not specified

The culture broth with a volume corresponding to about 100 mg (dry weight) of cells was centrifuged (4 °C, 10 min at 10,000×g) and washed twice with 20 mM Tris-HCl buffer (pH at 7.0). Cells were resuspended in Tris (100 mM) -HCl buffer (pH at 7.0) containing KCl (20 mM), MnS ... O4 (5 mM), DTT (2 mM) and EDTA (0.1 mM). Cell disruption by sonication (five cycles of 30 s with 1 min cooling periods) was followed by the removal of cell debris by centrifugation for 10 min at 10,000×g at 4 °C. The supernatant was used for all enzyme assays. The protein concentration of extracts was determined by the Lowry's method [1] with bovine serum albumin used as the standard.
Enzyme assays were made based on the relationships between the enzyme activity and the consumption or production of NADH or NADPH monitored at 340 nm in a spectrophotometer. The enzyme activities of glucose 6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) were measured based on the methods of Moritz et al. [2]. The enzyme activities of Pyk, lactate dehydrogenase (LDH), Ack, Ppc and alcohol dehydrogenase (ADH) were measured by Riondet et al.'s method [3]. The enzyme activity of GAPDH was measured based on the method of [4]. The enzyme activity of pyruvate-formate-lyase (Pfl) was measured based on Garrigues et al. [5].
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[1] O.H. Lowry, N.J. Rosebrough, A.L. Farr, R.J. J. Biol. Chem., 193 (1951), pp. 265-275.
[2] B. Moritz, K. Striegel, A.A. de Graaf, H. Sahm. Eur. J. Biochem., 267 (2000), pp. 3442-3452. http://doi.org/fwnmpm
[3] C. Riondet, R. Cachon, Y. Waché, G. Alcaraz, C. Diviès. J. Bacteriol., 182 (2000), pp. 620-626. http://doi.org/cf5kb7
[4] S. Even, C. Garrigues, P. Loubiere, N.D. Lindley, M. Cocaign-Bousquet. Metab. Eng., 1 (1999), pp. 198-205. http://doi.org/b23hk8
[5] C. Garrigues, P. Loubiere, N.D. Lindley, M. Cocaign-bousquet. J. Bacteriol., 179 (1997), pp. 5282-5287. http://doi.org/cscc