Other

Adelbert Bacher

Petra Adam

Wolfgang Eisenreich

Stefan Hecht

Felix Rohdich

Meinhart H Zenk

Christoph A Schuhr

Monika Fellermeier

Ferdinand Zepeck

Sabine Amslinger

Juraithip Wungsintaweekul

Tobias Gräwert

Assay for testing a sample for the presence or absence of inhibition of the enzymatic conversion of 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphate into isopentenyl diphosphate and/or dimethylallyl diphosphate by the following steps: (a) reacting an aqueous mixture containing 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphate, a 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphnte reductase, and a reducing agent under predetermined reaction conditions for a predetermined period of time; (b) analyzing the reaction mixture obtained in step (a) for the consumed amount of 1-hydroxy-2-methyl-(E)-butenyl 4-diphosphate and/or NAD(P)H and/or for the produced amount of isopentenyl diphosphate, and/or dimethylallyl diphosphate and/or NAD(P); (c) repeating step (a) in the presence of the sample to be tested; (d) repeating step (b) with the reaction mixture defined in step (c); (e) comparing the results of steps (b) and (d).

Assay for inhibitors of ispH

The invention provides a protein in a form that is functional for the enzymatic conversion of 2C-methyl-D-erythritol 2,4-cyclodiphosphate to 1-hydroxy-2-methyl-2-butenyl 4-diphosphate notably in its (E)-form of the non-mevalonate biosynthetic pathway to isoprenoids. The invention also provides a protein in a form that is functional for the enzymatic conversion of 1-hydroxy-2-methyl-2-butenyl 4-diphosphate, notably in its (E)-form, to isopentenyl diphosphate and/or dimethylallyl diphosphate. Further, screening methods for inhibitors of these proteins are provided. Further, 1-hydroxy-2-methyl-2-butenyl 4-diphosphate is provided and chemical and enzymatic methods of its preparation.

Intermediates and enzymes of the non-mevalonate isoprenoid pathway

The present invention is directed to enzymes and intermediates of the non-mevalonate isoprenoid pathway downstream of 2C-methyl-D-erythritol 2,4-cyclopyrophosphate and upstream of isopentenyl pyrophosphate or dimethylallyl pyrophosphate. These are used as a basis for a screening method for inhibitors of these enzymes, and a method for identifying inhibitor-resistant variants thereof. Further disclosures refer to DNA coding for said enzymes and for inhibitor-resistant variants thereof, vectors containing said DNA, cells containing said vector, and plant seeds comprising cells containing said vector. This invention is useful for the inhibition of the biosynthesis of isoprenoids in plants, bacteria and protozoa, for confering herbicide-resistance to plants, as well as for weed control in agriculture using a crop containing a herbicide-resistant gene and an effective amount of a suitable herbicide.

Non-mevalonate isoprenoid pathway

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Kramsach, Austria

Petra Adam