Function of Transcription Factors PoMYB12, PoMYB15, and PoMYB20 in Heat Stress and Growth of Pleurotus ostreatus.

MYB transcription factors (TFs) have been extensively studied in plant abiotic stress responses and growth and development. However, the role of MYB TFs in the heat stress response and growth and development of Pleurotus ostreatus remains unclear. To investigate the function of PoMYB12, PoMYB15, and PoMYB20 TFs in P. ostreatus, mutant strains of PoMYB12, PoMYB15, and PoMYB20 were generated using RNA interference (RNAi) and overexpression (OE) techniques. The results indicated that the mycelia of OE-PoMYB12, OE-PoMYB20, and RNAi-PoMYB15 mutant strains exhibited positive effects under heat stress at 32 °C, 36 °C, and 40 °C. Compared to wild-type strains, the OE-PoMYB12, OE-PoMYB20, and RNAi-PoMYB15 mutant strains promoted the growth and development of P. ostreatus. These mutant strains also facilitated the recovery of growth and development of P. ostreatus after 24 h of 36 °C heat stress. In conclusion, the expression of PoMYB12 and PoMYB20 supports the mycelium's response to heat stress and enhances the growth and development of P. ostreatus, whereas PoMYB15 produces the opposite effect.

Mnsod1 promotes the development of Pleurotus ostreatus and enhances the tolerance of mycelia to heat stress.

BACKGROUND: Superoxide dismutases (SODs, EC 1.15.1.1) are defense proteins that can be used as sweepers to clear reactive oxygen species (ROS). They have been widely studied in the plant. Intensive research demonstrates that SOD plays an essential role in plants. However, in Pleurotus ostreatus, the function and regulatory pathway of SOD in the growth and development and the abiotic stress response have not been clear. RESULTS: In this study, three MnSOD-encoding genes of the P. ostreatus CCMSSC00389 strain were cloned and identified. Mnsod1, Mnsod2, and Mnsod3 were interrupted by 3, 7, and 2 introns, and encoded proteins of 204, 220, and 344 amino acids, respectively. By comparing the relative expression of three MnSOD-encoding genes in mycelia, the results showed that the gene with the highest primary expression was Mnsod1. Subsequently, the function of P. ostreatus Mnsod1 was explored by overexpression (OE) and RNA interference (RNAi). The results showed that during the growth and development of P. ostreatus, MnSOD1 protein increased gradually from mycelia to the fruiting body, but decreased in spores. The change of Mnsod1 transcription level was not consistent with the changing trend of MnSOD1 protein. Further studies showed that during primordia formation, the expression of Mnsod1 gradually increased, reaching a peak at 48 h, and the transcription level was 2.05-folds compared to control. H2O2 content progressively accumulated during the formation of primordia, and its change trend was similar to that of Mnsod1 transcription. OE-Mnsod1-1 and OE-Mnsod1-21 strains accelerated the formation of primordia. The results suggested that Mnsod1 may participate in the formation rate of P. ostreatus primordium by regulating the signal molecule H2O2. In addition, OE-Mnsod1-1 and OE-Mnsod1-21 strains shortened the mycelial recovery time after heat stress and improved the tolerance of the strains to 2.5 mM and 5 mM H2O2, which showed that Mnsod1 was involved in the response of P. ostreatus mycelium to heat stress. CONCLUSIONS: This study indicates that Mnsod1 plays an active role in the formation of P. ostreatus primordia and the response to abiotic stress.

Transcription factor FfMYB15 regulates the expression of cellulase gene FfCEL6B during mycelial growth of Flammulina filiformis.

BACKGROUND: Cellulose degradation can determine mycelial growth rate and affect yield during the growth of Flammulina filiformis. The degradation of cellulose requires the joint action of a variety of cellulases, and some cellulase-related genes have been detected in mushrooms. However, little is known about the transcriptional regulatory mechanisms of cellulose degradation. RESULTS: In this study, FfMYB15 that may regulate the expression of cellulase gene FfCEL6B in F. filiformis was identified. RNA interference (RNAi) showed that FfCEL6B positively regulated mycelial growth. Gene expression analyses indicated that the expression patterns of FfCEL6B and FfMYB15 in mycelia cultured on the 0.9% cellulose medium for different times were similar with a correlation coefficient of 0.953. Subcellular localization and transcriptional activity analyses implied that FfMYB15 was located in the nucleus and was a transcriptional activator. Electrophoretic mobility shift assay (EMSA) and dual-luciferase assays demonstrated that FfMYB15 could bind and activate FfCEL6B promoter by recognizing MYB cis-acting element. CONCLUSIONS: This study indicated that FfCEL6B played an active role in mycelial growth of F. filiformis and was regulated by FfMYB15.

