Linoleate and linolenate-lipoxygenase mutants

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Plant proteins – e.g. – derived from legumes – algae or...

Reexamination Certificate

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Reexamination Certificate

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06875847

ABSTRACT:
The present invention concerns a procedure for the manufacture of plant lipoxygenases with altered positional specificity as well as the lipoxygenase obtained through the procedure and its use for the hydroperoxylation of substrates. In particular the LOXs according to the invention permit for the first time the manufacture of new γ-linolenic acid derivatives in large quantity. For this γ-linolenic acid as substrate is incubated under suitable conditions with the LOXs according to the invention. Depending on the mutant LOXs put into use a hydroperoxylation of the γlinolenic acid then ensues, preferably at position 6 or position 9 or positions 6 and 9.

REFERENCES:
Feussner et al., “All three acyl moieties of trilinolein are efficiently oxygenated by recombinant His-tagged lipid body lipoxygenase in vitro,”FEBS Letters, 431(3), 433-436 (Jul. 24, 1998).
Hornung et al., “Conversion of cucumber linoleate 13-lipoxygenase to a 9-lipoxygenating species by site-directed mutagenesis,”PNAS USA, 96(7), 4192-4197 (Mar. 30, 1999).
Patent Abstracts of Japan, 012 (338) (C-527) (Sep. 12, 1988) & JP 63-098392 A (Agency of Ind. Science & Technology) (Apr. 28, 1988).
Patent Abstracts of Japan, 014 (497) (C-0774) (Oct. 30, 1990) & JP 02-207792 (Agency of Ind. Science & Technology) (Aug. 17, 1990).
Prigge et al., “Structure conservation in lipoxygenases: structural analysis of soybean lipoxygenase-1 and modeling of human lipoxygenases,”Proteins, 24(3), 275-291, (Mar., 1996).
Sloane et al., “Conversion of human 15-lipoxygenase to an efficient 12-lipoxygenase: the side-chain geometry of amino acids 417 and 418 determine positional specificity,”Protein Engineering, 8(3), 275-282 (Mar., 1995).
Steczko et al., “Conserved histidine residues in soybean lipoxygenase: functional consequences of their replacement,”Biochemistry, 31(16), 4053-4057 (Apr. 28, 1992).

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