Light is perceived by photoreceptors in fungi and further integrated into the stress-activated MAPK HOG pathway, and thereby potentially activates the expression of genes for stress responses. This indicates that the precise control of light conditions can likely improve the conidial yield and stress resistance to guarantee the low cost and long shelf life of Trichoderma-based biocontrol agents and biofertilizers. In this study, effects of wavelengths and intensities of light on conidial yield and stress tolerance to osmotic, oxidative and pH stresses in Trichoderma guizhouense were investigated. We found that 2 μmol photons/(m² × s) of blue light increased the conidial yield more than 1000 folds as compared to dark condition and simultaneously enhanced conidial stress resistance. The enhanced conidial stress resistance is probably due to the upregulated stress-related genes in blue light, which is under the control of the blue light receptor BLR1 and the MAP kinase HOG1. Full article

Background: MicroRNA plays an important role in multifarious biological processes by regulating their corresponding target genes. However, the biological function and regulatory mechanism of fungal microRNA-like RNAs (milRNAs) remain poorly understood. Methods: In this study, combined with deep sequencing and bioinformatics analysis, milRNAs and their targets from Trichoderma guizhouence NJAU 4742 were isolated and identified under solid-state fermentation (SSF) by using rice straw as the sole carbon source at 28 °C and 37 °C, respectively. Results: A critical milRNA, TGA1_S04_31828 (Tr-milRNA1), was highly expressed under heat stress (37 °C) and adaptively regulated lignocellulase secretion. Overexpression of Tr-milRNA1 (OE-Tr-milRNA1) did not affect vegetative growth, but significantly increased lignocellulose utilization under heat stress. Based on the bioinformatics analysis and qPCR validation, a target of Tr-milRNA1 was identified as Trvip36, a lectin-type cargo receptor. The expression of Tr-milRNA1 and Trvip36 showed a divergent trend under SSF when the temperature was increased from 28 °C to 37 °C. In addition, the expression of Trvip36 was suppressed significantly in Tr-milRNA1 overexpression strain (OE-Tr-milRNA1). Compared with the wild type, deletion of Trvip36 (ΔTrvip36) significantly improved the secretion of lignocellulases by reducing the retention of lignocellulases in the ER under heat stress. Conclusions: Tr-milRNA1 from NJAU 4742 improved lignocellulose utilization under heat stress by regulating the expression of the corresponding target gene Trvip36. These findings might open avenues for exploring the mechanism of lignocellulase secretion in filamentous fungi. Full article

Bicarbonate (HCO₃⁻) transporter family including the anion exchanger (AE) group is involved in multiple physiological processes through regulating acid-base homeostasis. HCO₃⁻ transporters have been extensively studied in mammals, but fungal homologues of AE are poorly understood. Here, we characterized the AE group member (MoAE4) in Magnaporthe oryzae. MoAE4 exhibits more sequence and structure homologies with the reported AE4 and BOR1 proteins. In addition to the common sublocalization on cytomembrane, MoAE4 also localizes on tonoplast. Yeast complementation verified that MoAE4 rescues boron sensitivity and endows NaHCO₃ tolerance in the BOR1 deleted yeast. MoAE4 gene is bicarbonate induced in M. oryzae; and loss of MoAE4 (ΔMoAE4) resulted in mycelial growth inhibited by NaHCO₃. Lucigenin fluorescence quenching assay confirmed that ΔMoAE4 accumulated less HCO₃⁻ in vacuole and more HCO₃⁻ in cytosol, revealing a real role of MoAE4 in bicarbonate transport. ΔMoAE4 was defective in conidiation, appressorium formation, and pathogenicity. More H₂O₂ was detected to be accumulated in ΔMoAE4 mycelia and infected rice cells. Summarily, our data delineate a cytomembrane and tonoplast located HCO₃⁻ transporter, which is required for development and pathogenicity in M. oryzae, and revealing a potential drug target for blast disease control. Full article

