A mixture of tetracycline
(600 lL, prepared as 10 mg/mL in 70.0% ethanol) and
cycloheximide (450 lL, prepared as 10 mg/mL in distilled
water) was used to prevent microbial contamination during
hydrolysis. Enzyme was added once the mixture reached
50 C and the slurry was stirred at 150 rpm for 48 h