Fig. 4.

One-step preparative RP-HPLC purification of TM 1–99. Conditions: linear AB gradient (Eluent A is 0.05% aq. TFA, pH 2.0 and Eluent B is 0.05% TFA in acetonitrile) of 2% B/min up to 24% B, followed by a gradient of 0.1%/min up to 40% B. A gradient of 4% B/min up to 60% B was then followed by an isocratic wash with 60% B; flow-rate, 0.3 ml/min; temperature, 25 °C. 2 min fractions were collected and subjected to the same analytical conditions as Fig. 1 except for a gradient rate of 2%B/min. Sample volume: 4 ml, containing 23 mg of lyophilized crude cell extract. P denotes desired product.