Methionine inducing carbohydrate esterase secretion of Trichoderma harzianum enhances the accessibility of substrate glycosidic bonds.

BACKGROUND: The conversion of plant biomass into biochemicals is a promising way to alleviate energy shortage, which depends on efficient microbial saccharification and cellular metabolism. Trichoderma spp. have plentiful CAZymes systems that can utilize all-components of lignocellulose. Acetylation of polysaccharides causes nanostructure densification and hydrophobicity enhancement, which is an obstacle for glycoside hydrolases to hydrolyze glycosidic bonds. The improvement of deacetylation ability can effectively release the potential for polysaccharide degradation. RESULTS: Ammonium sulfate addition facilitated the deacetylation of xylan by inducing the up-regulation of multiple carbohydrate esterases (CE3/CE4/CE15/CE16) of Trichoderma harzianum. Mainly, the pathway of ammonium-sulfate's cellular assimilates inducing up-regulation of the deacetylase gene (Thce3) was revealed. The intracellular metabolite changes were revealed through metabonomic analysis. Whole genome bisulfite sequencing identified a novel differentially methylated region (DMR) that existed in the ThgsfR2 promoter, and the DMR was closely related to lignocellulolytic response. ThGsfR2 was identified as a negative regulatory factor of Thce3, and methylation in ThgsfR2 promoter released the expression of Thce3. The up-regulation of CEs facilitated the substrate deacetylation. CONCLUSION: Ammonium sulfate increased the polysaccharide deacetylation capacity by inducing the up-regulation of multiple carbohydrate esterases of T. harzianum, which removed the spatial barrier of the glycosidic bond and improved hydrophilicity, and ultimately increased the accessibility of glycosidic bond to glycoside hydrolases.

Orthogonal Reversed-Phase C18 and Pentafluorophenyl HPLC Separation for Phytochemical Profiling of Serrulatanes in Eremophila denticulata.

Serrulatanes constitute a class of unique diterpenoids derived from all-Z nerylneryl diphosphate rather than the conventional all-E diterpenoid precursor geranylgeranyl diphosphate and thus provide an intriguing expansion of the chemical space of plant specialized metabolites. Plants of the Australian Eremophila genus are rich sources of structurally diverse serrulatanes. Here, we report the identification of 15 hitherto undescribed serrulatanes (eremoculatanes A-N), together with 16 previously reported compounds, from the EtOAc extract of Eremophila denticulata leaves. Isolation was performed by a combined use of systematic HPLC-PDA-HRMS-based phytochemical profiling and orthogonal reversed-phase C18 and pentafluorophenyl separations. Among the new compounds isolated, eremoculatane A contains a C12 backbone, for which the configuration was established by comparison of experimentally measured and theoretically calculated ECD spectra. The antihyperglycemic and antibacterial activities of the E. denticulata extract were investigated by high-resolution inhibition profiling, and they indicated that major constituents, mainly serrulatanes and flavonoids, contributed to the observed activity of the extract. One flavonoid, eupafolin (4), displayed moderate α-glucosidase inhibitory activity with an IC50 value of 41.3 µM, and four serrulatanes (8, 9, 19g, and 19j) showed more than 50% PTP1B inhibition at 200 µM.

Structure Elucidation of Prenyl- and Geranyl-Substituted Coumarins in Gerbera piloselloides by NMR Spectroscopy, Electronic Circular Dichroism Calculations, and Single Crystal X-ray Crystallography.

Crude ethyl acetate extract of Gerbera piloselloides (L.) Cass. was investigated by dual high-resolution PTP1B/α-glucosidase inhibition profiling and LC-PDA-HRMS. This indicated the presence of a series of unprecedented prenyl- and geranyl-substituted coumarin derivatives correlated with both α-glucosidase and PTP1B inhibitory activity. Repeated chromatographic separation targeting these compounds led to the isolation of 13 new compounds, of which ten could be isolated as both enantiomers after chiral separation. The structures of all isolated compounds were characterized by HRMS and extensive 1D and 2D NMR analysis. The absolute configurations of the isolated compounds were determined by comparison of experimental and calculated electronic circular dichroism spectra. Compound 6 features a rare furan-oxepane 5/7 ring system, possibly formed through addition of a geranyl unit to C-3 of 5-methylcoumarin, representing a new type of geranyl-substituted coumarin skeleton. Compounds 19 and 24 are the first examples of dimeric natural products consisting of both coumarin and chromone moieties.