The Effects of a High-Fat/Cholesterol Diet on the Intestine of Rats Were Attenuated by Sparassis latifolia Polysaccharides.

Research background: Sparassis latifolia polysaccharides can regulate lipids and cholesterol in serum and liver. However, little is known about the regulation mechanism of the polysaccharides on cholesterol metabolism and especially the causal relationship with gut microbiota regulation. This study will provide a theoretical basis for the cholesterol-lowering mechanism of S. latifolia polysaccharides and further development of functional foods. Experimental approach: In this study, we investigated how the regulation mechanism of Sparassis latifolia polysaccharides affects intestinal cholesterol metabolism in high-fat and high-cholesterol diet-fed rats. Briefly, enzymatic colorimetric microplate assay was used to determine the concentration of faecal bile acid. Gas chromatography-mass spectrometry was used to detect the content of cholesterol and alcohol in faeces. Haematoxylin and eosin staining method was applied to observe the changes in the structure of the small intestine tissue. The related gene expressions in jejunum and ileum were detected by real-time fluorescent quantitative polymerase chain reaction. The related protein expressions in jejunum were studied by using Western blot. High-throughput sequencing was used to detect the intestinal flora changes of the caecal contents. Gas chromatography-mass spectrometry was applied to detect the concentration of short-chain fatty acids in the caecal content. Results and conclusions: The results showed that Sparassis latifolia polysaccharides could improve the intestinal morphological structure and physiological indices in rats fed high-fat and high-cholesterol diet. Moreover, it could improve intestinal cholesterol metabolism disorder induced by high-fat and high-cholesterol diets via the reduction of the expression of HMGCR, NPC1L1, ACAT2, MTP, ASBT and IBABP mRNA or protein, increasing ABCG8 mRNA expression. In addition, it could also increase the relative abundance of Bacteroides, Butyricicoccus, Parabacteroides, Parasutteerella and Alloprevotella and the short-chain fatty acid concentration, to comprehensively regulate the intestinal cholesterol metabolism. The metabolomics analysis found that Sparassis latifolia polysaccharides could affect lipid, carbohydrate and other related metabolites. Some biomarkers associated with cholesterol metabolism correlated significantly with the abundance of specific intestinal microbiota. Novelty and scientific contribution: These findings indicate that Sparassis latifolia polysaccharides could attenuate intestinal cholesterol metabolism disorder, correlating with modulating gut microbiota and improving host metabolism. They provide theoretical support for the development of Sparassis latifolia as a new food resource.

Comparison of physicochemical and biochemical properties of natural and arginine-modified melanin from medicinal mushroom Ganoderma lucidum.

Melanin is a hydrophobic biomolecule produced widely in fungi. Compared with other fungi, health benefits have been associated with medicinal mushrooms, which may provide an excellent source of natural melanin. Nevertheless, the hydrophobicity of melanin may limit its applications. Consequently, the present study was carried out on isolation of melanin from the medicinal mushroom Ganoderma lucidum (GLM) and modification with arginine to improve its solubility. The physicochemical and biochemical properties of melanin were evaluated including structural characterization, solubility, stability, antioxidant activities, and inhibitory effect on pancreatic lipase activity. Arginine-modified melanin showed better solubility, higher color value, stronger antioxidant activity, and stronger inhibitory effect on pancreatic lipase activity in vitro than GLM. In addition, both have good stability in the dark and natural light. These results opened possibilities for providing an excellent source of natural melanin in health food or food additives fields.

iTRAQ-Based Comparative Proteomics Analysis of the Fruiting Dikaryon and the Non-fruiting Monokaryon of Flammulina velutipes.

