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Metabolomics analysis of the effect of dissolved oxygen on spinosad production by Saccharopolyspora spinosa

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Abstract

Spinosad, a universal bio-pesticide, is obtained from the soil actinomycete Saccharopolyspora spinosa. Dissolved oxygen, an important contributing factor in aerobic microbial fermentation, however, is not always available in sufficient amounts. To alleviate oxygen limitation in spinosad production, three different oxygen vectors, namely oleic acid, toluene, and n-dodecane, were added into early fermentation. Results indicated that n-dodecane was the optimal oxygen vector. Spinosad yield was increased by 44.2% compared to that in the control group in the presence of 0.5% n-dodecane, added after 120 h of incubation. Yields of the test group reached 6.52 mg/g dry cell weight (DCW), while that of the control group was limited to 4.52 mg/g DCW. Metabolomics analysis by gas chromatography coupled to mass spectrometry was performed to demonstrate the metabolism mechanism in the presence and absence of oxygen vector. In total, 78 principal intracellular metabolites in S. spinosa were detected and quantified in the presence and absence of n-dodecane. Levels of some metabolites that were related to the tricarboxylic acid cycle and pentose phosphate pathway varied significantly. Aspartic acid and glucose-1-phosphate levels varied significantly and contributed most in the distinction of the fermentation conditions and phases. The above findings give new insights into the improvement and the metabolomic characteristics of industrial spinosad production.

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References

  • Devantier R, Scheithauer B, Villas-Bôas SG, Pedersen S, Olsson L (2005) Metabolite profiling for analysis of yeast stress response during very high gravity ethanol fermentations. Biotechnol Bioeng 90:703–714

    Article  CAS  PubMed  Google Scholar 

  • Ding MZ, Cheng JS, Xiao WH, Qiao B, Yuan YJ (2009a) Comparative metabolomic analysis on industrial continuous and batch ethanol fermentation processes by GC-TOF-MS. Metabolomics 5:229–238

    Article  CAS  Google Scholar 

  • Ding MZ, Zhou X, Yuan YJ (2009b) Metabolome profiling reveals adaptive evolution of Saccharomyces cerevisiae during repeated vacuum fermentations. Metabolomics 6:42–55

    Article  Google Scholar 

  • Dripps JE, Boucher RE, Chloridis A, Cleveland CB, DeAmicis CV, Gomez LE, Paroonagian DL, Pavan LA, Sparks TC, Watson GB (2011) Green trends in insect control. In: Lopez O (ed) The spinosad, insecticides. Royal Society of Chemistry, Cambridge, pp 163–212

    Google Scholar 

  • Fiehn O, Kopka J, Dormann P, Altmann T, Trethewey RN, Willmitzer L (2000) Metabolite profiling for plant functional genomics. Nat Biotechnol 18:1157–1161

    Article  CAS  PubMed  Google Scholar 

  • Folescu E, Blaga AC (2013) Utilization of olive oil as a potential oxygen-vector in stirred bioreactors. Environ Eng Manag J 12:587–594

    CAS  Google Scholar 

  • Han L, Mouncey N (2014) Enhancing spinosyn production with oxygen binding proteins. US Patent 8741603

  • Hertlein MB, Mavrotas C, Jousseaume C, Lysandrou M, Thompson GD, Jany W, Ritchie SA (2010) A review of spinosad as a natural product for larval mosquito control. J Am Mosq Control Assoc 26:67–87

    Article  CAS  PubMed  Google Scholar 

  • Hodgson DA (2000) Primary metabolism and its control in streptomycetes: a most unusual group of bacteria. Adv Microb Physiol 42:47–238

    Article  CAS  PubMed  Google Scholar 

  • Huang K, Xia L, Zhang Y, Ding X, Zahn JA (2009) Recent advances in the biochemistry of spinosyns. Appl Microbiol Biot 82:13–23