In this study, for the first time, the potential of rose flowers and lavender straw waste biomass was studied as feeding lignocellulose substrates for the cultivation of newly isolated in Bulgaria Ganoderma resinaceum GA1M with the objective of obtaining mycelium-based bio-composites. The chemical characterization and Fourier Transform Infrared (FTIR) spectroscopy established that the proximate composition of steam distilled lavender straw (SDLS) and hexane extracted rose flowers (HERF) was a serious prerequisite supporting the self-growth of mycelium bio-materials with improved antibacterial and aromatic properties. The basic physico-mechanical properties of the developed bio-composites were determined. The apparent density of the mycelium HERF-based bio-composites (462 kg/m³) was higher than that of the SDLS-based bio-composite (347 kg/m³) and both were much denser than expanded polystyren (EPS), lighter than medium-density fiber board (MDF) and oriented strand board (OSB) and similar to hempcrete. The preliminary testing of their compressive behavior revealed that the compressive resistance of SDLS-based bio-composite was 718 kPa, while for HERF-based bio-composite it was 1029 kPa and both values are similar to the compressive strength of hempcrete with similar apparent density. Water absorbance analysis showed, that both mycelium HERF- and SDLS-based bio-composites were hydrophilic and further investigations are needed to limit the hydrophilicity of the lignocellulose fibers, to tune the density and to improve compressive resistance. Full article

Solid state fermentation (SsF) is recognized as a suitable process for the production of enzymes using organic residues as substrates. However, only a few studies have integrated an evaluation of the feasibility of applying enzymes produced by SsF into subsequent hydrolyses followed by the production of target compounds, e.g., lactic acid (LA), through submerged-liquid fermentations (SmF). In this study, wheat bran (WB) was used as the substrate for the production of enzymes via SsF by Aspergillus awamori DSM No. 63272. Following optimization, cellulase and glucoamylase activities were 73.63 ± 5.47 FPU/g_ds and 107.10 ± 2.63 U/g_db after 7 days and 5 days of fermentation, respectively. Enzymes were then used for the hydrolysis of the organic fraction of municipal solid waste (OFMSW). During hydrolysis, glucose increased considerably with a final value of 19.77 ± 1.56 g/L. Subsequently, hydrolysates were fermented in SmF by Bacillus coagulans A166 increasing the LA concentration by 15.59 g/L. The data reported in this study provides an example of how SsF and SmF technologies can be combined for the valorization of WB and OFMSW. Full article

The present work highlights the valorization of the bulky recalcitrant lignocellulose byproduct wheat straw (WS) for the enhanced production of value-added xylanase by the locally sourced novel Penicillium chrysogenum strain A3 DSM105774 for the first time. The optimized production of xylanase by submerged state of fermentation of WS was achieved using a three-step statistical and sequential approach: one factor at a time (OFAT), Plackett–Burman design (PBD), and Box Behnken design (BBD). Incubation temperature (30 °C), WS, and ammonium sulphate were the key determinants prompting xylanase production; inferred from OFAT. The WS concentration (%(w/v)), yeast extract concentration (%(w/v)), and initial pH of the production medium imposed significant effects (p ≤ 0.05) on the produced xylanase, realized from PBD. The predicted levels of WS concentration, initial pH of the production medium, and yeast extract concentration provoking the ultimate xylanase levels (53.7 U/mL) with an 8.95-fold enhancement, localized by the estimated ridge of the steepest ascent of the ridge analysis path, were 3.8% (w/v), 5.1, and 0.098% (w/v), respectively; 94.7% lab validation. The current data underpin the up-scaling of xylanase production using this eco-friendly, cheap, and robust methodology for the valorization of WS into the value-added product xylanase. Full article

