title: Inactivation of pyruvate kinase or the phosphoenolpyruvate: sugar phosphotransferase system increases shikimic and dehydroshikimic acid yields from glucose in Bacillus subtilis.
abstract: The glycolytic intermediate phosphoenolpyruvate (PEP) is a precursor of several cellular components, including various aromatic compounds. Modifications to the PEP node such as PEP:sugar phosphotransferase system (PTS) or pyruvate kinase inactivation have been shown to have a positive effect on aromatics production capacity in Escherichia coli and Bacillus subtilis. In this study, pyruvate kinase and PTS-deficient B. subtilis strains were employed for the construction of derivatives lacking shikimate kinase activity that accumulate two industrially valuable chemicals, the intermediates of the common aromatic pathway, shikimic and dehydroshikimic acids. The pyruvate kinase-deficient strain (CLC6-PYKA) showed the best production parameters under resting-cell conditions. Compared to the PTS-deficient strain, the shikimic and dehydroshikimic acids specific production rates for CLC6-PYKA were 1.8- and 1.7-fold higher, respectively. A batch fermentor culture using complex media supplemented with 83 g/l of glucose was developed with strain CLC6-PYKA, where final titers of 4.67 g/l (shikimic acid) and 6.2 g/l (dehydroshikimic acid) were produced after 42 h.
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Qld., Australia.
authors.forename: Cuauhtemoc
authors.initials: C
authors.lastname: Licona-Cassani
authors.forename: Alvaro R
authors.initials: AR
authors.lastname: Lara
authors.forename: Natividad
authors.initials: N
authors.lastname: Cabrera-Valladares
authors.forename: Adelfo
authors.initials: A
authors.lastname: Escalante
authors.forename: Georgina
authors.initials: G
authors.lastname: Hernández-Chávez
authors.forename: Alfredo
authors.initials: A
authors.lastname: Martinez
authors.forename: Francisco
authors.initials: F
authors.lastname: Bolívar
authors.forename: Guillermo
authors.initials: G
authors.lastname: Gosset
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways / Q000235:genetics; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000037:antagonists & inhibitors* / Q000235:genetics; D011770:Pyruvate Kinase / Q000037:antagonists & inhibitors* / Q000235:genetics; D012765:Shikimic Acid / Q000031:analogs & derivatives* / Q000378:metabolism*
chemical_list: C096387:3-dehydroshikimate; D012765:Shikimic Acid; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; D011770:Pyruvate Kinase