Mycoprotein represents an alternative protein source that offers both nutritional and health benefits. This study aimed to evaluate the production of mycoprotein from Lentinus squarrosulus through solid-state fermentation (SSF) using soybeans as the substrate. Fermentation was carried out for 15 days at 23−25 °C on both the control (soybean only) and soybean fermented with Lentinus squarrosulus. Parameters measured included biomass yield, proximate composition, amino acid profile, and antioxidant activity. Results showed that the fungus successfully colonised the substrate on day 10 (32.35 ± 2.54 g), followed by a decline at day 15, indicating an optimal fermentation window before nutrient depletion. SSF resulted in increased moisture (11.37%), ash (5.74%), and carbohydrate content (28.16%), whereas protein decreased from 44.76% to 36.43% compared with the control. Essential amino acids, including threonine, leucine, and phenylalanine, were reduced after fermentation. The antioxidant activity decreased in the fermented sample, with an IC₅₀ of 32.05 ± 1.01 µg/mL, compared to 25.10 ± 0.91 µg/mL in the control. These findings demonstrate that soybean-based SSF supports the growth of L. squarrosulus and biomass formation; however, optimisation is required to enhance protein quality and preserve bioactive components. Overall, L. squarrosulus remains a promising yet underutilised candidate for sustainable mycoprotein development.

Fusarium wilt, caused by Fusarium oxysporum f. sp. Lycopersici, is a major soil-borne disease threatening tomato (Solanum lycopersicum) production worldwide, causing yield losses of up to 80 %. This study evaluated the antifungal potential of the medicinal mushroom Lentinus squarrosulus against FOL, aiming to identify bioactive metabolites for sustainable disease management. FOL was isolated from infected tomato plants in Coimbatore, identified morphologically and molecularly (GenBank accession PQ350410) and used for in vitro assays. Nine macro basidiomycetes were screened via dual culture assay; L. squarrosulus TNAU L3 showed the highest mycelial growth inhibition (55.72 %), followed by Auricularia auricular (49.26 %) and L. squarrosulus TNAU L2 (43.74 %). Ethyl acetate extracts of L. squarrosulus significantly inhibited F. oxysporum f. sp. lycopersici growth in agar well diffusion assays, with inhibition increasing from 37.44 % at 500 ppm to 67.53 % at 2000 ppm. GC-MS analysis of extracts revealed antimicrobial compounds including eicosatrienoic acid, benzofuran, isopimaric acid and cyclododecanol, linked to antifungal and immunomodulatory activities. Pathway analysis indicated these metabolites are synthesized via the pentose phosphate pathway, contributing to both direct pathogen suppression and enhancement of plant defense through metabolic modulation. The dual mechanism-pathogen inhibition and host immunity activation-suggests L. squarrosulus as a promising eco-friendly biocontrol agent. This work highlights the potential of macrobasidiomycete-derived metabolites as alternatives to synthetic fungicides and provides a preliminary framework for developing natural fungicidal formulations. Further field validation and optimization of metabolite application are needed to advance integrated management strategies for Fusarium wilt in tomato.

ABSTRACT: Nanotechnology has emerged as a versatile field with applications across numerous disciplines. This study focuses on the biosynthesis of iron nanoparticles (FeNPs) using an aqueous extract of Lentinus squarrosulus. The formation of FeNPs was confirmed by a distinct color change at 60°C, followed by spectroscopic analyses, including ultraviolet-visible (UV-Vis) and Fourier-transform infrared (FTIR) spectroscopy. The antibacterial efficacy of FeNPs was assessed against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Streptococcus mutans, Enterococcus faecalis, and Enterobacter aerogenes, demonstrating significant inhibitory activity. Additionally, FeNPs exhibited notable photocatalytic degradation of Methylene Blue, Fast Green, and Congo Red dyes, highlighting their potential for wastewater treatment. The findings suggest that FeNPs synthesized via Lentinus squarrosulus could be valuable in biomedical applications and environmental remediation.