This study aimed to select thermotolerant yeast for bioethanol production from cellulose-rich corncob (CRC) residue. An effective yeast strain was identified as Saccharomyces cerevisiae TC-5. Bioethanol production from CRC residue via separate hydrolysis and fermentation (SHF), simultaneous saccharification and fermentation (SSF), and prehydrolysis-SSF (pre-SSF) using this strain were examined at 35–42 °C compared with the use of commercial S. cerevisiae. Temperatures up to 40 °C did not affect ethanol production by TC-5. The ethanol concentration obtained via the commercial S. cerevisiae decreased with increasing temperatures. The highest bioethanol concentrations obtained via SHF, SSF, and pre-SSF at 35–40 °C of strain TC-5 were not significantly different (20.13–21.64 g/L). The SSF process, with the highest ethanol productivity (0.291 g/L/h), was chosen to study the effect of solid loading at 40 °C. A CRC level of 12.5% (w/v) via fed-batch SSF resulted in the highest ethanol concentrations of 38.23 g/L. Thereafter, bioethanol production via fed-batch SSF with 12.5% (w/v) CRC was performed in 5-L bioreactor. The maximum ethanol concentration and ethanol productivity values were 31.96 g/L and 0.222 g/L/h, respectively. The thermotolerant S. cerevisiae TC-5 is promising yeast for bioethanol production under elevated temperatures via SSF and the use of second-generation substrates. Full article

As β-glucosidases represent the major bottleneck for the industrial degradation of plant biomass, great efforts are being devoted to discover both novel and robust versions of these enzymes, as well as to develop efficient and inexpensive ways to produce them. In this work, raw glycerol from chemical production of biodiesel was tested as carbon source for the fungus Talaromyces amestolkiae with the aim of producing enzyme β-glucosidase-enriched cocktails. Approximately 11 U/mL β-glucosidase was detected in these cultures, constituting the major cellulolytic activity. Proteomic analysis showed BGL-3 as the most abundant protein and the main β-glucosidase. This crude enzyme was successfully used to supplement a basal commercial cellulolytic cocktail (Celluclast 1.5 L) for saccharification of pretreated wheat straw, corroborating that even hardly exploitable industrial wastes, such as glycerol, can be used as secondary raw materials to produce valuable enzymatic preparations in a framework of the circular economy. Full article

Horizontal gene transfer, a process through which an organism acquires genes from other organisms, is a rare evolutionary event in yeasts. Artificial random gene transfer can emerge as a valuable tool in yeast bioengineering to investigate the background of complex phenotypes, such as heat tolerance. In this study, a cDNA library was constructed from the mRNA of a methylotrophic yeast, Ogataea polymorpha, and then introduced into Saccharomyces cerevisiae. Ogataea polymorpha was selected because it is one of the most heat-tolerant species among yeasts. Screening of S. cerevisiae populations expressing O. polymorpha genes at high temperatures identified 59 O. polymorpha genes that contribute to heat tolerance. Gene enrichment analysis indicated that certain S. cerevisiae functions, including protein synthesis, were highly temperature-sensitive. Additionally, the results confirmed that heat tolerance in yeast is a complex phenotype dependent on multiple quantitative loci. Random gene transfer would be a useful tool for future bioengineering studies on yeasts. Full article

Alternaria is a ubiquitous fungal genus in many ecosystems, consisting of species and strains that can be saprophytic, endophytic, or pathogenic to plants or animals, including humans. Alternaria species can produce a variety of secondary metabolites (SMs), especially low molecular weight toxins. Based on the characteristics of host plant susceptibility or resistance to the toxin, Alternaria phytotoxins are classified into host-selective toxins (HSTs) and non-host-selective toxins (NHSTs). These Alternaria toxins exhibit a variety of biological activities such as phytotoxic, cytotoxic, and antimicrobial properties. Generally, HSTs are toxic to host plants and can cause severe economic losses. Some NHSTs such as alternariol, altenariol methyl-ether, and altertoxins also show high cytotoxic and mutagenic activities in the exposed human or other vertebrate species. Thus, Alternaria toxins are meaningful for drug and pesticide development. For example, AAL-toxin, maculosin, tentoxin, and tenuazonic acid have potential to be developed as bioherbicides due to their excellent herbicidal activity. Like altersolanol A, bostrycin, and brefeldin A, they exhibit anticancer activity, and ATX V shows high activity to inhibit the HIV-1 virus. This review focuses on the classification, chemical structure, occurrence, bioactivity, and biosynthesis of the major Alternaria phytotoxins, including 30 HSTs and 50 NHSTs discovered to date. Full article

