Sucrose → L-Ribose

Participants

Reaction Steps

Cellulomonas gerida (JCM1490) strain was inoculated at a concentration of 1% to 18 L of a medium containing 2.0 g/L of sucrose, 5.0 g/L of yeast extract, 5.0 g/L of soybean peptides, 5 g/L of NaCl, 3 g/L of K2HPO4, 1 g/L of KH2PO4 and 15 g/L of L-ribose (pH 7.0) contained in a 30-L jar. The seed culture was obtained beforehand by culturing the bacterium in the same medium contained in a flask with a baffle at 30° C. for 18 hours with shaking at 160 rpm. The conditions in the jar were controlled to be 300 rpm, 1 vvm and 30° C., and the culture was performed for 18 hours. The obtained culture broth in which the culture was completed was centrifuged to collect the cells. The cell pellet was added with 1.8 L of glycine-HCl buffer (50 mM, pH 8.5) and homogenously suspended in the buffer. Then, the suspension was added with 60 mL of toluene and stirred vigorously for 15 minutes. 1.2 L of the toluene-treated cells prepared as described above was added to 12.5 L of the crude L-ribulose solution obtained in 2), and the reaction was performed at 30° C. for 24 hours while gently stirring the mixture so that the mixture should become uniform. During the reaction, pH was controlled to be 8.5 with 1 N NaOH. The obtained mixture after the reaction was centrifuged to remove the cells, and passed through a precision membrane filter with a pore size of 0.22 μm to obtain 12 L of transparent crude L-ribose solution. The concentration of L-ribose was 30 g/L, and the concentration of L-ribulose was 13 g/L.

Provenance & Evidence