Biosynthesis of allene oxides in Physcomitrella patens

BMC Plant Biology

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.

ISSN: 1471-2229