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WO 2010/128115 Al (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date 11 November 2010 (11.11.2010) WO 2010/128115 Al (51) International Patent Classification: (81) Designated States (unless otherwise indicated, for every C12N 9/16 (2006.01) kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, (21) International Application Number: CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, PCT/EP20 10/056204 DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, (22) International Filing Date: HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, 6 May 2010 (06.05.2010) KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, (25) Filing Language: English NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, (26) Publication Language: English SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (30) Priority Data: 09159520.7 6 May 2009 (06.05.2009) EP (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (71) Applicant (for all designated States except US): GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, NOVOZYMES A/S [DK/DK]; Krogshoejvej 36, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, DK-2880 Bagsvaerd (DK). TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, (72) Inventors; and LV, MC, MK, MT, NL, NO, PL, PT, RO, SE, SI, SK, (75) Inventors/Applicants (for US only): DANIELSEN, Stef- SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, fen [DK/DK]; Dyssegaardsvej 80 A, DK-2870 GW, ML, MR, NE, SN, TD, TG). Dyssegaard (DK). SKOV, Lars Kobberoee [DK/DK]; Aage Berntsens AUe 63, DK-2750 Ballerup (DK). Published: LEITE, Ricardo [PT/PT]; Urbanizacao Monte da Ria, — with international search report (Art. 21(3)) Lt. 14C 3°Dto, P-8005-303 Faro (PT). LAIZE, Vincent [PT/PT]; Rua Sporting Clube Farense 14A - 9°Esq, — with (an) indication(s) in relation to deposited biological P-8000-434 Faro (PT). DA FONSECA, M Leonor Can- material furnished under Rule 13bis separately from the cela [PT/PT]; Rua Joao de Deus, 45 - 3 dto, P-8000-368 description (Rules 13bis.4(d)(i) and 48.2(a)(viii)) Faro (PT). — with sequence listing part of description (Rule 5.2(a)) (74) Common Representative: NOVOZYMES A/S; Krogshoejvej 36, DK-2880 Bagsvaerd (DK). (54) Title: DFPASE ENZYMES FROM OCTOPUS VULGARIS (57) Abstract: The present invention relates to isolated polypeptides having organophosphorous hydrolase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells compris- ing the polynucleotides as well as methods of producing and using the polypeptides. DFPase Enzymes from Octopus Vulgaris Reference to a Sequence Listing This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference. Reference to a Deposit of Biological Material This application contains a reference to a deposit of biological material, which deposit is incorporated herein by reference. For complete information see last paragraph of the description. BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to isolated polypeptides having organophosphorous hydrolase activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides. Description of the Related Art Organophosphorous compounds are known in the art. In particular some warfare agents are known to be organophosphorous compounds such as Sarin, Cyclosarin, and Soman. Other organophosphorous compounds are known as pesticides. It is desirable to be able to decontaminate areas contaminated with such organophosphorous compounds. A polypeptide having organophosphorous hydrolase activity, such as diisopropylfluorophosphatase activity has been suggested for this purpose since such polypeptides are capable of hydrolyzing harmful organophosphorous compounds and thereby converting them to less harmful products. In WO99/43791 a diisopropylfluorophosphatase from Loligo vulgaris is disclosed and its potential use for decontamination among other applications is also described. It is an object of the present invention to provide polypeptides having organophosphorous hydrolase e.g. diisopropylfluorophosphatase activity and polynucleotides encoding the polypeptides, in particular having high stability and /or high specific activity. SUMMARY OF THE INVENTION The present invention relates to isolated polypeptides having organophosphorous hydrolase activity, selected from the group consisting of: (a) a polypeptide comprising an amino acid sequence having at least 75%, more preferably at least 80%, even more preferably at least 85%, most preferably at least 90%, and even most preferably at least 95% identity to the mature polypeptide of SEQ ID NO: 2 ; (b) a polypeptide encoded by a polynucleotide that hybridizes under at least medium-high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) a DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) a full-length complementary strand of (i) or (ii); (c) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 65%, more preferably at least 70%, more preferably at least 75%, more preferably at least 80%, more preferably at least 85%, more preferably at least 90%, more preferably at least 95%, more preferably at least 96%, even more preferably at least 97%, most preferably at least 98% and even most preferably 99% identity to the mature polypeptide coding sequence of SEQ ID NO: 1; (d) a variant comprising a substitution, deletion, and/or insertion of one or more (several) amino acids of the mature polypeptide of SEQ ID NO: 2 . The present invention also relates to isolated polynucleotides encoding polypeptides having organophosphorous hydrolase activity, selected from the group consisting of: (a) a polynucleotide encoding a polypeptide comprising an amino acid sequence having at least 75 % identity to the mature polypeptide of SEQ ID NO: 2 ; (b) a polynucleotide that hybridizes under at least medium stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) a DNA sequence comprising the mature polypeptide coding sequence of SEQ ID NO: 1, or (iii) a full-length complementary strand of (i) or (ii); (c) a polynucleotide comprising a nucleotide sequence having at least 65% identity to the mature polypeptide coding sequence of SEQ ID NO: 1; and (d) a polynucleotide encoding a variant comprising a substitution, deletion, and/or insertion of one or more (several) amino acids of the mature polypeptide of SEQ ID NO: 2 . The present invention also relates to nucleic acid constructs, recombinant expression vectors, recombinant host cells comprising the polynucleotides, and methods of producing a polypeptide having organophosphorous hydrolase activity. The present invention also relates to methods of decontamination, e.g. by degrading organophosphorous compounds. In particular the present invention relates to methods of decontaminating an area or a device contaminated with one or more hazardous or undesired organophosphorous compounds by applying the organophosphorous hydrolase of the invention to said area or device. The present invention also relates to plants comprising an isolated polynucleotide encoding such a polypeptide having organophosphorous hydrolase activity. The present invention also relates to methods of producing such a polypeptide having organophosphorous hydrolase activity, comprising: (a) cultivating a transgenic plant or a plant cell comprising a polynucleotide encoding such a polypeptide having organophosphorous hydrolase activity under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide. Brief Description of the Figures Figure 1 shows the restruction map of NN059107. DEFINITIONS The term "Organophosphorous hydrolase activity" is defined herein as hydrolytic activity to organophosphorous compounds, in particular phosphorous anhydride bonds in organophosphorous compounds including nerve gases. Thus the term includes an enzyme with hydrolase activity and/or esterase activity, e.g. organophosphorous hydrolase activity (EC 3.1 .8.1 ) (such as an organophosphoesterase activity) or organophosphoric acid anhydrolase (OPAA) activity, or carboxylesterase activity, diisopropylfluorophosphatase (DFPase) activity (EC 3.1 .8.2), dehalogenase activity, prolidase activity and/or imidodipeptidase activity. The term "DFPase (EC3. 1.8.2)" is defined herein as diisopropylfluorophosphatase, dialkylfluorophosphatase, diisopropylphosphorofluoridate hydrolase, diisopropylfluorophosphonate dehalogenase, diisopropylphosphofluoridase, isopropylphosphorofluoridase, organophosphate acid anhydrase, organophosphorous acid anhydrolase, somanase, tabunase. DFPases acts on phosphorus anhydride bonds (such as phosphorus-halide and phosphorus-cyanide) in organophosphorous compounds (including nerve gases). The activity of the polypeptides according to the invention is measured as described in Example 4 "Measurement of enzyme activity". The
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