ATBECHDEDKESFRGBGRITLILUNLSEMCPTIESILTLVFIROMKCYALTRBGCZEEHUPLSK..HR............JDIM360 (Ver 1.5 26 Jan 2004) - 1999001/01477563CORRECTED EUROPEAN PATENT APPLICATIONA9A320050921W10000000071deBibliographieenBibliographyfrBibliographieEP04011223.7200405122004120220050620enenen471256 P20030516US2005092120053820041117200447200509212005387 7C 12N 15/31 A 7C 12N 15/53 B 7C 12N 15/54 B 7C 12N 15/55 B 7C 12N 1/21 B 7C 12N 9/16 B 7C 12N 9/02 B 7C 12N 9/10 B 7C 07K 14/36 B 7C 12P 17/18 BdeKlonierung von Streptomyces cyaneogriseus subsp.noncyanogenus Genen zur Biosynthese von Antibiotika und Verfahren zu ihrer VerwendungenCloning genes from streptomyces cyaneogriseus subsp.noncyanogenus for biosynthesis of antibiotics and methods of usefrClonage de gènes de Streptomyces cyaneogriseus subsp.noncyanogenus pour la biosynthèse des antibiotiques et procédés pour leur utilisationWyeth04088651AM-100484Five Giralda FarmsMadison, New Jersey 07940USHuang, Chengjin1169 26th Avenue NorthFort Dodge, IA 50501USChaleff, Deborah Tardy31 Arvida DrivePennington, NJ 08534USRuppen, Mark Edward6 Lea CourtGarnerville, NY 10923USStephens, Jerome21595 Scenic DriveMentone, AL 35984USIngham, Stephen H.et al00128401Wyeth Pharmaceuticals, Patents & Trade Marks Dept., Huntercombe Lane South, TaplowMaidenhead, Berkshire SL6 0PHGBDeclaration under Rule 28(4) EPC (expert solution)ATBEBGCHCYCZDEDKEEESFIFRGBGRHUIEITLILUMCNLPLPTROSESISKTRAL20040512LT20040512LV20040512MK2004051220041124200448

The present invention relates to the complete biosynthetic pathway for the formation of the LL-F28249 compounds and, most importantly, the major component LL-F28249α. The purified and isolated nucleic acid molecule encoding the proteins of the biosynthetic pathway, which is isolated from a wild-type or mutant Streptomyces, is fully described in SEQ ID NO:1. The DNA gene cluster and its expression in a suitable host enable the efficient production of the highly active natural metabolites and semisynthetic derivatives. The invention further concerns plasmids, vectors and host cells that contain and express the novel nucleic acid molecule. Of particular interest, the entire biosynthetic pathway fits compactly in three plasmids, Cos11, Cos36 and Cos40. The invention also concerns the purified and isolated biosynthesis proteins that are encoded by the whole DNA gene cluster. Additionally, the invention involves a new efficient, biochemical method of preparing moxidectin.