Adaptive laboratory evolution of Naematelia aurantialba under high temperature for efficient production of exopolysaccharide.

Liquid fermentation could revolutionize mushroom polysaccharide production, but the low temperature constraint hampers the process. This study implemented adaptive laboratory evolution (ALE) to enhance the thermotolerance of Naematelia aurantialba strains and increase expolysaccharide production. After 75 ALE cycles at 30 °C, the adaptive strain surpassed the wild-type strain by 5 °C. In a 7.5 L fermentor at 30 °C, the ALE strain yielded 17 % more exopolysaccharide than the wild type strain at 25 °C. Although the exopolysaccharide synthesized by both strains shares a consistent monosaccharide composition, infrared spectrum, and glycosidic bond composition, the ALE strain's exopolysaccharide has a larger molecular weight. Furthermore, the ALE strain's exopolysaccharide exhibits superior cryoprotection performance compared to that produced by the original strain. The adapted strain demonstrated lower ROS levels and increased activity of antioxidant enzymes, indicating improved performance. Fatty acid profiling and transcriptomics revealed reconfiguration of carbohydrate metabolism, amino acid metabolism, and membrane lipid synthesis in thermophilic strains, maintaining cellular homeostasis and productivity. This study provides efficient strains and fermentation methods for high-temperature mushroom polysaccharide production, reducing energy consumption and costs.

In vitro digestion and fecal fermentation of basidiospore-derived exopolysaccharides from Naematelia aurantialba.

Mushroom polysaccharides exhibit numerous health-enhancing attributes that are intricately linked to the breakdown, assimilation, and exploitation of polysaccharides within the organism. Naematelia aurantialba polysaccharides (NAPS-A), highly prized polysaccharides derived from mushrooms, remain shrouded in uncertainty regarding their characteristics pertaining to gastrointestinal digestion and gut microbial fermentation. The study aimed to understand the digestion and fecal fermentation patterns of NAPS-A. After simulated digestion, NAPS-A's physicochemical properties remained unchanged. However, during in vitro fecal fermentation, indigestible NAPS-A underwent significant changes in various properties, such as reducing sugar, chemical composition, constituent monosaccharides, Molecular weight, apparent viscosity, FT-IR spectra, and microscopic morphology. Notably, NAPS-A was effectively utilized by the gut microbiota, with unchanged properties after digestion but altered after fermentation. It influenced gut microbe composition by increasing beneficial bacteria (Lactobacillus, Faecalibacterium, and Roseburia), lowering pH, and producing short-chain fatty acids. NAPS-A fermentation enriches carbohydrate, fatty acid, and amino acid metabolic pathways through PICRUSt prediction analysis. Overall, these findings emphasize NAPS-A's role in regulating gut bacteria and their metabolic functions, despite its challenging digestibility.

Efficient production and skincare activity evaluation of schizophyllan, a ß-glucan derived from Schizophyllum commune NTU-1.

Schizophyllan (SPG), a ß-glucan produced by the fungus Schizophyllum commune, possesses a ß-(1 â†’ 3)-linked backbone with single ß-(1 â†’ 6)-linked glucose side chains at approximately every third residue. In this study, we screened SPG-producing strains of S. commune from different provinces in China. A candidate strain (NTU-1) with a high SPG yield was chosen, and the fermentation conditions were optimized. The optimal carbon and nitrogen sources were sucrose (40 g/L) and yeast extract (20 g/L), respectively. The optimal conditions for pH and temperature were 5.0 and 28 °C, respectively. Inclusion of 0.2 mg/L of 2,4-Dichlorophenoxyacetic acid in the medium further increased the SPG concentration. In a 5-L bioreactor, the fermentation cycle was reduced from the initial seven days to five days, and the concentration of SPG obtained was 21.3 g/L, which is the highest reported to date. In addition, we evaluated the bioactivity of the SPG prepared using strain NTU-1. The results showed that SPG had certain characteristics of anti-oxidation, anti-photoaging, and inhibition of melanin production, making it a promising reagent for skin care.

