Trehalose 6-phosphate production with energy coupling fermentation by yeast cells

We tried a method for the production of trehalose 6-phosphate (T6P) with energy-coupling fermentation by baker's yeast. T6P was produced in a reaction mixture containing glucose, 5'-UMP, MgSO4, inorganic phosphate, and dried cells of baker's yeast as the enzyme preparation, T6P was isolated from the reaction mixture and identified by TLC, HPLC, GC-MS, and enzymatic methods. The reaction conditions suitable for T6P production were investigated. The formation of T6P and its precursors, glucose 6-phosphate and UDPglucose, at various pHs and concentrations of substrates was examined. Accumulation of T6P was maximum with a reaction mixture containing 1 M glucose, 20 mM 5'-UMP, 20 mM MgSO4, 400 mM sodium phosphate buffer (pH 6.2), and 100 mg/ml dried cells of baker's yeast shaken at 37 degrees C for 6 h. The yield of T6P as a percentage of glucose was 11% (mol/mol) under these reaction conditions.