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Manganese orchestrates a metabolic shift leading to the increased bioconversion of glycerol into α-ketoglutarate.

Faculty of Science and Engineering, Laurentian University, Sudbury, ON, P3E 2C6, Canada

Glycerol is a major by-product of the biodiesel industry and its transformation into value-added products is an ongoing technological challenge. Here we report on the ability of the nutritionally-versatile Pseudomonas fluorescens to synthesize copious amount of α-ketoglutarate (KG) in a glycerol medium supplemented with manganese (Mn). The enhanced production of this keto-acid was mediated by the increased activities of isocitrate dehydrogenase (ICDH)-(NAD)P dependent and aminotransaminases. At stationary phase of growth when the optimal quantity of KG was recorded, these enzymes exhibited maximal activities. Two isoforms of pyruvate carboxylase (PC) that were identified in the Mn-treated cells provided an effective route for the synthesis of oxaloacetate, a metabolite critical in the production of KG. Furthermore, the increased activities of phosphoenol pyruvate carboxylase (PEPC) and pyruvate orthophosphate dikinase (PPDK) ensured the efficacy of this KG-generating metabolic system by supplying pyruvate and ATP from the oxaloacetate synthesized by PC. Mn-exposed whole cells converted 90% of industrial glycerol into KG. This Mn-evoked metabolic network can be optimized into the economic transformation of glycerol into KG.
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1. Kumar M, Gayen K (2011) Developments in biobutanol production: new insights. Appl Ener 88: 1999–2012.

2. Hoekman SK (2009) Biofuels in the US-challenges and opportunities. Renew Ener 34: 14–22.

3. Behera S, Singh R, Arora R, et al. (2014) Scope of algae as third generation biofuels. Front Bioeng Biotechnol 2: 90.

4. Saxena RC, Adhikari DK, Goyal HB (2009) Biomass-based energy fuel through biochemical routes: a review. Renew Sust Energ Rev 13: 167–178.    

5. Ullah K, Ahmad M, Sharma VK, et al. (2014) Algal biomass as a global source of transport fuels: overview and development perspectives. Prog Nat Sci Mater Int 24: 329–339.    

6. Elkins JG, Raman B, Keller M (2010) Engineered microbial systems for enhanced conversion of lignocellulosic biomass. Curr Opin Biotechnol 21: 657–662.    

7. Papagianni M (2012) Recent advances in engineering the central carbon metabolism of industrially important bacteria. Microb Cell Fact 11: 1–13.    

8. Liao J, Mi L, Pontrelli S, et al. (2016) Fuelling the future: microbial engineering for the production of sustainable biofuels. Nat Rev Microbiol 14: 288–304.

9. Suero SR, Ledesma B, Álvarez-Murillo A, et al. (2015) Glycerin, a biodiesel by-product with potentiality to produce hydrogen by steam gasification. Energies 8: 12765–12775.    

10. Luna C, Verdugo C, Sancho ED, et al. (2014) Production of a biodiesel-like biofuel without glycerol generation, by using Novozym 435, an immobilized Candida antarctica lipase. BRBP 1: 1–11.

11. Da Silva GP, Mack M, Contiero J (2009) Glycerol: a promising and abundant carbon source for industrial microbiology. Biotechnol Adv 27: 30–39.    

12. Bagheri S, Julkapli NM, Yehye WA (2015) Catalytic conversion of biodiesel derived raw glycerol to value added products. Energy Rev 41: 113–127.

13. Johnson DT, Taconi KA (2007) The glycerin glut: options for the value‐added conversion of crude glycerol resulting from biodiesel production. Environ Prog 26: 338–348.    

14. Fan X, Chen R, Chen L, et al. (2016) Enhancement of alpha-ketoglutaric acid production from L-glutamic acid by high-cell-density cultivation. J Mol Catal B: Enzym 126: 10–17.    

15. Appanna VD (1988) Alteration of exopolysaccharide composition in Rhizobium meliloti JJ-1 exposed to manganese. Fems Microbiol Lett 50: 141–144.    

16. Kehres DG, Maguire ME (2003) Emerging themes in manganese transport, biochemistry and pathogenesis in bacteria. Fems Microbiol Rev 27: 263–290.    

17. Jakubovics NS, Jenkinson HF (2001) Out of the iron age: new insights into the critical role of manganese homeostasis in bacteria. Microbiology 147: 1709–1718.    

18. Culotta VC, Daly MJ (2013) Manganese complexes: diverse metabolic routes to oxidative stress resistance in prokaryotes and yeast. Antioxid Redox Signal 19: 933–944.    

19. Appanna VD (1988) Stimulation of exopolysaccharide production in Rhizobium meliloti JJ-1 by manganese. Biotechnol Lett 10: 205–206.    

20. Appanna VD, Preston M (1987) Manganese elicits the synthesis of a novel exopolysaccharide in an arctic Rhizobium. Febs Let 215: 79–82.    

21. Otto C, Yovkova V, Barth G (2011) Overproduction and secretion of α-ketoglutaric acid by microorganisms. Appl Microbiol Biotechnol 92: 689–695.    

22. Anderson S, Appanna VD, Huang J, et al. (1992) A novel role for calcite in calcium homeostasis. Febs Let 308: 94–96.    

23. Hamel R, Appanna VD (2003) Aluminum detoxification in Pseudomonas fluorescens is mediated by oxalate and phosphatidylethanolamine. BBA 1619: 70–76.

