Siderophore-synthesizing NRPS reprogram lipid metabolic profiles for phenotype and function changes of Arthrobotrys oligospora.

The objective of this paper is to explore the function of the AOL-s00215g415 (Aog415) gene, which encodes for the synthesis of siderophore in the nematode trapping fungal model strain A. oligospora, in order to understand the relationship between siderophore biosynthesis and nematode trapping activity. After a through sequence analysis, it was determined that Aog415 is a siderophore-synthesizing NRPS. The product of this gene was then identified to be the hydroxamate siderophore desferriferrichrome, using mass spectrometry analysis. When compared to the WT strains, the Aog415 knockout strain exhibited a 60% decrease in siderophore content in fermentation broth. Additionally, the number of predatory rings of decreased by 23.21%, while the spore yield increased by 37.34%. The deletion of Aog415 did not affect the growth of A. oligospora in diverse nutrient medium. Lipid metabolism-related pathways were the primary targets of Aog415 disruption as revealed by the metabolomic analysis. In comparison to the WT, a significant reduction in the levels of glycerophospholipids, and glycolipids was observed in the mutation. The metabolic alteration in fatty acyls and amino acid-like molecules were significantly disrupted. The knockout of Aog415 impaired the biosynthesis of the hydroxamate siderophore desferriferrichrome, remodeled the flow of fatty acid in A. oligospora, and mainly reprogrammed the membrane lipid metabolism in cells. Desferriferrichrome, a hydroxamate siderophore affects the growth, metabolism and nematode trapping ability of A. oligospora by regulating iron intake and cell membrane homeostasis. Our study uncovered the significant contribution of siderophores to the growth and nematode trapping ability and constructed the relationship among siderophores biosynthesis, lipid metabolism and nematode trapping activity of A. oligospora, which provides a new insight for the development of nematode biocontrol agents based on nematode trapping fungi.

Latilactobacillus curvatus FFZZH5L isolated from pickled cowpea enhanced antioxidant activity in Caenorhabditis elegans by upregulating the level of glutathione S-transferase.

Latilactobacillus curvatus is a potential probiotic that possesses beneficial health properties and fermentation traits; however, the extent of understanding of the antioxidant activities of L. curvatus is limited. This study investigates the antioxidant activities of a new L. curvatus FFZZH5L strain. The strain exhibits broad tolerance to acids, bases and salts and demonstrated good adaption to the gastrointestinal environment, with a survival rate of 45% after 24 h of treatment in artificial gastrointestinal juice. Moreover, L. curvatus FFZZH5L exhibits inhibitory effects on Staphylococcus aureus, with a self-aggregation rate of 34.8% and a co-aggregation rate of 82.2%. In vitro, the DPPH radical scavenging ability and GSH-px enzyme activity of L. curvatus FFZZH5L reach 64.27% and 15.95 U mL-1, respectively. Treatment of C. elegans with L. curvatus FFZZH5L in vivo significantly extended the organism's lifespan. Furthermore, the activity of SOD, GSH-px and T-AOC was increased by 33.6%, 43.4% and 58.3%, respectively. Feeding C. elegans with L. curvatus FFZZH5L decreased the MDA, lipofuscin and ROS levels by 9%-36.4%. L. curvatus FFZZH5L effectively protected C. elegans against juglone-induced oxidative stress damage and led to a significant increase in the organism's survival under heat stress. The RT-qPCR analysis suggests that feeding C. elegans with L. curvatus FFZZH5L upregulates the expression levels of antioxidant-related genes including glutathione S-transferase 4 (gst-4), gst-1, gst-10, sod-3, sod-5, and sod-10 in C. elegans. Our investigation confirms the probiotic and antioxidant properties of L. curvatus, indicating its potential application in functional foods and the pharmaceutical industry.

Roles of the Fungal-Specific Lysine Biosynthetic Pathway in the Nematode-Trapping Fungus Arthrobotrys oligospora Identified through Metabolomics Analyses.

In higher fungi, lysine is biosynthesized via the α-aminoadipate (AAA) pathway, which differs from plants, bacteria, and lower fungi. The differences offer a unique opportunity to develop a molecular regulatory strategy for the biological control of plant parasitic nematodes, based on nematode-trapping fungi. In this study, in the nematode-trapping fungus model Arthrobotrys oligospora, we characterized the core gene in the AAA pathway, encoding α-aminoadipate reductase (Aoaar), via sequence analyses and through comparing the growth, and biochemical and global metabolic profiles of the wild-type and Aoaar knockout strains. Aoaar not only has α-aminoadipic acid reductase activity, which serves fungal L-lysine biosynthesis, but it also is a core gene of the non-ribosomal peptides biosynthetic gene cluster. Compared with WT, the growth rate, conidial production, number of predation rings formed, and nematode feeding rate of the ΔAoaar strain were decreased by 40-60%, 36%, 32%, and 52%, respectively. Amino acid metabolism, the biosynthesis of peptides and analogues, phenylpropanoid and polyketide biosynthesis, and lipid metabolism and carbon metabolism were metabolically reprogrammed in the ΔAoaar strains. The disruption of Aoaar perturbed the biosynthesis of intermediates in the lysine metabolism pathway, then reprogrammed amino acid and amino acid-related secondary metabolism, and finally, it impeded the growth and nematocidal ability of A. oligospora. This study provides an important reference for uncovering the role of amino acid-related primary and secondary metabolism in nematode capture by nematode-trapping fungi, and confirms the feasibility of Aoarr as a molecular target to regulate nematode-trapping fungi to biocontrol nematodes.