Bioremediation Potential of Lentinus squarrosulus (Mont.) Singer in Runoff Wastewater from Rivers State, Nigeria

Five white rot fungal species (Earliella scabrosa, Lentinus squarrosulus, Schizophyllum commune, Trametes sp.1, and Trametes sp.2) were evaluated for their ability to decolorize four synthetic textile dyes (Remazol Brilliant Blue R [RBBR], Reactive Orange 16, Reactive Black 5, and Amaranth) on agar plates supplemented with 50, 100, and 150 ppm of each dye. Decolorization zones were measured every 48 h across 14 days, and dye decolorization percentage was calculated using the Plate Volume Method (PVM). E. scabrosa decolorized all four dyes across the entire concentration range, although efficiency declined at high concentrations. L. squarrosulus decolorized RBBR and Reactive Black 5 dyes, completely removing RBBR even at 150 ppm, but achieving only 69% removal of Reactive Black 5 at that concentration. Trametes sp. 1 and 2 effectively decolorized RBBR by completely removing the dye even at 150 ppm, whereas S. commune exhibited no decolorization of any dye. Statistical analysis highlights potential differences in the characteristics or behaviors of these five fungal species, even when decolorizing the same textile dye. These findings identify E. scabrosa and L. squarrosulus as promising candidates for the biological treatment of dye-contaminated wastewater, supporting their potential scale-up in sustainable wastewater management solutions.

Unveiling the ethnomedicinal mystery of Lentinus squarrosulus Mont.: a multi-dimensional exploration of morphological, molecular profiling and optimization of cultural and nutritional conditions

Cosmeceutical industry is continually searching for new substances or extracts from natural sources that are nontoxic and can delay the aging process. Lentinus squarrosulus is a wild edible mushroom valued by local communities in Asian countries as food as well as its medicinal benefits. However, the bioactive substances and cosmeceutical biological properties of L. squarrosulus mycelia grown in submerged culture have not been demonstrated. This study aims to investigate the feasibility of producing L. squarrosulus mycelial biomass and cosmeceutical bioactive compounds through large-scale submerged fermentation. L. squarrosulus mushroom mycelia grown in a 5-L airlift bioreactor submerged culture was investigated as a new source of cosmeceutical biological substances. An ethanolic extract derived from L. squarrosulus mycelia possesses potential antioxidant agents capable of inhibiting dermal enzymes such as collagenase, elastase, and tyrosinase for simultaneously treating skin aging and hyperpigmentation. The Molisch test, FTIR analysis, and b-glucan assays revealed that the main polysaccharide present in the L. squarrosulus mycelial extract was b-glucan (64.3 %), which acts as a major contributor to its cosmeceutical properties. The L. squarrosulus mycelial extract was non-toxic to human skin keratinocytes (HaCaT). Cosmeceutical formulations developed with the mycelial extract, especially in essence form, preserved the extract’s bioactivities, presented a pale-yellow color and a pH of 6.0, which is considered suitable for cosmeceutical design. This study provides new information regarding submerged growth of L. squarrosulus mycelia in an airlift bioreactor, which has potential as a novel source of useful bioactive b-glucans for cosmeceutical applications. High mycelial biomass was produced over a short time in this airlift bioreactor that concomitantly had anti-aging and skin whitening properties. The results suggest that the mycelial extracts of L. squarrosulus could be further developed to produce safe natural ingredients for inclusion in cosmeceutical products. HIGHLIGHTS This is the first report detailing the submerged growth of Lentinus squarrosulus mushroom mycelia in an airlift bioreactor. L. squarrosulus mycelia grown submerged in an airlift bioreactor is a new source of b-glucan with multifunctional cosmeceutical properties. Ethanolic extracts of L. squarrosulus mycelia have the potential to simultaneously treat skin aging and hyperpigmentation. GRAPHICAL ABSTRACT

Cloning and expression of a short manganese peroxidase from Lentinus squarrosulus and its potential in enhancing the digestibility of crop residues.

