Metabolomics profiling reveals the mechanism of increased pneumocandin B0 production by comparing mutant and parent strains
- 187 Downloads
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.
- 7.Chen L, Li Y, Yue Q, Loksztejn A, Yokoyama K, Felix EA, Liu XZ, Zhang NY, An ZG, Bills GF (2016) Engineering of new pneumocandin side-chain analogues from Glarea lozoyensis by mutasynthesis and evaluation of their antifungal activity. ACS Chem Biol 11:2724–2733CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Jordan P, Choe JY, Boles E, Oreb M (2016) Hxt13, Hxt15, Hxt16 and Hxt17 from Saccharomyces cerevisiae represent a novel type of polyol transporters. Sci Rep 6:2503Google Scholar
- 20.Kuratsu Y, Arai Y, Inuzuka K, Suzuki T (1983) Stimulatory effect of aspartic-acid on colistin production by Bacillus Polymyxa. Agric Biol Chem Tokyo 47:2607–2612Google Scholar
- 30.Nemeria N, Yan Y, Zhang Z, Brown AM, Arjunan P, Furey W, Guest JR, Jordan F (2001) Inhibition of the Escherichia coli pyruvate dehydrogenase complex E1 subunit and its tyrosine 177 variants by thiamin 2-thiazolone and thiamin 2-thiothiazolone diphosphates. Evidence for reversible tight-binding inhibition. J Biol Chem 276:45969–45978CrossRefPubMedGoogle Scholar
- 33.Qu L, Ren L-J, Li J, Sun G-N, Sun L-N, Ji X-J, Nie Z-K, Huang H (2013) Biomass composition, lipid characterization, and metabolic profile analysis of the fed-batch fermentation process of two different docosahexaenoic acid producing Schizochytrium sp. strains. Appl Biochem Biotechnol 171:1865–1876CrossRefPubMedGoogle Scholar
- 35.Shibukawa M, Ohsawa T (1966) l-Glutamic acid fermentation with molasses: part VI. Effect of the saturated-unsaturated fatty acid ratio in the cell membrane fraction on the extracellular accumulation of l-glutamate. Biosci Biotechnol Biochem 30:750–758Google Scholar