470198 Engineering Production of Monomers from the Lysine Pathway

Tuesday, November 15, 2016: 2:00 PM
Continental 6 (Hilton San Francisco Union Square)
Maria McClintock and Kechun Zhang, Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN

There is great interest in developing bio-based aromatic chemicals to replace phthalates, one of the highest volume monomers used in industrial and consumer products. We envision dipicolinic acid (DPA) could be a useful phthalate analog. In addition, the nitrogen atom in the aromatic ring confers new possibilities, as a metal chelator leading to its recent use in specialty polymers, with applications in heavy metal detection1, catalysis2, and even as the primary functional component of a self-repairing polymers3. DPA is naturally produced in Bacillus subtilis cells during sporulation. The biosynthetic pathway, however, has not yet been well-characterized. We have successfully identified the relevant genes for production of DPA in E. coli, including two uncharacterized genes spoVFA and spoVFB which are instrumental for production4. Initial fermentations yielded up to 1.4 g/L production of DPA in 48 hours. The productivity was also examined for E. coli where threonine and lysine production was reduced. Finally, engineered wild-type E. coli was able to produce DPA at maximum of 4.3 g/L with minimal media optimization. DPA production for all fermentations was monitored by HPLC and verified using LC-MS fragmentation. LC-MS of unpurified fermentation media also show production of several by-products, including homoserine and HTPA.
  1. Darbha, G. K., Ray, A. & Ray, P. C. ACS Nano 1, 208–214 (2007).
  2. Li, C.H. et al. Nat. Chem. 1–7 (2016).
  3. Kirillova, M. V., Kirillov, A. M., Guedes Da Silva, M. F. C. & Pombeiro, A. J. L. Eur. J. Inorg. Chem. 3423–3427 (2008).
  4. Daniel, R. A. & Errington, J. J. Mol. Biol. 232, 468–483 (1993).

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