The increasing problem of carbon dioxide emissions has become a significant concern, with mushroom production identified as one of the contributing factors. This is because the mushroom production process emits carbon dioxide through respiration, and the carbon dioxide emitted by the mushrooms contributes to an increase in greenhouse gases. The carbon dioxide emitted by mushrooms can be utilized in various applications, such as supporting vegetable cultivation in greenhouses. However, the amount of carbon dioxide emitted by mushrooms varies. Thus, this study aimed to investigate the growth, yield, and carbon dioxide emissions in five edible mushrooms, namely Pleurotus pulmonarius, Lentinus squarrosulus, P. ostreatus, P. citrinopileatus, and P. cystidiosus. The experiment found that the fresh weight, dry weight, and biological efficiency percentage of P. ostreatus were the highest at 65.71, 11.18 g, and 28.22 percent, respectively. In contrast, the fresh weight, dry weight, and biological efficiency percentage of L. squarrosulus were the lowest, at 24.90, 3.80 g, and 9.90 percent, respectively. On the other hand, the carbon dioxide emitted from L. squarrosulus was the highest, ranging from 854.00 to 8369.67 ppm, while the carbon dioxide emitted from P. cystidiosus was the lowest, ranging from 606.00 to 861.00 ppm.

Exploring the nutritional attributes of wild edible mushrooms can be vital in addressing global food security. However, there is a dearth of studies on wild edible mushrooms regarding their identification, nutritional status, and molecular characterization in Mizoram, North East India, an integral landscape of the Indo-Burma hotspot region. In the present study, samples of wild edible wood-growing mushrooms were collected from the Mamit and Champhai District of Mizoram. The present study investigated the identities and phylogenetic relationships of the eight fungal species using molecular approaches. The mushrooms were identified up to the species level based on fungal sequences with known identities in GenBank, viz. Auricularia delicata, Laetiporus sulphureus, Panus roseus, Lentinus squarrosulus, Pleurotus cystidiosus, Pleurotus djamor, Pleurotus pulmonarius and Schizophyllum commune. Further, the identified mushrooms were analysed for macro and micro-nutritional values to assess their suitability for human dietary intake. Results revealed that the collected wild edible mushrooms were rich sources of protein (17.79 - 36.46 g/100 g), carbohydrates (58.14 - 33.77 g/100 g) and fibres (5.63 - 8.32 g/100 g), while the amount of fat (1.93 - 2.77 g/100 g) was low. In addition to bio-molecules, the mushroom samples contained appreciable amounts of essential minerals. Therefore, the selected mushrooms can be used as a potential foodstuff and may also be used as a supplementary protein diet to maintain human health and help achieve food security and sustainable development goals. The present results encourage bio-prospecting, and the need to explore wild edible mushrooms further, especially in the least explored global biodiversity hotspots for sustainable livelihood.

In-silico screening and metabolomic profiling identify Naltrindole, a unique isoquinoline from Lentinus squarrosulus (Mont.,1842) mushroom having potent anti-diabetic properties

Bacteria membered in lactic acid producing group containing many dominant properties can be applied in diverse fields. This research was aimed to select effective lactic acid bacterial strains and enhanced their growth with mushroom extracts. Fermented vegetable samples were used as a source to achieve lactic acid bacteria by isolating on MRS medium. Fifteen isolates were lactic producing bacteria that exhibited clear zone around their colony on medium. Bacterial isolates FV2, FV4, and FV14 have produced the highest lactic acid content in quantitative investigation. In addition, FV2 and FV4 have been found the ability to suppress growth of pathogenic bacteria Salmonella Typhimurium and Staphylococcus aureus, with the highest inhibitory activity at 200 AU/mL. Isolates FV2 and FV4 have been classified as Lactiplantibacillus plantarum FV2 and Pediococcus pentosaceus FV4 based on their 16S rRNA gene sequences. Mushroom extracts of Pleurotus pulmonarius and Lentinus squarrosulus (Mont.) have been discovered oligosaccharide, and antioxidants activity, including the property to support the growth and lactic acid production of L. plantarum FV2 and P. pentosaceus FV4. High survival of lactic acid bacteria within bio−bead gel structure have been found more than naked cell under strong acid condition, including storage at 4 °C up to 28 days. Bio−bead gels containing lactic acid bacteria and mushroom extracts could be applied as the starter culture to produce fermented milk that they promoted coagulation of milk protein, high lactic acid bacteria, and antioxidant activity.