Flammulina velutipes is a potentially excellent fungus to study basic mechanisms of basidiomycete mycelium biology. To provide a better understanding of the mechanism of hyphae growth and fruit-body formation, the biological functions of the differentially abundant proteins between the fruiting dikaryon and the non-fruiting monokaryon of F. velutipes were investigated at the proteomic level using iTRAQ-coupled two-dimensional liquid chromatography tandem mass spectrometry technique. Among the 1198 proteins identified with high confidence, a total of 472 proteins were detected differentially abundant at least one of the mycelium development stages. In-depth data analysis revealed that differentially expressed proteins were influenced a variety of cellular processes, particularly metabolic processes. Functional pathway analysis indicated that 63 up-regulated proteins at only the fruiting dikaryon (Fv13) stage were mainly distributed in 51 specific Kyoto Encyclopedia of Genes and Genome pathways, such as amino acids biosynthesis and metabolism, signaling pathway, and central carbon metabolism. These up-regulated proteins could possibly serve as potential biomarkers to study the mycelium development pathways as well as provide new insights on the mycelium heterogenic compatibility and fruit-body formation mechanisms of basidiomycetes.

All-natural polysaccharide and protein complex nanoparticles from Clitocybe squamulosa as unique Pickering stabilizers for oil-in-water emulsions.

This study aimed to develop novel nanoparticles that can serve as an excellent oil-in-water (O/W) Pickering stabilizer. The polysaccharide-protein complex nanoparticles (PPCNs-20 and PPCNs-40) were prepared at different ultrasonication amplitudes (20 % and 40 %, respectively) from the polysaccharide-protein complexes (PPCs) which were extracted from the residue of Clitocybe squamulose. Compared with PPCs and PPCNs-20, the PPCNs-40 exhibited dispersed blade and rod shape, smaller average size, and larger zeta potential, which indicated significant potential in O/W Pickering emulsion stabilizers. Subsequently, PPCNs-40 stabilized Pickering emulsions were characterized at different concentrations, pHs, and oil phase contents. The average size, micromorphology, rheological properties, and storage stability of the emulsions were improved as the concentration of PPCNs-40, the ratio of the soybean oil phase and pH value increased. Pickering emulsions showed the best stability when the concentration of PPCNs-40 was 3 wt%, and the soybean oil fraction was 30 % under both neutral and alkaline conditions. The emulsions demonstrated shear thinning and gelation behavior. These findings have implications for the use of eco-friendly nanoparticles as stabilizers for Pickering emulsions and provide strategies for increasing the added value of C. squamulosa.

Functional Properties, Rheological Characteristics, Simulated Digestion, and Fermentation by Human Fecal Microbiota of Polysaccharide from Morchella importuna.

Morchella importuna polysaccharide (MIP) has been proven to have obvious hypoglycemic effects on mice with type 2 diabetes (T2DM). This study looked at the functional and rheological characteristics of MIP, and investigated the effects of MIP on the human fecal microbiota through in vitro fermentation experiments. The outcomes demonstrate the excellent oil-holding capacity, emulsifying, foaming, and rheological characteristics of MIP. After salivary gastrointestinal digestion, the Mw of MIP decreased from 398.2 kDa and 21.5 kDa to 21.9 kDa and 11.7 kDa. By 16S rRNA sequencing of bacteria fermented in vitro, it was found that MIP did not improve the richness and diversity of intestinal microorganisms, but it may exert an anti-T2DM function by significantly increasing the relative abundance of Firmicutes and promoting Ruminococcaceae_UCG_014, Bacteroides, and Blautia proliferation. Escherichia-Shigella could also be inhibited to improve the intestinal microenvironment. In addition, the fermentation of MIP increased the total short-chain fatty acid (SCFA) concentration from 3.23 mmol/L to 39.12 mmol/L, and the propionic acid content increased significantly. In summary, MIP has excellent processing performance and is expected to exert potential anti-T2DM activity through the human intestinal microbiota, which has broad market prospects.

The effect of sequential extraction on the physicochemical and rheological properties of Naematelia aurantialba polysaccharides.

