The progress of the NMPS×s and NMP×

The progress of the NMPS×s and NMP×s synthesis with the use of the whole E.
herbicola cells was monitored by HPLC and TLC [9]. The reaction mixture (1 ml) containing donor (10 m
) of
the thiophosphate group, AMPS, or phosphate group, AMP, nucleoside acceptor (50 m
), sodium acetate
buffer (0.2
, pH 4.5), and wet paste of intact E. herbicola cells (6.0 mg, calculated as abs. dry weight) was
incubated at 50 with gentle stirring. Aliquots of the reaction medium were analyzed by the TLC on Silufol UV-
254 plates (Serva, Germany; i-PrOH/25% aq. NH3/H2O 7 :2 :1 (v/v)) and/or by HPLC with a molar extinction
coefficients for long-wave maximum for nucleoside under investigation. The amount of the enzyme producing
1 nmol of NMPS per min was taken to be one unit.