Coagulation and flocculation are integral basic unit processes for conventional water treatment plants. Usually, chemical coagulants are used in most of the treatment plants. However, the search for natural bio-coagulants is ongoing to reduce the negative impacts of chemical coagulants on human health and the environment. In this research, a natural bio-coagulant from a local fungus (Lentinus squarrosulus) was produced. The sedimentation process in river water by this myco-coagulant was investigated using a settling column. Kaolin suspension was used as synthetic turbid water for the settling column tests. Detention time and the overflow rate of the particles are necessary to design sedimentation basins for water treatment plants. As such, tests were conducted to plot the iso-removal lines for the kaolin particles. Such data is required to design sediment or settling basins for the water treatment facilities. Therefore, detention times and overflow rates of the kaolin particles were calculated for an optimum myco-coagulant dose of 1% (v/v). To reduce 80% of the initial turbidity from the kaolin suspension, the overflow rate and detention time of the sedimentation tank should be 41.6 m/day and 59.5 minutes, respectively. In contrast, similar ranges of overflow rates and detention times could remove only about 23% of the turbidity from the kaolin suspension without any myco-coagulant. This novel, natural and biodegradable coagulant is found to have the potential for reducing turbidity in river water; therefore, also can be a good candidate for the coagulation-flocculation process in water treatment plants.

Lentinus squarrosulus is a wild edible mushroom utilized globally not only for its nutritional values but also for its medicinal and mycoremediation potentials. Domestication of this mushroom will make available mother culture and spawn for research and propagation, and ensure year-round availability for economic and sustainable development. Tissue culture was prepared and actively growing mycelia were inoculated onto grain spawn. Cultivation trials were conducted using sawdust from various wood species, including Treculia africana (African breadfruit), Mangifera indica (Mango), Dacryodes edulis (African pear), and mixed sawdust from various wood species. Substrates were composted, sterilized, inoculated with the spawn of L. squarrosulus and incubated. Growth was monitored, recorded and fruiting bodies harvested. Results of domestication revealed that mother culture was produced in 5 - 7 days spawned in 14 days and was available for research, and cultivation. L. squarrosulus mycelia colonized all the substrates used to varied degrees with the mycelial running time ranging from 30.4 days to 34.8 days. Mycelia running time on T. africana differed significantly from D. edulis. Fruiting bodies were successfully harvested from 38 to 68 days with the maximum number of fruiting bodies (40 ± 9.47) and highest yield of 89.03 ± 29.41 g obtained from T. africana in three flushes. This was followed by M. indica (35, 54.27 ± 14.64 g). Dacryodes edulis sawdust recorded the lowest yield (23, 32.31 ± 11.34 g). M. indica sawdust had the broadest pileus diameter (6.45 ± 1.97 cm) and longest stipe (2.83 ± 0.49 cm). In conclusion, L. squarrosulus has the potential to be domesticated in Orlu, Imo State, with T. africana sawdust as a suitable substrate for cultivation.

