Comparison between typical and screened reaction conditions to optimize cost, efficiency, and yield of milk thistle glucuronides

ReagentTypical ConditionaScreened Condition(s)
Tris-HCl (pH 7.4) (mM)b100100
MgCl2 (mM)b55
BSA (%, w/v)b0.50.5
Alamethicin (μM)b2525
Silybin A (mM)b0.40.4
UDPGA (mM)4.04.0, 0.8
Enzyme sourceHLMsBLMs, bovine S9 fraction
Enzyme source concentration (mg/ml protein)1.001.00, 0.50, 0.25 (BLMs); 2.00, 1.00, 0.50 (S9)
  • a Walker et al. (2014).

  • b Concentration was not varied to limit experimental complexity.