Studies on biosynthesis of waxes by developing jojoba seed. II. the demonstration of wax biosynthesis by cell-free homogenates

Abstract

The following enzyme activities were demonstrated in cell-free homogenates from developing jojoba cotyledons: 1) elongation of long chain acyl-CoAs in the presence of malonyl-CoA and NADPH (or NADH), 2) NADPH-dependent reduction of long chain acyl-CoAs to the corresponding alcohols, 3) esterification of long chain acyl-CoAs and the alcohols produced from them into wax, 4) elongation of stearoyl-ACP to eicosanoate and docosanoate as well as reduction to stearyl alcohol, 5) desaturation of stearoyl-ACP to oleate in the presence of reduced ferredoxin, and 6) incorporation of malonyl-CoA into long chain fatty acids and alcohols in the presence of added acyl carrier protein. These activities were associated entirely with the floating wax pad after centrifugation of the cell-free homogenate at 12,000 g for 20 min. The relevance of the above reactions (1–6) to wax biosynthesis in vivo is discussed. Production of oleate from acetate by enzymes utilizing ACP-thioesters as substrates followed by conversion of oleyl-ACP to oleoyl-CoA (via free oleic acid) for subsequent elongation, reduction, and esterification, is presented as the most probable in vivo pathway, for wax biosynthesis. The substrate specificities of the elongation and reduction reactions utilizing acyl-CoAs as substrates are examined in terms of wax composition.