In the present study, a unique mushroom belonging to the genus Lentinus was collected from the Lateritic region of West Bengal, India. The purpose of our investigation is to determine its taxonomic identity and evaluate its therapeutic properties, with a particular emphasis on its antioxidant and antimicrobial properties. A thorough morpho-molecular investigation revealed that the collected mushroom is a novel variety that has been named Lentinus squarrosulus var. odoratus in recognition of its potent, rotting banana-like aroma. The methanolic extract of the desiccated basidiocarps exhibits potent antioxidant properties, as evidenced by the EC50 values that range from 314.201 ± 3.12 to 933.48 ± 3.58 μg/mL, contingent upon the type of free radicals employed. The extracted fraction was also discovered to be effective in inhibiting the growth of human-pathogenic bacterial strains in a dose-responsive manner, as evidenced by the low MIC50 values, which varied from 281.61 ± 1.64 to 680.31 ± 4.12 μg/mL depending on the strains under investigation. The inhibitory effect was shown to be potentially caused by an increase in membrane porosity and damage to the cell wall, resulting in the leaking of intracellular components. The findings of this research indicate that there is a significant therapeutic potential encased in this mushroom, necessitating further comprehensive investigation.

The ethnomedicinal subtropical mushroom Lentinus squarrosulus Mont. is a polypore with high wood-degrading potential. This study aimed to collect L. squarrosulus isolates, analyze lignocellulolytic enzymes - Laccase (Lac), Lignin peroxidase (LiP) and Manganese peroxidase (MnP) and evaluate substrates for mass production. Sporophores of Lentinus sp. (Isolate L1 to L5) were collected from different regions of Tamil Nadu and characterized morphologically and molecularly. Qualitative screening tests with guaiacol substrate demonstrated the production of laccase by all the isolates; however, isolate L3 showed maximum reddish-brown zonation. Additionally, isolate L3 produced the highest green colouration on the ABTS (2,2’-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid) substrate. This confirmed the oxidation of ABTS to ABTS-azine in the presence of laccase, outperforming the other L. squarrosulus isolates. All the isolates tested with Azure-B agar for LiP and MnP showed faint discolouration, suggesting lower secretion of peroxidase enzymes. Quantitative analysis using a lignolysis basal medium also confirmed that isolate L3 secreted Lac predominantly, followed by LiP and MnP enzymes extracellularly. All isolates showed peak Lac activity at pH 4.5, while LiP and MnP production were highest at pH 5.0. The optimal temperature for all lignolytic enzymes was 28±2ºC. Maximum laccase secretion occurred between the 5th - 7th day after inoculation (DAI), with LiP and MnP peaking at 7 DAI. Nine agro-waste substrates were evaluated, with isolate L3 showing the highest biological efficiency on Paddy straw (99.62%), followed by Arecanut sheath (97.43%) and Groundnut shell (95.15%). This study enhances our understanding of L. squarrosulus enzymatic capabilities. It provides insights for mass production, which has strong potential for industrial bioremediation and waste management due to its high laccase production.

Mushrooms are eukaryotic organisms with spores, without chlorophyll that reproduce sexually and asexually, mushrooms can be used by humans as food ingredients and ingredients for traditional and modern medicine. The purpose of this study was to determine the type and habitat characteristic of macroscopic fungi conducted in the kateri procted forest area Malaka Regency. Using exploration and identification methods where the plant identification process is a process of matching a plant according to a particular taxonomi on each research transect line. There are 28 species of macroscopic fungi namely Dacrymyces stilatus, Auricularia auricula-judae, Xylaria hypoxylon, Daldinia concentrica, Microporus xanthopus, Earliella scabrosa, Pycnoporus sanguineus, Polyporus alveolaris, Lentinus squarrosulus, Hexagonia tenuis, Trametes pubescens, Tyromyces chioneus, Marasmius oreades, Marasmius haematocephalus, Baespora myosura, Marasmius rotula, Marasmius andorsaceus, Trogia infundibiliformis, Leucocoprinus brebissonii, Lepiota helveola. Agaricus micromegathus, Bolbitius vitelinus, Mycena albicocapilaris, Crepidotus variabilis, Parasola plicatilis, Coprinopsis fragilisimus, Lycoperdon pyriforme, Pleurotus djamor. Divided into 5 Orders namely Dacrymycetales, Auriculariales, Xylariales, Polyporales, Agaricales. And 12 family namely Dacrymycetaceae, Auriculariaceae, Xylariaceae, Polyporaceae, Marasmiaceae, Agaricaceae, Bolbitiaceae, Mycenaceae, Crepidotaceae, Psathyrellaceae, Schizophyllaceae, Pleurotaceae. The most abundant habitat Characteristics were on rotten wood branches as many 9 species, on dead wood stem many as 8 species, in leaf little many as 4 species and in soil with hums many as 7 species.