Solution-Processable Conductive Composite Hydrogels with Multiple Synergetic Networks toward Wearable Pressure/Strain Sensors.

A biocompatible, flexible, yet robust conductive composite hydrogel (CCH) for wearable pressure/strain sensors has been achieved by an all-solution-based approach. The CCH is rationally constructed by in situ polymerization of aniline (An) monomers in the polyvinyl alcohol (PVA) matrix, followed by the cross-linking of PVA with glutaraldehyde (GA) as the cross-linker. The unique multiple synergetic networks in the CCH including strong chemical covalent bonds and abundance of weak physical cross-links, i.e., hydrogen bondings and electrostatic interactions, impart excellent mechanical strength (a fracture tensile strength of 1200 kPa), superior compressibility (ε = 80%@400 kPa), outstanding stretchability (a fracture strain of 670%), high sensitivity (0.62 kPa-1 at a pressure range of 0-1.0 kPa for pressure sensing and a gauge factor of 3.4 at a strain range of 0-300% for strain sensing, respectively), and prominent fatigue resistance (1500 cycling). As the flexible wearable sensor, the CCH is able to monitor different types of human motion and diagnostically distinguish speaking. As a proof of concept, a sensing device has been designed for the real-time detection of 2D distribution of weight or pressure, suggesting its promising potentials for electronic skin, human-machine interaction, and soft robot applications.

Integration of homogeneous and heterogeneous advanced oxidation processes: Confined iron dancing with cyclodextrin polymer.

A novel catalyst which integrates heterogeneous and homogenous Fenton reactions is designed and fabricated by encapsulating 2,5-dihydroxy-1,4-benzoquinone (2,5-DBQ) in ECDP-Fe3O4, a composite of Fe3O4 nanoparticles immobilized on a ß-cyclodextrin polymer (ECDP) with ethylene diamine tetraacetic acid (EDTA) as cross-linking agent. The 2,5-DBQ@ECDP-Fe3O4 has superior catalytic performance for 4-nitrophenol and 2,4-dichlorophenol degradation compared with control systems. Mechanism study revealed that although the initial active site is Fe3O4 loaded on ECDP, the actually catalyst is the iron ions released from Fe3O4 but confined within the composite. EDTA in ß-cyclodextrin polymer can improve both the solubility and adsorption capacity to H2O2 of Fe3O4. The quinone molecules 2,5-DBQ in the ß-cyclodextrin cavity can accelerate Fe3+/Fe2+ cycle adjacent to the cavity, thus in favor of the decomposition of H2O2 into OH as main reactive oxidizing species. The current catalyst integrates the advantages of homogeneous and heterogeneous advanced oxidation processes and is promising in practical applications.

The functioning of a novel protein, Swollenin, in promoting the lignocellulose degradation capacity of Trichoderma guizhouense NJAU4742 from a proteomic perspective.

The functioning of a novel auxiliary enzyme, TgSWO from Trichoderma guizhouense NJAU4742, was investigated based on the proteomic analysis of wild-type (WT), knockout (KO) and overexpression (OE) treatments. The results showed that the cellulase and hemicellulase activities of OE and WT were significantly higher than those of KO. Simultaneously, tandem mass tag (TMT) analysis results indicated that cellulases and hemicellulases were significantly upregulated in OE, especially hydrophobin (HFB, A1A105805.1) and endo-ß-1,4-glucanases (A1A101831.1), with ratios of 43.73 and 9.88, respectively, compared with WT. The synergistic effect of TgSWO on cellulases increased the reducing sugar content by 1.45 times in KO + TgSWO (1.8 mg) compared with KO, and there was no significant difference between KO + TgSWO (1.2 mg) and WT. This study elucidated the function of TgSWO in promoting the lignocellulose degradation capacity of NAJU4742, which provides new insights into the efficient conversion of lignocellulose.