularly in preventing viral infections like COVID-19 (Li et al. 2017; Zhang et al. 2018).

Toxicity risks and mineral interactions

Although Se is necessary, too much of it can be toxic and cause symptoms like neurological problems and gastrointestinal distress (Ullah et al. 2023). Furthermore, deficiencies in vital nutrients like calcium and magnesium may result from high concentrations of one mineral blocking the absorption of others (Kiani et al. 2022). Consumers may be at risk from excessive Se accumulation in mushrooms, especially when they are grown in substrates high in Se (Guangzhi et al. 2021).

Conclusion

Biofortification of mushrooms represents a promising and feasible solution to address widespread micronutrient deficiencies and combat hidden hunger on a global scale. A significant body of research highlights the potential of mushrooms to accumulate essential nutrients, such as Li, Se, Zn, and iron, from enriched substrates, enhancing their nutritional value and broadening their applications as functional foods. Li-enriched mushrooms provide a bioavailable source of Li that can support daily intake, while Se biofortification increases the nutritional profile by transforming inorganic Se into bioavailable organic forms, such as selenomethionine, with anticancer and health-promoting benefits. Zn and iron biofortification further enhance the nutritional value of mushrooms, improving bio accessibility and addressing global deficiencies in these critical minerals. Additionally, fortified mushrooms, enriched with trace elements and vitamin D2, demonstrate potential in promoting overall health, preventing neurodegenerative disorders, and exhibiting antioxidant and antibacterial properties.

To transition from research to real-world applications, collaboration among policymakers, agricultural experts, and the food industry is essential to scale up production, optimize biofortification methods, and ensure equitable access to these nutrient-rich foods. Public awareness campaigns are also vital for driving demand and acceptance of biofortified mushrooms as part of a balanced diet. Continued research, coupled with innovative agricultural practices, holds great promise for improving global health outcomes, combating nutrient deficiencies, and enhancing public well-being. Investing in biofortification strategies will be a crucial step toward advancing global nutrition and addressing the challenges of hidden hunger.

Abbreviations

AA

Amino acids

Asp

Aspartic acid

Cu

Copper

GSH

Glutathione S-transferase

Glu

Glutamic acid

Li

Lithium

MT

Metallothionein

Mo

Molybdenum

ROS

Reactive oxygen species

Se

Selenium

SOD

Superoxide dismutase

TGFBIp

Transforming growth factor-beta-induced protein

Zn

Zinc

Author contributions

All authors contributed significantly to the manuscript. Monika Choudhary was responsible for collecting literature sources and drafting the manuscript. Parteek Mandyal critically reviewed the manuscript and contributed to the preparation of figures and tables. Anamika Chauhan assisted with table preparation and organized the references. Sandipta Ghosh provided supervision and was involved in writing, reviewing, and editing the manuscript.

Funding

There is no funder to acknowledge.

Data availability

The data will be available on request bases.

Code availability

Not applicable in this manuscript.

Declarations

Conflicts of interest

The authors declare that there are no conflicts of interest.

Ethical approval

Not applicable.

Consent to participate

Not applicable in this study.

Consent for publication

All authors have consent for publication.