Abstract
The aim of this study was to increase pyruvate production efficiency in a multi-vitamin auxotrophic yeast, Torulopsis glabrata. This was achieved by decreasing intracellular NADH content through the introduction of two different NADH reoxidation pathways: one in the cytoplasm and the other in mitochondria. A nox(encoding a cytoplasmic H2O-forming NADH oxidase) and an AOX1 (encoding a mitochondrial alternative oxidase) were successfully expressed heterologously in T. glabrata, resulting in a decrease in the NADH and ATP contents of 55% and 26%, (in T. glabrata NOX) and 45% and 47% (in T. glabrata AOX), respectively. The decreases in nucleotide concentrations led to increases in the glucose consumption rate, the pyruvate yield and pyruvate productivity of 27%, 15% and 22% (in T. glabrata NOX) and 38%, 21%, and 29% (in T. glabrata AOX), respectively, compared with the corresponding values of the control. We conclude that this method provides an alternative way to enhance the production efficiency of NADH-related metabolites.
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Y. H. Zhu, M. A. Eiteman, R. Altman and E. Altman, Appl. Environ. Microbiol., 74, 6649 (2008).
Y. Li, J. Chen and S.Y. Lun, Appl. Biochem. Biotechnol., 57, 451 (2001).
C. Larsson, A. Nilsson, A. Blomberg and L. Gustafsson, J. Bacteriol., 179, 7243 (1997).
T. B. Causey, K. T. Shanmugam, L. P. Yomano and L. O. Ingram, Proc. Natl. Acad. Sci. USA, 101, 2235 (2004).
L. Pritchard and D. B. Kell, Eur. J. Biochem., 269, 3894 (2002).
G. S. Chuang and M. S. Chiou, Korean J. Chem. Eng., 23, 419 (2006).
G. N. Vemuri, M. A. Eiteman, J. E. McEwen, L. Olsson and J. Nielsen, Proc. Natl. Acad. Sci. USA, 104, 2402 (2007).
K.G. Alberti, J. Clin. Pathol. Suppl. (R Coll Pathol), 11, 14 (1977).
M. Rigoulet, H. Aguilaniu, N. Averet, O. Bunoust, N. Camougrand, X. Grandier-Vazeille, C. Larsson, I. L. Pahlman, S. Manon and L. Gustafsson, Mol. Cell. Biochem., 256–257, 73 (2004).
L. M. Liu, Y. Li, H. Li and J. Chen, FEMS Yeast Res., 6, 1117 (2006).
J.W. Zhou, L. X. Huang, L. M. Liu and J. Chen, J. Biotechnol., 144, 120 (2009).
M. Kanehisa, S. Goto, M. Hattori, K. F. Aoki-Kinoshita, M. Itoh, S. Kawashima, T. Katayama, M. Araki and M. Hirakawa, Nucleic. Acid. Res., 34, D354 (2006).
J. Hou, N. F. Lages, M. Oldiges and G.N. Vemuri, Metab. Eng., 11, 253 (2009).
C. H. Johnson, J. T. Prigge, A. D. Warren and J. E. McEwen, Yeast, 20, 381 (2003).
S. Guerrero-Castillo, M. Vazquez-Acevedo, D. Gonzalez-Halphen and S. Uribe-Carvajal, Biochim. Biophys. Acta, 1787, 75 (2009).
S. Heux, R. Cachon and S. Dequin, Metab. Eng., 8, 303 (2006).
J.W. Zhou, Z. Dong, L. Liu, G. Du and J. Chen, J. Microbiol. Methods, 76, 70 (2009).
L. M. Liu, Y. Li, H. Z. Li and J. Chen, Lett. Appl. Microbiol., 39, 199 (2004).
Y. Qin, Z.Y. Dong, J.W. Zhou, L. M. Liu and J. Chen, Wei Sheng Wu Xue Bao, 49, 1483 (2009).
M. Moreira dos Santos, V. Raghevendran, P. Kotter, L. Olsson and J. Nielsen, Metab. Eng., 6, 352 (2004).
C. Bernofsky and M. Swan, Anal. Biochem., 53, 452 (1973).
M. R. Leonardo, P. R. Cunningham and D. P. Clark, J. Bacteriol., 175, 870 (1993).
L.M. Liu, Y. Li, Z. Shi, G. Du and J. Chen, J. Biotechnol., 126, 173 (2006).
A. Valadi, K. Granath, L. Gustafsson and L. Adler, J. Biol. Chem., 279, 39677 (2004).
E. Postma, C. Verduyn, W. A. Scheffers and J. P. Van Dijken, Appl. Environ. Microbiol., 55, 468 (1989).
A. P. Lin and L. McAlister-Henn, J. Biol. Chem., 277, 22475 (2002).
R. Klassen, J. Fricke, A. Pfeiffer and F. Meinhardt, Biotechnol. Lett., 30, 1041 (2008).
H. Muller, C. Hennequin, J. Gallaud, B. Dujon and C. Fairhead, Eukaryot. Cell., 7, 848 (2008).
K. J. Livak and T. D. Schmittgen, Methods, 25, 402 (2001).
A. L. Moore and J. N. Siedow, Biochim. Biophys. Acta, 1059, 121 (1991).
D. A. Berthold, N. Voevodskaya, P. Stenmark, A. Graslund and P. Nordlund, J. Biol. Chem., 277, 43608 (2002).
D. D. Zhang, N. Liang, Z. P. Shi, L. M. Liu, J. Chen and G. C. Du, Biotechnol. Bioprocess. Eng., 14, 134 (2009).
C. Larsson, I. L. Pahlman and L. Gustafsson, Yeast, 16, 797 (2000).
G. von Jagow and M. Klingenberg, Eur. J. Biochem., 12, 583 (1970).
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Qin, Y., Johnson, C.H., Liu, L. et al. Introduction of heterogeneous NADH reoxidation pathways into Torulopsis glabrata significantly increases pyruvate production efficiency. Korean J. Chem. Eng. 28, 1078–1084 (2011). https://doi.org/10.1007/s11814-010-0483-1
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DOI: https://doi.org/10.1007/s11814-010-0483-1