Analysis of Volatile Flavor Substances in the Enzymatic Hydrolysate of Lanmaoa asiatica Mushroom and Its Maillard Reaction Products Based on E-Nose and GC-IMS.

An electronic nose (E-Nose) and gas chromatography-ion mobility spectrometry (GC-IMS) were used to analyze the volatile flavor compounds (VFCs) of the enzymatic hydrolysate of Lanmaoa asiatica and its Maillard reaction products (MRPs). E-Nose sensors have strong response signals to sulfide, nitrogen oxides, alcohols, and aldehyde ketone, and the aroma profile was increased after the Maillard reaction (MR). According to GC-IMS, A total of 84 known compounds were identified. Aldehydes, ketones and alcohols are the main VFCs. After MR, the concentrations of some alcohols decreased, and the concentration of pyrazines and ketones increased. Principal component analysis (PCA) and similarity analysis showed that the enzymatic hydrolysate and MRPs were different and could be effectively distinguished. In conclusion, this study clarified the changes in VFCs before and after the MR. The results can provide a theoretical basis for the quality control and flavor changes during the processing of Lanmaoa asiatica and provide a new method for flavor analysis of edible mushrooms and their products.

Nutritional Function and Flavor Evaluation of a New Soybean Beverage Based on Naematelia aurantialba Fermentation.

The soy beverage is a healthy product rich in plant protein; however, its unpleasant flavor affects consumer acceptance. The aim of this study was to determine the feasibility of using Naematelia aurantialba as a strain for the preparation of fermented soybean beverages (FSB). Increases in Zeta potential, particle size, and viscosity make soy beverages more stable. We found that nutrient composition was increased by fermenting N. aurantialba, and the antioxidant activity of soybean beverages significantly increased after 5 days of fermentation. By reducing the content of beany substances such as hexanal and increasing the content of 1-octen-3-ol, the aroma of soybean beverages fermented by N. aurantialba changed from "beany, green, and fatty" to "mushroom and aromatic". The resulting FSB had reduced bitterness but considerably increased sourness while maintaining the fresh and sweet taste of unfermented soybean beverages (UFSB). This study not only provides a theoretical basis for the market promotion of FSB but also provides a reference for basidiomycetes-fermented beverages.

Whole Genome Sequencing and Annotation of Naematelia aurantialba (Basidiomycota, Edible-Medicinal Fungi).

Naematelia aurantialba is a rare edible fungus with both nutritional and medicinal values and especially rich in bioactive polysaccharides. However, due to the lack of genomic information, researches on the mining of active compounds, artificial breeding and cultivation, genetics, and molecular biology are limited. To facilitate the medicinal and food applications of N. aurantialba, we sequenced and analyzed the whole genome of N. aurantialba for the first time. The 21-Mb genome contained 15 contigs, and a total of 5860 protein-coding genes were predicted. The genome sequence shows that 296 genes are related to polysaccharide synthesis, including 15 genes related to nucleoside-activated sugar synthesis and 11 genes related to glucan synthesis. The genome also contains genes and gene clusters for the synthesis of other active substances, including terpenoids, unsaturated fatty acids, and bioactive proteins. In addition, it was also found that N. aurantialba was more closely related to Naematelia encephala than to Tremella fuciformis. In short, this study provides a reference for molecular cognition of N. aurantialba and related researches.

High-efficiency production of Tremella aurantialba polysaccharide through basidiospore fermentation.

This study aims to develop a spore fermentation method instead of fruiting body extraction for the production of Tremella aurantialba polysaccharide (TAPS). The purified spore strain Tremella aurantialba NX-20 was isolated for TAPS fermentation. The fermented TAPS and the extracted TAPS had the same monosaccharide composition but higher molecular weight. The conditions for TAPS fermentation by NX-20 were optimized, which were 25 °C, pH 7, 10% initial inoculum and 5 days. To further reduce the processing steps and costs of the medium, tofu wastewater (TW) was used to replace defatted soybean meal (DSM). In a 7.5 L fermentation tank, 40 g/L glucose with 10-fold diluted TW was used as the medium. For TAPS fermentation, a maximum yield of 15.02 ± 0.40 g/L was achieved. In conclusion, this study provides a feasible strategy for the efficient preparation of TAPS through spore fermentation.