To overcome the difficulty of separation and low rate of extraction caused by highly viscous polysaccharides from Naematelia aurantialba (NA), four N. aurantialba polysaccharides (NAPs) were sequentially extracted using water (enzyme-/ultrasound-assisted extraction), alkali (0.1 mol/L NaOH), and acid (0.1 mol/L HCl), and named E-NAP, U-NAP, Al-NAP, and Ac-NAP. The properties of four NAPs were different. The yields of NAPs were 26.05 % (Ac-NAP) > 20.33 % (Al-NAP) > 17.99 % (U-NAP) > 12.77 % (E-NAP), respectively. The monosaccharide composition of NAPs was composed primarily of mannose, xylose, glucose, glucuronic acid, and galactose. Sequential extraction improved the purity and solubility of NAPs, but decreased the particle size, thermal stability, water retention, and crystallinity. Two polysaccharides, U-NAP and Al-NAP, had a triple helix structure. All the NAPs were pseudoplastic fluids with concentration/frequency-dependent entangled structure. Al-NAP with the highest viscosity exhibited an elastic gel, while Ac-NAP with the lowest viscosity was a viscous gel. The behavior of NAPs differed from that predicted using the Cox-Merz rule, and in particular, E-NAP and U-NAP more significantly deviated from the rule. In this study, four NAPs with different properties were extracted sequentially, which provided a theoretical basis for the down-stream processing with high added-value and utilization of NA and NAP.

Polysaccharides from an edible mushroom, Hericium erinaceus, alleviate ulcerative colitis in mice by inhibiting the NLRP3 inflammasomes and reestablish intestinal homeostasis.

This study aimed to assess the effects of polysaccharides extracted from Hericium erinaceus fruiting bodies (HEFPs) on the inflammatory response to oxidative stress in a mouse model of ulcerative colitis (UC) induced by ingestion of dextran sodium sulfate. The results indicated reduced oxidative damage in the HEFPs groups, as evidenced by significantly decreased malondialdehyde levels and significantly increased levels of the antioxidant enzymes superoxide dismutase and catalase in colon homogenates, compared with those in the Model Control (MC) group. Additionally, compared with the levels in the MC group, the levels of the pro-inflammatory factors IL-6, IL-1ß, and TNF-α in the positive-control (PC) and HEFPs groups were significantly lower, and that of the anti-inflammatory factor IL-10 was significantly higher. qRT-PCR analyses revealed that the colon expression patterns of IL-6, IL-1ß, TNF-α, and IL-18 were consistent with the serum levels. Western-blotting results indicated significantly lower levels of NLRP3, ASC, and caspase 1 P20 in the HEFPs and PC groups than in the MC group. These findings suggest that HEFPs alleviate UC by suppressing the NLRP3 inflammasome/Caspase-1 pathway. Lachnospiraceae, Clostridiales, Parabacteroides, Oscillibacter, and Clostridium XlVa genera were more abundant in the gut microbiota of the HEFPs group than that of the MC group.

Polysaccharide from Clitocybe squamulosa: Gel-forming properties and its compound effect with food thickener.

To study the gel-forming properties of polysaccharide from the fruiting body of Clitocybe squamulosa (CSFP) and its degradation product (UH-CSFP), the changes in steady-state and dynamic rheological properties of CSFP and UH-CSFP under different conditions (polysaccharide mass fraction, temperature, pH, and salt ion concentration) were studied. Polysaccharides with good gel-forming properties were selected and mixed with common edible thickeners (gelatin, guar gum, and locust bean gum), after which the properties of the composite gel were assessed. The steady-state rheological results showed that CSFP and UH-CSFP were pseudoplastic fluids, their apparent viscosity decreased with increasing temperature, the viscosity was greatest when the pH was 7. The addition of Na+ and Ca2+ could increase the viscosity, and the viscosity of UH-CSFP was lower than that of CSFP at the same mass fraction. The results of dynamic rheology indicated that G´ and G´´ of CSFP and UH-CSFP increased with increasing mass fraction, pH, and ion concentration (0.01 M to 1 M), and G´´ was always smaller than G´ indicating weak gel behavior. The thixotropy-related experimental results showed that the thixotropy ring area of CSFP and UH-CSFP increased with increasing mass fraction, the ring area of CSFP was larger than that of UH-CSFP, and the gel strength of CSFP was greater than that of UH-CSFP. The results of CSFP and three types of edible gels showed that the composite gels were pseudoplastic fluids, and their apparent viscosity was ranked (in descending order) as follows: guar bean gum, locust bean gum, and gelatin. The addition of CSFP improved the gel-forming properties of guar gum but did not significantly improve the gel properties of locust bean gum and gelatin. This study provides a theoretical basis for the selection of processing methods and the application of polysaccharides.