24. Hamel R, Appanna VD, Viswanatha T, et al. (2004) Overexpression of isocitrate lyase is an important strategy in the survival of Pseudomonas fluorescens exposed to aluminum. Biochem Biophys Res Commun 317: 1189–1194.    

25. Appanna VD, Gazs LG, Pierre MS (1996) Multiple-metal tolerance in Pseudomonas fluorescens and its biotechnological significance. J Biotechnol 52: 75–80.    

26. Bradford MM (1976) A Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254.    

27. Alhasawi A, Leblanc M, Appanna ND, et al. (2015) Aspartate metabolism and pyruvate homeostasis triggered by oxidative stress in Pseudomonas fluorescens: a functional metabolomics study. Metabolomics 11: 1792–1801.    

28. Frank J, Pompella A, Biesalski HK (2000) Histochemical visualization of oxidant stress. Free Radical Biol Med 29: 1096–1105.    

29. Kuhn J, Müller H, Salzig D, et al. (2015) A rapid method for an offline glycerol determination during microbial fermentation. Electr J Biotechnol 18: 252–255.    

30. Middaugh J, Hamel R, Jean-Baptiste G, et al. (2005) Aluminum triggers decreased aconitase activity via Fe-S cluster disruption and the overexpression of isocitrate dehydrogenase and isocitrate lyase: a metabolic network mediating cellular survival. J Biol Chem 280: 3159–3165.    

31. Schägger H, von Jagow G (1991) Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal Biochem 199: 223–231.    

32. Auger C, Appanna VD (2015) A novel ATP-generating machinery to counter nitrosative stress is mediated by substrate-level phosphorylation. BBA 1850: 43–50.

33. Auger C, Lemire J, Cecchini D, et al. (2011) The metabolic reprogramming evoked by nitrosative stress triggers the anaerobic utilization of citrate in Pseudomonas fluorescens. Plos One 6: e28469.    

34. Alhasawi A, Castonguay Z, Appanna ND, et al. (2015) Glycine metabolism and anti-oxidative defence mechanisms in Pseudomonas fluorescens.Microbiol Res 171: 26–31.    

35. Lietzan AD, Maurice MS (2013) Insights into the carboxyltransferase reaction of pyruvate carboxylase from the structures of bound product and intermediate analogs. Biochem Biophys Res Commun 441: 377–382.    

36. Bignucolo A, Appanna VP, Thomas SC, et al. (2013) Hydrogen peroxide stress provokes a metabolic reprogramming in Pseudomonas fluorescens: enhanced production of pyruvate. J Biotechnol 167: 309–315.    

37. Auger C, Appanna V, Castonguay Z, et al. (2012) A facile electrophoretic technique to monitor phosphoenolpyruvate‐dependent kinases. Electrophoresis 33: 1095–1101.    

38. Singh R, Lemire J, Mailloux RJ, et al. (2009) An ATP and oxalate generating variant tricarboxylic acid cycle counters aluminum toxicity in Pseudomonas fluorescens. Plos One 4: e7344.    

39. Whittaker JW (2002) Prokaryotic manganese superoxide dismutases. Methods Enzymol 349: 80–90.    

40. Igarashi T, Kono Y, Tanaka K (1996) Molecular cloning of manganese catalase from Lactobacillus plantarum. J Biol Chem 271: 29521–29524.    

41. Tseng HJ, Srikhanta Y, McEwan AG, et al. (2000) Accumulation of manganese in Neisseria gonorrhoeae correlates with resistance to oxidative killing by superoxide anion and is independent of superoxide dismutase activity. Mol Microbiol 40: 1175–118.

42. Zeczycki TN, Menefee AL, Jitrapakdee S, et al. (2011) Activation and inhibition of pyruvate carboxylase from Rhizobium etli. Biochem 50: 9694–9707.    

43. Adina-Zada A, Jitrapakdee S, Wallace JC, et al. (2014) Coordinating role of His216 in MgATP binding and cleavage in pyruvate carboxylase. Biochem 53: 1051–1058.    

44. Yovkova V, Otto C, Aurich A, et al. (2014) Engineering the α-ketoglutarate overproduction from raw glycerol by overexpression of the genes encoding NADP-dependent isocitrate dehydrogenase and pyruvate carboxylase in Yarrowia lipolytica. Appl Microbiol Biotechnol 98: 2003–2013.    

45. Lietzan AD, Maurice MS (2013) A substrate-induced biotin binding pocket in the carboxyltransferase domain of pyruvate carboxylase. J Biolog Chem 288: 19915–19925.    

46. Li Q, Chen LS, Jiang HX, et al. (2010) Effects of manganese-excess on CO2 assimilation, ribulose-1, 5-bisphosphate carboxylase/oxygenase, carbohydrates and photosynthetic electron transport of leaves, and antioxidant systems of leaves and roots in Citrus grandis seedlings. BMC Plant Boil 10: 42–10.    

47. Mailloux RJ, Puiseux-Dao S, Appanna VD (2009) α-ketoglutarate abrogates the nuclear localization of HIF-1α in aluminum-exposed hepatocytes. Biochimie 91: 408–415.    

48. Husain A, Sato D, Jeelani G, et al. (2012) Dramatic increase in glycerol biosynthesis upon oxidative stress in the anaerobic protozoan parasite Entamoeba histolytica. Plos Negl Trop Dis 6: e1831.    

49. Kimura M, Ujihara M, Yokoi K (1996) Tissue manganese levels and liver pyruvate carboxylase activity in magnesium-deficient rats. Biol Trace Elem Res 52: 171–179.    

Copyright Info: © 2017, Vasu D. Appanna, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

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