Table 3

Product specificities of different PpAOS1 and PpHPL variants
Substrate Enzyme-variant ω-oxo fatty-acids (= HPL activity) Ketols (= AOS activity) Cyclopentenone (= AO cyclization)
9-HPOD HPL Wt ++++ + n.d.
AOS1 Wt + ++++ n.d.
AOS1 F93L ++++ + n.d.
9-HPOT(n-3) HPL Wt ++++ + n.d.
HPL F151L ++++ + n.d.
HPL A169S ++++ + n.d.
HPL F151L, A169S ++++ + n.d.
AOS1 Wt + ++++ n.d.
AOS1 F93L ++++ - n.d.
13-HPOD HPL Wt ++++ + n.d.
HPL F151L ++++ + n.d.
HPL A169S ++++ + n.d.
HPL F151L, A169S ++++ + n.d.
AOS1 Wt + ++++ n.d.
AOS1 F93L ++++ + n.d.
13-HPOT(n-3) HPL Wt ++++ + n.d.
HPL F151L ++++ + n.d.
HPL A169S ++++ + n.d.
HPL F151L, A169S ++++ + n.d.
AOS1 Wt + +++ +
AOS1 F93L ++ ++ +

Affinity purified enzymes were incubated with [1-14C]-labeled hydroperoxy fatty acids for approx. 30 min. After extraction products were analyzed by RP-HPLC that was coupled to a radio-detector and quantified by integration of the respective peak area. For simplicity the relative amounts of each product is indicated by the number of “+”. AOS, allene oxide synthase; HPL, hydroperoxide lyase; Wt, wild type; n.d., not determined. The data represent between 2 and 5 independent experiments.

Scholz et al.

Scholz et al. BMC Plant Biology 2012 12:228   doi:10.1186/1471-2229-12-228

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