The physicochemical and microbial attributes of storm water samples prior to and after mycofiltration was determined using routine methods. The preparation of the substrate was done by supplementation of un-fermented sawdust with calcium carbonate, calcium sulphate, granulated sugar and wheat offal. The mixture was allowed to undergo composting for 7 days upon which, it was inoculated with Lentinus squarrolsulus spawn and incubated at room temperature. The sample was then passed through a network of mycelia for pollutant removal. The mean TDS and EC concentrations of the raw and mycofiltered samples was 1369.8 ± 4.5 and 516.4 ± 2.9 mg/l as well as 2785.0 ± 4.2 and 1251.0 ± 5.6 µS/cm. Pb, Cu, Cd, Cr and Fe readings for the raw samples were 0.03±0.03mg/l, 0.07±0.03 mg/l, 0.01±0.01mg/l, 0.03±0.01mg/l and 1.00±0.11mg/l. For the mycofiltered samples, the Pb, Cd and Cr were reduced to nil while Cu and Fe decreased to 0.03 ± 0.03 mg/l and 0.47± 0.09 mg/l. The difference between the mean trace metal values recorded for the raw and filtered samples was insignificant (p>0.05). The results indicated that mycofilter derived from L. squarrolsulus mycelia was capable of purifying storm water sample.

This study investigated protein hydrolysates obtained from Lentinus squarrosulus and Lentinus edodes fruiting bodies via gastric protease hydrolysis and ultrafiltration, yielding peptides with a molecular weight below 6.5kDa. These hydrolysates displayed significant tyrosinase inhibitory activity similar to positive controls, peptide from Chinese quince seed (RHAKF) and kojic acid. L. squarrosulus-derived hydrolysates exhibited superior antioxidant properties compared to L. edodes in DPPH (47% vs. 23%) and ABTS (77% vs. 23%) assays. Identified bioactive peptides, particularly LILGGSSS from L. squarrosulus, interacted with tyrosinase through hydrogen bonds at specific residues. Notably, these protein hydrolysates showcased potent tyrosinase inhibition without cytotoxic effects, presenting promising prospects for addressing hyperpigmentation caused by excessive tyrosinase activity from stress or UV exposure.

This study explores the potential of fungal mycelium-based plate-like composites as sustainable and cost-effective alternatives for synthetic materials. Locally available lignocellulosic waste and fungi isolated locally were used to fabricate mycelium composites. The medium primarily consists of Albizia sawdust and the fungi Lentinus squarrosulus and Pleurotus ostreatus were used. All the composites identified here were subjected to analyses based on scanning electron microscopy, moisture content, dry density, water absorption, flammability, thermal stability, and compression strength following ASTM and ISO standards. The mycelium plates produced with L. squarrosulus mycelium (ASL) exhibited lower dry density and less water absorption. The ASL mycelium plates exhibited a soft and foamy appearance, whereas the P. ostreatus material (ASP) appeared dense mycelium growth only in the top and bottom parts of the plate. The UL-94 rating demonstrated that both samples exhibited superior flame retardancy properties (rated as V1) compared to commercially used expanded polystyrene (EPS). These variations were due to the difference between mycelium density. Therefore, these new bio-composites have the potential to replace conventional packaging or interior construction materials.