Effect of Different Drying Methods on the Quality of Oudemansiella raphanipes.

In this study, we used fresh Oudemansiella raphanipes as raw materials and pre-treated through hot air drying (HD), infrared radiation drying (ID), and vacuum freeze drying (VD) to investigate the effects of different drying methods on the rehydration rate, appearance quality, microstructure, and volatile flavor components of the dried products, as well as to determine the physicochemical properties and bioactivities of the polysaccharides in the dried O. raphanipes. The results showed that the VD O. raphanipes had the highest rehydration rate and the least shrinkage in appearance, and it better maintained the original color of the gills, but their aroma was not as strong as that of the HD samples. The scanning electron microscopy results indicate that VD maintains a good porous structure in the tissue, while HD and ID exhibit varying degrees of shrinkage and collapse. Seventy-five common volatile substances were detected in the three dried samples, mainly alkanes, alcohols, and esters. The polysaccharides (PS-H, PS-I, and PS-V) extracted from the dried samples of these three species of O. raphanipes had similar infrared spectral features, indicating that their structures are basically consistent. The highest yield was obtained for PS-V, and the polysaccharide content and glucuronic acid content of PS-I were higher than those of the remaining two polysaccharides. In addition, PS-V also showed better antioxidant activity and inhibitory activity against α-glucosidase as well as α-amylase. In conclusion, among the above three drying methods, the quality of O. raphanipes obtained by vacuum freeze drying is the best, and this experiment provides a theoretical basis for the selection of drying methods for O. raphanipes.

Effects of different extraction methods on the physico-chemical characteristics and biological activities of polysaccharides from Clitocybe squamulosa.

This study comparatively evaluated the effects of the commonly used six extraction methods (acidic, alkaline, enzymatic, ultrasonic, high-pressure, and microwave) on the physico-chemical properties, processing characteristics, and biological activities of polysaccharides from Clitocybe squamulosa (CSFPs). The results show that polysaccharides extracted using an enzyme-assisted extraction method has a relatively high extraction yield (4.46 ± 1.62 %) and carbohydrate content (70.79 ± 6.25 %) compared with others. Furthermore, CSFPs were all composed of glucose, galactose, mannose, xylose, and glucosamine hydrochloride. Only ultrasonic-assisted extraction of polysaccharides (CSFP-U) has a triple helix chain conformation. Scanning electron microscopy (SEM) revealed significant differences in the microstructure of polysaccharides prepared using different methods. Besides that, the polysaccharides prepared by alkali extraction (CSFP-B) and high-pressure assisted extraction (CSFP-H) have good water (2.86 ± 0.29 g/g and 3.15 ± 0.29 g/g) and oil (8.13 ± 0.32 g/g and 7.97 ± 0.04 g/g) holding properties. The rheological behavior demonstrated that CSFPs solutions were typical non-Newtonian fluid. Apart from this, the antioxidant capacity (clearing DPPH (IC50 = 0.29) and ABTS free radicals (IC50 = 0.19), total reduction ability (IC50 = 3.02)) of polysaccharides prepared by the microwave-assisted extraction (CSFP-M) method was significantly higher than that of other extraction methods. By contrast, the polysaccharide prepared by acid extraction (CSFP-A) has the optimum binding capacity (bile acid salt (71.30 ± 6.78 %) and cholesterol (57.07 ± 3.26 mg/g)). The antibacterial activity of CSFPs was positively correlated with their concentration. Thus, the research results can provide a theoretical basis for the development and utilization of polysaccharides from C. squamulosa.

The Effects of Naematelia aurantialba on the Pasting and Rheological Properties of Starch and the Research and Development of Soft Candy.

