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.

Necroptotic virotherapy of oncolytic alphavirus M1 cooperated with Doxorubicin displays promising therapeutic efficacy in TNBC.

Triple-negative breast cancer (TNBC) is the most aggressive molecular subtype among breast tumors and remains a challenge even for the most current therapeutic regimes. Here, we demonstrate that oncolytic alphavirus M1 effectively kills both TNBC and non-TNBC. ER-stress and apoptosis pathways are responsible for the cell death in non-TNBC as reported in other cancer types, yet the cell death in TNBC does not depend on these pathways. Transcriptomic analysis reveals that the M1 virus activates necroptosis in TNBC, which can be pharmacologically blocked by necroptosis inhibitors. By screening a library of clinically available compounds commonly used for breast cancer treatment, we find that Doxorubicin enhances the oncolytic effect of the M1 virus by up to 100-fold specifically in TNBC in vitro, and significantly stalls the tumor growth of TNBC in vivo, through promoting intratumoral virus replication and further triggering apoptosis in addition to necroptosis. These findings reveal a novel antitumor mechanism and a new combination regimen of the M1 oncolytic virus in TNBC, and highlight a need to bridge molecular diagnosis with virotherapy.

Carbothermal synthesis of biochar-supported metallic silver for enhanced photocatalytic removal of methylene blue and antimicrobial efficacy.

Pinewood biochar (PBC)-supported metallic silver (Ago) was prepared via a one-step carbothermal reduction route (AgH) or a wet-chemistry reduction method (AgW). XRD and SEM confirmed Ago was soldered on PBC matrix. Low methylene blue (MB) sorption was observed for unsupported Ago nanoparticles (AgNP), AgH and AgW. Under ultraviolet (UV) light irradiation, net MB degradation by AgH (15.88 g kg-1) was higher than that of AgW (12.50 g kg-1) and AgNP (10.27 g kg-1). TOC removal percentages after degradation corresponded largely to reduction of MB concentrations in solution, indicating MB was dominantly mineralized. Electron paramagnetic resonance (EPR) revealed that MB was degraded by reactive oxygen species (ROS) such as hydroxyl radical (OH), superoxide radical (O2-) and singlet oxygen (1O2). The scavenging experiments further suggested that OH scavengers suppressed MB degradation to a greater extent than other quenchers. Compared to AgW, AgH possessed greater abundance of persistent free radicals, which enhance ROS generation. PBC could also improve separation of electron-hole (e--h+) pairs and enhance electron transfer ascribing to favorable carbon structure. Besides, PBC-Ago maintained good antimicrobial efficacy over E.coli DH5α. This work presented a facile carbothermal route to prepare Ago-based photocatalysts for dye removal and microbial inhibition in industrial wastewater.