    Article  CAS  Google Scholar 

  • Krebs HA (1970) Rate control of the tricarboxylic acid cycle. Adv Enzyme Regul 8:335–353

    Article  CAS  PubMed  Google Scholar 

  • Li CX, Florova G, Akopiants K, Reynolds KA (2004) Crotonyl-coenzyme A reductase provides methylmalonyl-CoA precursors for monensin biosynthesis by Streptomyces cinnamonensis in an oil-based extended fermentation. Microbiology 150:3463–3472

    Article  CAS  PubMed  Google Scholar 

  • Li MH, Meng XM, Diao EJ, Du FL, Zhao XY (2012) Productivity enhancement of S-adenosylmethionine in Saccharomyces cerevisiae using n-hexadecane as oxygen vector. J Chem Technol Biot 87:1379–1384

    Article  CAS  Google Scholar 

  • Liang Y, Lu WY, Wen JP (2009) Improvement of Saccharopolyspora spinosa and the kinetic analysis for spinosad production. Appl Biochem Biotech 152:440–448

    Article  CAS  Google Scholar 

  • Luo YS, Kou XX, Ding XZ, Hu SB, Tang Y, Li WP, Huang F, Yang Q, Chen HN, Xia LQ (2012) Promotion of spinosad biosynthesis by chromosomal integration of the Vitreoscilla hemoglobin gene in Saccharopolyspora spinosa. Sci China Life Sci 55:172–180

    Article  CAS  PubMed  Google Scholar 

  • Madduri K, Waldron C, Matsushima P, Broughton MC, Crawford K, Merlo DJ, Baltz RH (2001) Genes for the biosynthesis of spinosyns: applications for yield improvement in Saccharopolyspora spinosa. J Ind Microbiol Biot 27:399–402

    Article  CAS  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Ossipov V, Ossipova S, Bykov V, Oksanen E, Koricheva J, Haukioja E (2008) Application of metabolomics to genotype and phenotype discrimination of birch trees grown in a long-term open-field experiment. Metabolomics 4:39–51

    Article  CAS  Google Scholar 

  • Pan HX, Li JA, He NJ, Chen JY, Zhou YM, Shao L, Dj Chen (2011) Improvement of spinosad production by overexpression of gtt and gdh controlled by promoter PermE* in Saccharopolyspora spinosa SIPI-A2090. Biotechnol Lett 33:733–739

    Article  CAS  PubMed  Google Scholar 

  • Pham TK, Chong PK, Gan CS, Wright PC (2006) Proteomic analysis of Saccharomyces cerevisiae under high gravity fermentation conditions. J Proteome Res 5:3411–3419

    Article  CAS  PubMed  Google Scholar 

  • Reeves AR, Brikun IA, Cernota WH, Leach BI, Gonzalez MC, Weber JM (2007) Engineering of the methylmalonyl-CoA metabolite node of Saccharopolyspora erythraea for increased erythromycin production. Metab Eng 9:293–303

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roessner U, Luedemann A, Brust D, Fiehn O, Linke T, Willmitzer L, Ferniea A (2001) Metabolic profiling allows comprehensive phenotyping of genetically or environmentally modified plant systems. Plant Cell 13:11–29

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sparks TC, Crouse GD, Durst G (2001) Natural products as insecticides: the biology, biochemistry and quantitative structure–activity relationships of spinosyns and spinosoids. Pest Manag Sci 57:896–905

    Article  CAS  PubMed  Google Scholar 

  • Strobel RJ, Nakatsukasa WM (1993) Response surface methods for optimizing Saccharopolyspora spinosa, a novel macrolide producer. J Ind Microbiol 11:121–127

    Article  CAS  Google Scholar 

  • Tang Y, Xia L, Ding X (2011) Molecular cloning and overexpression of partial spinosyn biosynthetic gene cluster in Saccharopolyspora spinosa enhances spinosad production. FEMS Microbiol Lett 325:22–29