To study the effects of Naematelia aurantialba (NA) on the rheological and gelatinization properties of starch, the processing methods of NA were diversified. In this study, the gelatinization and rheological properties of corn starch (CS) and edible cassava starch (ECS) were investigated by adding NA with different mass fractions. Starch soft candy was prepared using NA, CS, and ECS as the main raw materials. Rheological studies showed that both CS-NA and ECS-NA exhibited elastic modulus (G') > viscosity modulus (G″), implying elastic behavior. G' was such that CS+1%NA > CS+5%NA > CS+3%NA > CS > CS+2%NA > CS+4%NA > ECS+4%NA > ECS+3%NA > ECS+5%NA > ECS+2%NA > ECS+1%NA > ECS. The gelatinization implied showed that after adding NA, the pasting temperature of CS-NA and ECS-NA increased by 1.33 °C and decreased by 2.46 °C, while their breakdown values decreased by 442.35 cP and 866.98 cP, respectively. Through a single-factor test and orthogonal test, the best formula of starch soft candy was as follows: 0.4 f of NA, 10 g of white granulated sugar, a mass ratio of ECS to CS of 20:1 (g:g), 0.12 g of citric acid, 1 g of red date power, and 16 mL of water. The soft candy was stable when stored for two days. This study offers a new direction for the research and development of NA starch foods.

Transcriptome Analysis Revealed That Hydrogen Peroxide-Regulated Oxidative Phosphorylation Plays an Important Role in the Formation of Pleurotus ostreatus Cap Color.

Pleurotus ostreatus is widely cultivated in China. H2O2, as a signaling molecule, can regulate the formation of cap color, but its regulatory pathway is still unclear, severely inhibiting the breeding of dark-colored strains. In this study, 614 DEGs specifically regulated by H2O2 were identified by RNA-seq analysis. GO-enrichment analysis shows that DEGs can be significantly enriched in multiple pathways related to ATP synthesis, mainly including proton-transporting ATP synthesis complex, coupling factor F(o), ATP biosynthetic process, nucleoside triphosphate metabolic processes, ATP metabolic process, purine nucleoside triphosphate biosynthetic and metabolic processes, and purine ribonuclease triphosphate biosynthetic metabolic processes. Further KEGG analysis revealed that 23 DEGs were involved in cap color formation through the oxidative phosphorylation pathway. They were enriched in Complexes I, III, IV, and V in the respiratory chain. Further addition of exogenous uncoupling agents and ATP synthase inhibitors clarifies the important role of ATP synthesis in color formation. In summary, H2O2 may upregulate the expression of complex-encoding genes in the respiratory chain and promote ATP synthesis, thereby affecting the formation of cap color. The results of this study lay the foundation for the breeding of dark-colored strains of P. ostreatus and provide a basis for the color-formation mechanism of edible fungi.

Highly sensitive detection of fluoride based on poly(3-aminophenylboronic acid)-reduced graphene oxide multilayer modified electrode.

A novel electrochemical method for detecting fluoride was developed based on gold electrode modified by layer-by-layer (LBL) assembly of poly(3-aminophenylboronic acid)-reduced graphene oxide (PAPBA-RGO) multilayers. The PAPBA-RGO multilayer-modified gold electrode was constructed by using alternating LBL assembly of RGO and PAPBA on a bare gold electrode by one-step electrodeposition. Fluoride was electrochemically determined based on the proposed modified electrode by evaluating the changes in peak current for potassium ferricyanide reduction caused by the conjunction of fluoride and boronic acid groups of PAPBA. The results indicated that the peak current for potassium ferricyanide reduction obviously decreased with the increasing fluoride concentration. The response range of our method for fluoride was 1 × 10-8 to 1 × 10-1 M with a detection limit of 6 × 10-10 M and high sensitivity and selectivity. This method was applied to detect fluoride in tap water, rice, apple, and edible fungi samples.

The Tissue Browning and Concomitant Toughening of Yellow Flammulina filiformis Stipes Is Caused by Oxidative Damage-Mediated Metabolic Disorder and Cell Wall Glycan Remodeling.

The browning and associated toughening of fruiting body stipes are the main causes of declines in the commercial production of yellow Flammulina filiformis. The dynamic metabolic changes from the top to bottom stipe sections of yellow F. filiformis fruiting bodies were investigated by integrating physiological, transcriptomic, and metabolomic analyses. The results indicated that oxidative stress levels gradually increased accompanying the degree of tissue browning and toughening from the top to bottom sections of F. filiformis stipes. In-depth analysis showed that there were remarkable changes in the expression of genes, and the content of metabolites correlated with the primary and secondary metabolism of F. filiformis stipes. Interestingly, the expression levels of genes participating in chitosan biosynthesis and the degree of deacetylation of chitosan increased from top to bottom in F. filiformis stipes, implying that cell wall glycan remodeling may contribute to concomitant toughening of the browning of F. filiformis stipes.

