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Intracellular Metabolomics Switching Alters Extracellular Acid Production and Insoluble Phosphate Solubilization Behavior in Penicillium oxalicum.

ABSTRACT: This research aims to understand the precise intracellular metabolic processes of how microbes solubilize insoluble phosphorus (Insol-P) to increase bio-available P. Newly isolated Penicillium oxalicum PSF-4 exhibited outstanding tricalcium phosphate (TP) and iron phosphate (IP) solubilization performance-as manifested by microbial growth and the secretion of low-molecular-weight organic acids (LMWOAs). Untargeted metabolomics approach was employed to assess the metabolic alterations of 73 intracellular metabolites induced by TP and IP compared with soluble KH2PO4 in P. oxalicum. Based on the changes of intracellular metabolites, it was concluded that (i) the enhanced intracellular glyoxylate and carbohydrate metabolisms increased the extracellular LMWOAs production; (ii) the exposure of Insol-P poses potential effects to P. oxalicum in destructing essential cellular functions, affecting microbial growth, and disrupting amino acid, lipid, and nucleotide metabolisms; and (iii) the intracellular amino acid utilization played a significant role to stimulate microbial growth and the extracellular LMWOAs biosynthesis.

SUBMITTER: Jiang Y

PROVIDER: S-EPMC7692655 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Project description:Cellulolytic enzyme hydrolysis of lignocellulose biomass to release fermentable sugars is one of the key steps in biofuel refining. Gene expression of fungal cellulolytic enzymes is tightly controlled at the transcriptional level. Key transcription factors such as activator ClrB/CLR2 and XlnR/XYR1, as well as repressor CreA/CRE1 play crucial roles in this process. The putative protein methyltransferase LaeA/LAE1 has also been reported to regulate the gene expression of the cellulolytic enzyme. The formation and gene expression of the cellulolytic enzyme was compared among Penicillium oxalicum wild type (WT) and seven mutants, including ?laeA (deletion of laeA), OEclrB (clrB overexpression), OEclrB?laeA (clrB overexpression with deletion of laeA), OExlnR (xlnR overexpression), OExlnR?laeA (xlnR overexpression with deletion of laeA), ?creA (deletion of creA), and ?creA?laeA (double deletion of creA and laeA). Results revealed that LaeA extensively affected the expression of glycoside hydrolase genes. The expression of genes that encoded the top 10 glycoside hydrolases assayed in secretome was remarkably downregulated especially in later phases of prolonged batch cultures by the deletion of laeA. Cellulase synthesis of four mutants ?laeA, OEclrB?laeA, OExlnR?laeA, and ?creA?laeA was repressed remarkably compared with their parent strains WT, OEclrB, OExlnR, and ?creA, respectively. The overexpression of clrB or xlnR could not rescue the impairment of cellulolytic enzyme gene expression and cellulase synthesis when LaeA was absent, suggesting that LaeA was necessary for the expression of cellulolytic enzyme gene activated by ClrB or XlnR. In contrast to LaeA positive roles in regulating prominent cellulase and hemicellulase, the extracellular ?-xylosidase formation was negatively regulated by LaeA. The extracellular ?-xylosidase activities improved over 5-fold in the OExlnR?laeA mutant compared with that of WT, and the expression of prominent ?-xylosidase gene xyl3A was activated remarkably. The cumulative effect of LaeA and transcription factor XlnR has potential applications in the production of more ?-xylosidase.

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Intracellular Metabolomics Switching Alters Extracellular Acid Production and Insoluble Phosphate Solubilization Behavior in Penicillium oxalicum.

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