    Article  CAS  PubMed  Google Scholar 

  • Villas-Boas SG, Delicado DG, Akesson M, Nielsen J (2003) Simultaneous analysis of amino and nonamino organic acids as methyl chloroformate derivatives using gas chromatography-mass spectrometry. Anal Biochem 322:134–138

    Article  CAS  PubMed  Google Scholar 

  • Wagner C, Sefkow M, Kopka J (2003) Construction and application of a mass spectral and retention time index database generated from GC/EI-TOF-MS metabolite profiles. Phytochemistry 62:887–900

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Xue W, He YM, Peng RH, Yao QH (2014a) Improvement of the ability to produce spinosad in Saccharopolyspora spinosa through the acquisition of drug resistance and genome shuffling. Ann Microbiol 65:771–777

    Article  Google Scholar 

  • Wang XY, Zhang CB, Wang ML, Lu WY (2014b) Genome-scale metabolic network reconstruction of Saccharopolyspora spinosa for Spinosad Production improvement. Microb Cell Fact 13:41

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Williams T, Valle J, Vinuela E (2003) Is the naturally derived insecticide Spinosad® compatible with insect natural enemies. Biocontrol Sci Technol 13:459–475

    Article  Google Scholar 

  • Xue CY, Duan YJ, Zhao FL, Lu WY (2013a) Stepwise increase of spinosad production in Saccharopolyspora spinosa by metabolic engineering. Biochem Eng J 72:90–95

    Article  CAS  Google Scholar 

  • Xue CY, Zhang XM, Yu ZR, Zhao FL, Wang ML, Lu WY (2013b) Up-regulated spinosad pathway coupling with the increased concentration of acetyl-CoA and malonyl-CoA contributed to the increase of spinosad in the presence of exogenous fatty acid. Biochem Eng J 81:47–53

    Article  CAS  Google Scholar 

  • Yang W, Bernards MA (2007) Metabolite profiling of potato (Solanum tuberosum L.) tubers during wound-induced suberization. Metabolomics 3:147–159

    Article  CAS  Google Scholar 

  • York JD (2006) Regulation of nuclear processes by inositol polyphosphates. BBA-Mol Cell Biol L 1761:552–559

    CAS  Google Scholar 

  • Zhang XM, Xue CY, Zhao FL, Li DS, Yin J, Zhang CB, Cai YQ, Lu WY (2014) Suitable extracellular oxidoreduction potential inhibit rex regulation and effect central carbon and energy metabolism in Saccharopolyspora spinosa. Microb Cell Fact 13:98

    PubMed  PubMed Central  Google Scholar 

  • Zhao FL, Xue CY, Wang ML, Wang XY, Lu WY (2013) A comparative metabolomics analysis of Saccharopolyspora spinosa WT, WH124, and LU104 revealed metabolic mechanisms correlated with increases in spinosad yield. Biosci Biotechnol Biochem 77:1661–1668

    Article  CAS  PubMed  Google Scholar 

  • Zhu P, Dong SH, Li S, Xu XY, Xu H (2014) Improvement of welan gum biosynthesis and transcriptional analysis of the genes responding to enhanced oxygen transfer by oxygen vectors in Sphingomonas sp. Biochem Eng J 90:264–271

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by the Project Supported by the Natural Science Foundation of China (No. 21076148 and 31270087), and the Plan for Tianjin Science and Technology Support (No. 11ZCKFSY0100).

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Correspondence to Wenyu Lu.

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The authors declare that they have no conflict of interest.

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Chunzhe Lu and Jing Yin have contributed equally to this work.

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Lu, C., Yin, J., Zhao, F. et al. Metabolomics analysis of the effect of dissolved oxygen on spinosad production by Saccharopolyspora spinosa . Antonie van Leeuwenhoek 110, 677–685 (2017). https://doi.org/10.1007/s10482-017-0835-5

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  • DOI: https://doi.org/10.1007/s10482-017-0835-5

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