Fig. 5
Biochemical characterization of the jojoba seed lipase activity
. a. The impact of temperature on the jojoba seed lipase activity towards 18:1-TAG (left panel) and 18:1–18:1 WE (right panel). Data represent mean values and error bars show the range of duplicates. Assay conditions: aliquots (2.5 nmol of endogenous PC) of microsomal fraction mixture isolated from two individual jojoba seeds from each accession (35 dpg); 20 nmol of [14C]18:1-TAG or [14C]18:1–18:1 WE added to freeze-dried microsomes in 19 μl benzene; benzene evaporation, and addition of 100 μl 0.1 HEPES buffer (pH 7.0); incubation for 2.5 min (for TAG) or 1.5 min (for WE). b. The impact of the pH on the jojoba seed lipase activity towards 18:1-TAG (left panel) and 18:1–18:1 WE (right panel). Data represent mean values and error bars show the range of duplicates. Assay condition: as above with 0.1 M citrate buffer (pH 4.0–5.0), phosphate buffer (pH 6.0–8.0), or Tris-HCl buffer (9.0), 5 min incubation at 35 °C. c. The impact of pre-incubation conditions on jojoba seed lipase activity towards 18:1-TAG. Data represent mean values and error bars show the range of duplicates; nd – not detected. Assay conditions: as above, 5 min incubation at 60 °C. d. Substrate specificity of jojoba seed lipases towards saturated and unsaturated WEs. Data represent mean values and error bars show the range of duplicates. Assay condition: as above with 2.5 nmol of [14C] WE, 10 min incubation at 60 °C.