Pny326

Brevibacillus choshinensis (Takara Bio Inc.) was used to express phospholipase D. Plasmid pNY326 and pNCMO2 (Takara Bio Inc.) were used as the expression vector. The strain E. coli JM109 (Takara Bio Inc.) was used as the cloning host. Other chemicals, unless otherwise noted, were purchased from Sinopharm Chemical Reagent Co. Ltd. (SCRC; Shanghai, China) Cultivation medium and conditions E. coli JM109 was cultured on the Luria broth (LB) medium containing 10.0 g/L tryptone, 5.0 g/L yeast extract, and 10.0 g/L NaCl, with the introduction of 50 mg/L Ampicillin (Amp) to each culture. In the shake-ask culture, the primary inoculum of B. choshinensis was grown for 24 h in the 250-mL ba ed asks containing 50 mL of TM medium at 30 ℃ and 120 rpm in a shaker until OD 600 = 2~3.
Subsequently, the seed culture (0.5 mL) was diluted to 50 mL of medium A in a 250-mL shake ask at 30 ℃ and 120 rpm. The TM medium contained 10.0 g/L glucose, 10.0 g/L tryptone, 5.0 g/L beef extract, 2.0 g/L yeast extract, 10.0 mg/L FeSO