MM WIDDO A LITERATURAUS009790528B2 ATARIATO TELUT (12 ) United States Patent ( 10 ) Patent No. : US 9 , 790 ,528 B2 Panaccione et al. (45 ) Date of Patent : Oct . 17 , 2017 (54 ) PRODUCTION OF LYSERGIC ACID BY FOREIGN PATENT DOCUMENTS GENETIC MODIFICATION OF A FUNGUS WO 2014 / 030096 A2 2 /2014 (71 ) Applicant : West Virginia University , Morgantown , WV (US ) OTHER PUBLICATIONS ( 72 ) Inventors: Daniel G . Panaccione , Morgantown , Notification of Transmittal of the International Search Report WV (US ) ; Sarah L . Robinson , ( Forms PCT/ ISA / 220 and PCT/ ISA / 210 ) and the Written Opinion Independence , WV (US ) of the International Searching Authority ( Form PCT/ ISA / 237 ) issued on Dec . 1 , 2015 , in the International Application No . PCT / (73 ) Assignee : West Virginia University , US15 / 35784 . ( 19 pages ) . Morgantown , WV (US ) Š?igelová et al ., “ Biotransformation of Ergot Alkaloids by Plant Cell Cultures with High Peroxidase Activity, ” Biotechnology Let ( * ) Notice : Subject to any disclaimer , the term of this ters , (Nov . 1995 ) , vol. 17 , No . 11, pp . 1213 - 1218 . patent is extended or adjusted under 35 Singer et al. , “ High - Throughput TAIL -PCR as a Tool to Identify U . S . C . 154 ( b ) by 0 days . DNA Flanking Insertions, ” Methods in Molecular Biology, ( 2003 ) , vol. 236 , pp . 241 - 272 . ( 21 ) Appl. No. : 14 /739 , 382 ( Continued ) Î( 22 ) Filed : Jun . 15 , 2015 Primary Examiner - Robert Mondesi (65 ) Prior Publication Data Assistant Examiner — Richard Ekstrom US 2015 /0361471 A1 Dec . 17 , 2015 (74 ) Attorney , Agent, or Firm — Buchanan Ingersoll & Related U .S . Application Data Rooney PC ( 60 ) Provisional application No . 62/ 012 ,658 , filed on Jun . 16 , 2014 . ( 57 ) ABSTRACT (51 ) Int. Ci. C12P 17 / 18 ( 2006 .01 ) The present invention provides a method of producing C12N 15 /52 ( 2006 . 01 ) lysergic acid and other ergot alkaloids by genetic modifica (52 ) U . S . CI. tion of a fungus . A strain of fungus comprising Aspergillus CPC . .. .. C12P 17/ 182 ( 2013 .01 ); C12N 15 / 52 fumigatus ( A . fumigatus ) and expressing one or more genes ( 2013 .01 ) of the ergot alkaloid biosynthesis pathway from one or more ( 58 ) Field of Classification Search fungus selected from the group consisting of Epichloë None festucae var . loliixEpichloë typhina isolate Lpl ( E . sp . Lpl ) ; See application file for complete search history . Claviceps species ; Claviceps africana ( C . africana ); Clavi (56 ) References Cited ceps gigantea ( C . gigantea ); Epichloë coenophiala and Periglandula species, wherein gene easA or gene easM is U . S . PATENT DOCUMENTS inactivated in said A . fumigatus , is provided . 3 ,314 , 961 A 4 / 1967 Rutschmann et al. 2015 /0211036 A1 * 7 / 2015 Naesby .. .. C12N 9 / 001 435 / 122 18 Claims, 14 Drawing Sheets " ! NON W 5959 U IL TRAIN waste 1 - 2 , W ixxis 14: 29 - 12. 2 . 22 sigrics ? 233 tari ASK tunnelse: HIMSIAIN sok cic fumigadbazir: Nyergic rets Amer . 4492 EISA Muscypodium : US 9 ,790 , 528 B2 Page 2 ( 56 ) References Cited Ge et al. , “ Bioactive Alkaloids from Endophytic Aspergillus Fumigatus, ” Journal of Natural Products , ( 2009 ) , vol. 72 , No . 4 , pp . 753 -755 . OTHER PUBLICATIONS Goetz et al. , “ Ergot Cluster -Encoded Catalase is Required for Synthesis of Chanoclavine - I in Aspergillus Fumigatus, ” Current Tooley et al. , " Analysis of Claviceps Africana and C . Sorghi from Genetics , ( Jun . 2011 ) , vol. 57 , No . 3 , pp . 201 - 2011 . 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