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Synthesis mechanism of antifungal active protein in the fermentation of agar waste liquid by Streptomyces rubiginosoheloolus

ABSTRACT: This project investigates the proteomic basis of antifungal activity generated by fermenting agar waste liquid with Streptomyces rubiginosoheloolus. Agar waste liquid at different concentrations was fermented, and extracellular proteins from active cultures were enriched by ammonium sulfate precipitation and anion-exchange chromatography, followed by mass-spectrometry-based identification. The ProteomeXchange dataset contains the raw and processed MS data for these antifungal protein fractions, which enabled the discovery of key proteins (histone-like nucleoid-structuring protein H-NS, a hypothetical protein and a ribosomal protein) that disrupt Alternaria spp. via membrane damage, ROS accumulation and lipid peroxidation, providing a basis for agar waste valorization into antifungal bioproducts.

ORGANISM(S): Streptomyces Rubiginosohelvolus

SUBMITTER: Zhipeng Li

PROVIDER: PXD071170 | iProX | Tue Nov 25 00:00:00 GMT 2025

REPOSITORIES: iProX

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Synthesis mechanism of antifungal active protein in the fermentation of agar waste liquid by Streptomyces rubiginosoheloolus.

Du Xiping X Lin Qiaoyan Q Qin Yonghong Y Li Zhipeng Z Yang Yuanfan Y Jiang Zedong Z Ni Hui H Li Qingbiao Q

Food chemistry 20251216

Phytopathogenic fungi represent a serious threat to global crop production, food security, and public health. Agar waste liquid as a biomass-rich byproduct, shows great potential for bioconversion into value-added antifungal compounds through microbial fermentation processes. This study demonstrated that fermentation of agar waste liquid by Streptomyces rubiginosoheloolus significantly enhanced antifungal activity, with the 50 % waste liquid group exhibited inhibition rates of 59.7 % and 57.1 % ...[more]

PMID: 41421068

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Project description:Invasive fungal infections threaten the lives and health of humans, especially immunodeficient patients or hospitalized patients with serious underlying diseases, and impose a heavy economic burden on society. The emergence of drug-resistant fungi, the formation of biofilms, and the limits and side effects of existing antifungal drugs increase the difficulty of clinical treatment, and there is an urgent need for the development of novel antifungal drugs. Therefore, based on previous kinase chemical library antifungal activity screening studies, this paper further investigates the activity of the ALK inhibitor HG-14-10-04 (HG) against various fungi and elucidates its mechanism of action. The in vitro antifungal activity of HG was evaluated by micro liquid-dilution method, time-killing curve, mycelium and biofilm formation. The results showed that HG exhibited inhibitory and even fungicidal effects against sensitive and resistant Candida albicans, Candida krusei, Cryptococcus neoformans, Candida tropicalis, Candida glabrata and Candida parapsilosis (MICs=8-16 μg/mL); HG significantly inhibited the mycelium and biofilm formation, and destroyed the mature biofilm; and it exhibited synergistic antifungal effects with amphotericin B. The antifungal mechanism of HG was investigated by flow cytometry and transmission electron microscopy, etc. Sequencing analysis showed a total of 1041 differentially expressed genes, of which 666 were up-regulated and 375 were down-regulated. According to the GO functional classification results, the up-regulated genes were mainly involved in ribosome production, oxidation-reduction and other functions, while the down-regulated genes were mainly involved in the synthesis of carbohydrate, glycoproteins, glycolipids and their metabolism, GPI anchor synthesis, and cytoskeleton and other functions. In addition, HG could significantly increase the level of ROS, induce the fungal necrosis, block the cell cycle at the G0/G1 phase, and change the ultrastructure of the fungi, especially the structure of the fungal cell wall. Therefore, the enhanced inhibitory and fungicidal activity of HG may be attributed to the elevation of ROS, alteration of cellular ultrastructure (especially cell wall structure), cell cycle arrest at the G0/G1 phase, and induction of fungal necrosis, providing a basis for the discovery of novel antifungal drugs or lead compounds.