Effect of galactose side-chain on the self-assembly of xyloglucan macromolecule.

As a common side-chain residue of polysaccharide, galactose plays a significant role in multiple aspects of the macromolecules. This study showed how degalactosylation induced drastic self-assembly transition of xyloglucan from spherical aggregates toward ribbon-like aggregates, and how it led to largely decreased water solubility and apparent viscosity within a short range of galactose removal ratio. To better understand this phenomenon, the size of the ellipsoid-like aggregated nanoparticles were carefully measured and compared, and it was found out that those nanoparticles which lost more galactose residues turned out to be more slender and tend to bind and stack closely in parallel, thereby forming huge ribbon-like aggregates. The galactose residue is considered as the hydrophilic group, and the decreased number of which caused a more hydrophobic behavior.

Production, structure, and bioactivity of polysaccharide isolated from Tremella fuciformis XY.

Tremella fuciformis polysaccharide (TFP) is an important bioactive substance in Tremella, that contributes to its use as medicine and food. In this study, a novel fungus Tremella fuciformis XY was isolated and introduced to produce macromolecule polysaccharides (TFPB). The fermentation conditions were optimized and results demonstrates that the initial pH, optimal temperature and liquid volume were 6.0, 26 °C and 80 mL in a 500 mL flask, respectively. The maximum yield of TFPB was 9.05 ± 0.05 g/L, which is 59.05% higher than the basic yield (5.69 ± 0.02 g/L). The TFPB, purified by DEAE column, had a molecular weight (Mw) of 1.14 × 103 kDa and consisted mainly of mannose, glucuronic acid, glucose, galactose, xylose, and rhamnose at a molar ratio of 3.5:1.2:2:1.6:1.4:3. The structure of TFPB was preliminarily investigated by methylation analysis, infrared spectroscopy, and nuclear magnetic resonance analysis. The main linkage types were identified as 1,4-xylp, 1,4-manp, 1-xylp, 1-manp, 1,4-glcp, and 1,3,4-galp. Moreover, the antioxidant assays showed that TFPB could scavenge reactive oxygen species and hydroxyl radicals.

Characterization of Fractional Polysaccharides from Gleditsia sinensis and Gleditsia microphylla Gums.

The seeds of Gleditsia sinensis and Gleditsia microphylla, widespread in China, are an important source of galactomannans. G. sinensis gum (GSG) and G. microphylla gum (GMG) were purified and precipitated using different concentrations of ethanol and isopropanol. The GSG and GMG, precipitated in different stages, presented different characteristics, including polymer recovery, mannose/galactose ratio, chemical composition, molecular weight, and morphological appearance. The galactomannan recovery of GSG and GMG in 33.3% ethanol was 81.7% and 82.5%, respectively, while that in 28.8% isopropanol was 81.3% and 82.9%, respectively. To achieve similar precipitation efficiency, the amount of isopropanol should be lower than that of ethanol because of the lower dielectric constant of isopropanol (20 vs. 25 for ethanol). The precipitation behavior of galactomannans in polar organic solvents was dependent on the molecular structures and properties of the solvent. A higher mannose/galactose ratio and a higher molecular weight was obtained in a lower concentration of alcohols.

Enhancement of Gleditsia sinensis gum rheological properties with pressure cell treatment in semi-solid state.

The apparent viscosity, molecular weight, and molecular weight distribution are important physical properties that determine the functional properties of galactomannan gum. Gleditsia sinensis gum (GSG) in semi-solid state was pressure cell treated over a range of temperature (30-110 °C) under nitrogen maintained at a pressure of 1.0-4.0 MPa. Physicochemical properties of GSG samples both before and after the pressure cell treatment were characterized. These include measurements of rheological properties by LVDV-III Ultra Rheometer, molecular weight and radius of gyration by light scattering, and changes in surface morphology by scanning electron microscopy. GSG had the highest apparent viscosity at a treatment temperature of 30 °C; further increase in temperature led to decrease in apparent viscosity. The apparent viscosity of GSG can be efficiently improved at room temperature and low pressure. The process of pressure cell treatment of GSG in semi-solid state could be industrialized for enhancement of rheological properties of galactomannan gum.

