Metabolomics profiling reveals the mechanism of increased pneumocandin B0 production by comparing mutant and parent strains
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Metabolic profiling was used to discover mechanisms of increased pneumocandin B0 production in a high-yield strain by comparing it with its parent strain. Initially, 79 intracellular metabolites were identified, and the levels of 15 metabolites involved in six pathways were found to be directly correlated with pneumocandin B0 biosynthesis. Then by combining the analysis of key enzymes, acetyl-CoA and NADPH were identified as the main factors limiting pneumocandin B0 biosynthesis. Other metabolites, such as pyruvate, α-ketoglutaric acid, lactate, unsaturated fatty acids and previously unreported metabolite γ-aminobutyric acid were shown to play important roles in pneumocandin B0 biosynthesis and cell growth. Finally, the overall metabolic mechanism hypothesis was formulated and a rational feeding strategy was implemented that increased the pneumocandin B0 yield from 1821 to 2768 mg/L. These results provide practical and theoretical guidance for strain selection, medium optimization, and genetic engineering for pneumocandin B0 production.
KeywordsGlarea lozoyensis Pneumocandin B0 Metabolomics profiling Rational feeding strategies Enzyme activity
Nonribosomal peptide synthase
Pentose phosphate pathway
This work was supported by the National Science Foundation of China (No. 21776136), the National High Technology Research and Development Program (No. 2015AA021003), the Program for Innovative Research Team in University of Jiangsu Province (2015), the Natural Science Fund for Colleges and Universities in Jiangsu Province (No. 17KJB530006), the Postgraduate Research and Practice Innovation Program of Jiangsu Province (No. KYCX170960), the Natural Science Foundation of Jiangsu Province (BK20161048) and the Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture (No. XTE1854). We also would like to express our sincere gratitude to the anonymous reviewers for their careful work and constructive comments that have helped improve the manuscript substantially.
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