With the increasing world population, demand for industrialization has also increased to fulfill humans’ living standards. Fungi are considered a source of essential constituents to produce the biocatalytic enzymes, including amylases, proteases, lipases, and cellulases that contain broad-spectrum industrial and emerging applications. The present review discussed the origin, nature, mechanism of action, emerging aspects of genetic engineering for designing novel proteases, genome editing of fungal strains through CRISPR technology, present challenges and future recommendations of fungal proteases. The emerging evidence revealed that fungal proteases show a protective role to many environmental exposures and discovered that an imbalance of protease inhibitors and proteases in the epithelial barriers leads to the protection of chronic eosinophilic airway inflammation. Moreover, mitoproteases recently were found to execute intense proteolytic processes that are crucial for mitochondrial integrity and homeostasis function, including mitochondrial biogenesis, protein synthesis, and apoptosis. The emerging evidence revealed that CRISPR/Cas9 technology had been successfully developed in various filamentous fungi and higher fungi for editing of specific genes. In addition to medical importance, fungal proteases are extensively used in different industries such as foods to prepare butter, fruits, juices, and cheese, and to increase their shelf life. It is concluded that hydrolysis of proteins in industries is one of the most significant applications of fungal enzymes that led to massive usage of proteomics. Full article

Iron (Fe) is the fourth most abundant element on earth and represents an essential nutrient for life. As a fundamental mineral element for cell growth and development, iron is available for uptake as ferric ions, which are usually oxidized into complex oxyhydroxide polymers, insoluble under aerobic conditions. In these conditions, the bioavailability of iron is dramatically reduced. As a result, microorganisms face problems of iron acquisition, especially under low concentrations of this element. However, some microbes have evolved mechanisms for obtaining ferric irons from the extracellular medium or environment by forming small molecules often regarded as siderophores. Siderophores are high affinity iron-binding molecules produced by a repertoire of proteins found in the cytoplasm of cyanobacteria, bacteria, fungi, and plants. Common groups of siderophores include hydroxamates, catecholates, carboxylates, and hydroximates. The hydroxamate siderophores are commonly synthesized by fungi. L-ornithine is a biosynthetic precursor of siderophores, which is synthesized from multimodular large enzyme complexes through non-ribosomal peptide synthetases (NRPSs), while siderophore-Fe chelators cell wall mannoproteins (FIT1, FIT2, and FIT3) help the retention of siderophores. S. cerevisiae, for example, can express these proteins in two genetically separate systems (reductive and nonreductive) in the plasma membrane. These proteins can convert Fe (III) into Fe (II) by a ferrous-specific metalloreductase enzyme complex and flavin reductases (FREs). However, regulation of the siderophore through Fur Box protein on the DNA promoter region and its activation or repression depend primarily on the Fe availability in the external medium. Siderophores are essential due to their wide range of applications in biotechnology, medicine, bioremediation of heavy metal polluted environments, biocontrol of plant pathogens, and plant growth enhancement. Full article

Enzymatic catalysis is one of the main pillars of sustainability for industrial production. Enzyme application allows minimization of the use of toxic solvents and to valorize the agro-industrial residues through reuse. In addition, they are safe and energy efficient. Nonetheless, their use in biotechnological processes is still hindered by the cost, stability, and low rate of recycling and reuse. Among the many industrial enzymes, fungal laccases (LCs) are perfect candidates to serve as a biotechnological tool as they are outstanding, versatile catalytic oxidants, only requiring molecular oxygen to function. LCs are able to degrade phenolic components of lignin, allowing them to efficiently reuse the lignocellulosic biomass for the production of enzymes, bioactive compounds, or clean energy, while minimizing the use of chemicals. Therefore, this review aims to give an overview of fungal LC, a promising green and sustainable enzyme, its mechanism of action, advantages, disadvantages, and solutions for its use as a tool to reduce the environmental and economic impact of industrial processes with a particular insight on the reuse of agro-wastes. Full article