Effects of Freeze-Thaw Cycles on the Structures and Functional Properties of Clitocybe squamulosa Protein Isolates.

Changes in the functional properties and structures of Clitocybe squamulosa protein isolate (CSPI) in the process of freeze-thaw (F-T) cycles were explored. Remarkable alterations and the reduced content of protein ordered structure were revealed through structural analysis of CSPI after F-T treatments. The surface hydrophobicity and free sulfhydryl content of CSPI first increased and then decreased. However, after the F-T treatments, the carbonyl content of CSPI continued to increase. Similarly, the water holding capacity (WHC), oil holding capacity (OHC), and solubility of CSPI all declined as the number of F-T cycles increased. The foaming properties and emulsifying properties of CSPI were significantly improved and reached maximum values after three F-T cycles. CSPI undergoing two F-T cycles showed the highest digestibility, maximum polypeptide content, and highest DPPH and ·OH-radical-scavenging activities. The ·OH-radical-scavenging activities and reducing power of the gastrointestinally digested CSPI had the highest value after one F-T cycle. Therefore, it has been demonstrated that F-T treatments could be a residue-free and cost-effective tool for improving mushroom protein functional properties.

MAC Family Transcription Factors Enhance the Tolerance of Mycelia to Heat Stress and Promote the Primordial Formation Rate of Pleurotus ostreatus.

Pleurotus ostreatus is a typical tetrapolar heterologous edible mushroom, and its growth and development regulatory mechanism has become a research hotspot in recent years. The MAC1 protein is a transcription factor that perceives copper and can regulate the expression of multiple genes, thereby affecting the growth and development of organisms. However, its function in edible mushrooms is still unknown. In this study, two transcription factor genes, PoMCA1a and PoMAC1b, were identified. Afterwards, PoMAC1 overexpression (OE) and RNA interference (RNAi) strains were constructed to further explore gene function. The results showed that the PoMAC1 mutation had no significant effect on the growth rate of mycelia. Further research has shown that OE-PoMAC1a strains and RNAi-PoMAC1b strains exhibit strong tolerance under 32 °C heat stress. However, under 40 °C heat stress, the OE of PoMAC1a and PoMAC1b promoted the recovery of mycelial growth after heat stress. Second, the OE of PoMAC1a can promote the rapid formation of primordia and shorten the cultivation cycle. In summary, this study indicated that there are functional differences between PoMAC1a and PoMAC1b under different heat stresses during the vegetative growth stage, and PoMAC1a has a positive regulatory effect on the formation of primordia during the reproductive growth stage.

Transglutaminase-Catalyzed Glycosylation Improved Physicochemical and Functional Properties of Lentinus edodes Protein Fraction.

Lentinula edodes has high nutritional value and abundant protein. In order to develop and utilize edible mushroom protein, this study was designed to investigate the effects of TGase-catalyzed glycosylation and cross-linking on the physicochemical and functional properties of Lentinus edodes protein fraction. The results showed that within a certain time, glycosylation and TGase-catalyzed glycosylation decreased the total sulfydryl, free sulfydryl, disulfide bond, surface hydrophobicity, ß-fold and α-helix, but increased the fluorescence intensity, random coil, ß-turn, particle size and thermal stability. The apparent viscosity and the shear stress of the protein with an increase in shear rate were increased, indicating that TGase-catalyzed glycosylation promoted the generation of cross-linked polymers. In addition, the TGase-catalyzed glycosylated proteins showed a compact texture structure similar to the glycosylated proteins at the beginning, indicating that they formed a stable three-dimensional network structure. The flaky structure of proteins became more and more obvious with time. Moreover, the solubility, emulsification, stability and oil-holding capacity of enzymatic glycosylated Lentinus edodes protein fraction were significantly improved because of the proper TGase effects of glycosylation grafting and cross-linking. These results showed that glycosylation and TGase-catalyzed glycosylation could improve the processing characteristics of the Lentinula edodes protein fraction to varying degrees.