High glucose recovery from direct enzymatic hydrolysis of bisulfite-pretreatment on non-detoxified furfural residues.

This study reports four schemes to pretreat wet furfural residues (FRs) with sodium bisulfite for production of fermentable sugar. The results showed that non-detoxified FRs (pH 2-3) had great potential to lower the cost of bioconversion. The optimal process was that unwashed FRs were first pretreated with bisulfite, and the whole slurry was then directly used for enzymatic hydrolysis. A maximum glucose yield of 99.4% was achieved from substrates pretreated with 0.1 g NaHSO3/g dry substrate (DS), at a relatively low temperature of 100 °C for 3 h. Compared with raw material, enzymatic hydrolysis at a high-solid of 16.5% (w/w) specifically showed more excellent performance with bisulfite treated FRs. Direct bisulfite pretreatment improved the accessibility of substrates and the total glucose recovery. Lignosulfonate in the non-detoxified slurry decreased the non-productive adsorption of cellulase on the substrate, thus improving enzymatic hydrolysis.

Physico-chemical characterization of the temperature dependent hydration kinetics of Gleditsia sinensis gum.

The physico-chemical properties and hydration kinetics of Gleditsia sinensis gum were investigated to evaluate its temperature dependence. The increase of temperature resulted in improved solubility of G. sinensis gum, and the dissolved galactomannan showed decreased degree of galactose substitution (DSGal) and increased molecular weight (p<0.05, ANOVA). The solubility of G. sinensis galactomannan at 25, 45, 60 and 80 °C was 73.4, 83.8, 98.1, and 99.4%, respectively. Weibull and Logarithmic models were well fitted (R(2)>0.96), and the hydration index t0.8 at different temperatures varied in the range of 51-302 min. It was found that galactomannan with low DSGal and high molecular weight exhibited slow hydration rate and poor solubility.

Beneficial effects of wheat gluten hydrolysate to extend lifespan and induce stress resistance in nematode Caenorhabditis elegans.

Previous studies have showed that wheat gluten hydrolysate (WGH) has the anti-oxidative property. In the present study, we examined the possible safety property of WGH and the beneficial effects of WGH to extend lifespan and induce stress resistance using nematode Caenorhabditis elegans as the in vivo assay system. We found that WGH at concentrations of 0.1-1 mg/mL did not cause lethality, influence development, alter locomotion behavior and brood size, and induce significant intestinal autofluorescence and reactive oxygen species (ROS) production in young adults. Treatment with 0.1-1 mg/mL of WGH significantly extended lifespans of nematodes under the normal conditions. Moreover, WGH treatment significantly inhibited the induction of intestinal autofluorescence and suppressed the decrease in locomotion behavior during the aging process of nematodes. Furthermore, pre-treatment with 1 mg/mL of WGH significantly suppressed the adverse effects caused by heat-stress or oxidative stress on nematodes as indicated by the alterations of both lifespan and intestinal ROS production. Therefore, WGH treatment is relatively safe and has beneficial effects on nematodes under both the normal conditions and the stress conditions.

Enzymatic production and characterization of manno-oligosaccharides from Gleditsia sinensis galactomannan gum.

Enzymatic hydrolysis of Gleditsia sinensis gum was performed to produce manno-oligosaccharides having functional applications as dietary fiber and prebiotics. The optimum hydrolysis conditions, including enzyme loading, temperature and time, from response surface methodology were 8.1 U/g, 57.4 °C and 34.1 h, respectively. The yield of DP 1-5 oligosaccharides was 75.9% (29.1 g/L). The Michaelis-Menten kinetics and molecular weight distribution were determined. The obtained oligosaccharides were further separated by HPLC and SEC, and the galactose distribution of G. sinensis gum was elucidated. Results indicated that G. sinensis gum has potential to produce value-added oligosaccharides in food industries.