Project description:Transcriptomics analysis of biopolymer (medium chain length polyhydroxyalkanoate) producing strain P.putida LS46 cultured with biodiesel derived waste carbon sources: studies of cellular adaptation to the industrial waste streams and metabolic profiling under the polymer producing conditions. We are reporting RNAseq analysis data here as part of our multi-level Omics study of medium chain length polyhydroxyalkanoate (mcl-PHA) producing strain P.putida LS46 culture with biodiesel derived waste glycerol and waste fatty acids. The data presented here will be used in two separate manuscripts. The objectives of this study are a): to evaluate cellular responses of P.putida LS46 under industrial waste stream. b): to study gene expression profile under two selected mcl-PHA producing conditions of P.putida LS46. Comparative multi-level Omics study: for objective a): Exponential P.putida LS46 cell from waste glycerol culture compared against reagent grade pure glycerol culture. For objective b): Two mcl-PHA producing conditions, namely stationary phase waste glycerol culture and exponential phase waste fatty acid culture of P.putida LS46, were compared against exponential phase waste glycerol culture of P.putida LS46. Major results from objective a): The waste glycerol substrate induced expression of a large number of genes putatively involved in heavy metal tolerance, including three gene clusters: a putative cusABC transcript unit and two copies of copAB, which are usually involved in copper resistance and tolerance to other monovalent heavy metals. A local gene relocation was observed in cluster 1 consisting cusABC and copAB relative to the KT2440 type strain according to the phylogenetic and gene neighbourhood analyses on various P. putida strains. P. putida LS46 also contains 11 putative MerR family regulators, which sense various environmental stimuli including heavy metals. MerR-1 is an ortholog of the copper response regulator of other gram-negative bacteria, and was highly up-regulated in waste glycerol cultures. Finally, a number of genes involved in cell responses to high extra-cellular Na+ concentrations, and genes of the fatty acid beta-oxidation pathway were up-regulated in waste glycerol cultures Major results from objective b): Regardless to the type of substrates, up-regulation of two mcl-PHA synthase (PhaC1 and PhaC2), and two phasin proteins (PhaF and PhaI) are the most common genotype under mcl-PHA production conditions. PhaG and possible PhaJ4 connect fatty acid de novo synthesis to mcl-PHA in waste glycerol culture. Interestingly, expression of gene, fabZ, in production of unsaturated fatty acid from fatty acid de novo synthesis was only observed in waste glycerol culture. On the other hand, PhaJ1 and PhaJ4 derived mcl-PHA production via fatty acid beta-oxidation was observed under waste fatty acid culture. These results would help to explain observed different production kinetics and monomer distribution of the polymer. Although under active mcl-PHA production condition, depression on the expression of glpF genes in glycerol transportation system prevent further channelling extra-cellular glycerol into the cell. Waste glycerol culture also triggers trahalose synthesis pathway, a potential competing pathway during mcl-PHA synthesizing. In waste fatty acid culture, the intermediates (acyl-CoA and 3-hydroxyacyl-CoA) of fatty acid beta-oxidation were used for mcl-PHA production and were also likely hydrolysed to their free acid forms via an up-regulated thioesteras coding gene, tesA. Acetyl-CoA cleaved from the pathway was clearly channeled into glyoxylate shut for C2 carbon assimilation over spillage as CO2 through TCA cycle or used in fatty acid biosynthesis pathway. In total 4 sampling points, namely exponential phase of pure glycerol, waste glycerol and waste free fatty acids cultures, and stationary phase of waste glycerol culture. For each sampling point, 2 biological replicates were taken. (Thus 8 samples in total)

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Synthesis mechanism of antifungal active protein in the fermentation of agar waste liquid by Streptomyces rubiginosoheloolus

Publications

Synthesis mechanism of antifungal active protein in the fermentation of agar waste liquid by Streptomyces rubiginosoheloolus.

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