Eli Lilly and Company

Gene M Wild

Eugene T Seno

Richard H Baltz

Herbert A Kirst

Charles L Hershberger

Kurt E Merkel

Robert E Weeks

Robert L Hamill

Karen L Cox

Mack H McCormick

The present invention discloses a method for producing a novel antibiotic, 2''-O-demethyltylosin. The novel antibiotic, utilizing recombinant DNA technology, can be produced from a transformed mutant of a tylosin producing microorganism. By transforming for example Streptomyces fradiae GS 16 with plasmid pHJL284, the transformant can produce 2''-O-demethyltylosin. Streytomyces fradiae GS16 is a tylosin producing species that contain a mutation in the tylE gene found in the tylosin biosynthetic pathway. The tylE gene codes for demethylmacrocin 2''-O-methyltransferase enzyme (DMOMT), the enzyme that methylates the 2-hydroxyl position of the 6-deoxyallose moiety. Plasmid pHJL284 contains the cloned tylF gene which codes for the macrocin 3''-O-methyltransferase enzxyme (MOMT), the enzyme that methylates the 3''-hydroxyl position, but it does not contain the tylE gene. Transformation of S. fradiae GS16 with plasmid pHJL284 results in increased copies of the tylF gene in the absence of the tylE gene, therefore allowing an increased rate of methylation of the 3''-hydroxyl position in the absence of the methyl group at the 2''-hydroxyl position. The resulting transformant, therefore, can produce 2''-O-demethyltylosin in appreciable quantities. The present invention is not limited to S. fradiae.For example, two other tylosin producing species such as Streptomyces rimosus and Streptomyces hygroscopicus, may also be used for purposes of the invention disclosed. 2''-O-Demethyltylosin is useful as an antibiotic with a microbial inhibitory activity similar to tylosin.

Method for producing 2''-O-demethyltylosin

Glycopeptide antibiotic A-4696G is produced by culturing Actinoplanes missouriensis mutant strain ATCC 31681 under submerged aerobic fermentation conditions, and is recovered from the fermentation broth by resin adsorption and chromatographic purification. A-4696G inhibits the growth of bacteria pathogenic to man and animals and also enhances the growth of ruminants by increasing feed utilization.

Antibiotic A-4696 factor G

23-Demycinosyltylosin (DMT) which has the formula: ##STR1## 20-dihydro-DMT, specified acyl ester derivatives, and their acid addition salts are useful antibacterial agents. Improved methods of making 5-O-mycaminosyltylonolide (OMT) and 20-dihydro-OMT by mild acid hydrolysis of DMT and 20-dihydro-DMT, respectively, are included.

Process for the production of mycaminosyltylonolide

20-Dihydro-20-deoxy-23-de(mycinosyloxy)tylosin (20-deoxo-DMOT), specified 2'-acyl ester derivatives, and their acid addition salts are useful intermediates and antibacterial agents. Methods of preparing 20-deoxo-DMOT and 5-O-mycaminosyltylactone by fermentation of Streptomyces fradiae ATCC 31733 are included.

20-Dihydro-20-deoxy-23-de(mycinosyloxy)tylosin

2''-O-demethylmacrocin (DOMM) which has the formula: ##STR1## 20-dihydro-DOMM, 2''-O-demethyllactenocin (DOML), 20-dihydro-DOML, specified acyl ester derivatives, and their acid addition salts are useful antibacterial agents.

Demethylmacrocin compounds and derivatives thereof

Purification of glycopeptide antibiotics actaplanin and vancomycin by adsorption on non-functional resins followed by elution of the antibiotic from the resin with aqueous solvents.

Purification of glycopeptide antibiotics using non-functional resins

Process for producing 20-dihydro-20-deoxy-23-de(mycinosyloxy)tylosin

20-Dihydro-20-deoxy-23-demycinosyltylosin (DH-DO-DMT), 20-dihydro-20-deoxy-5-O-mycaminosyltylonolide (DH-DO-OMT), specified acyl ester derivatives, and their acid addition salts are useful intermediates and antibacterial agents. Methods of preparing DH-DO-DMT and DH-DO-OMT by fermentation of Streptomyces fradiae and the microorganism S. fradiae ATCC 31733 are included.

20-Dihydro-20-deoxy-23-demycinosyltylosin and process for its production

Fermentation process for producing demycinosyltylosin

Demethylmacrocin and process for its production

23-De(mycinosyloxy)tylosin (DMOT) which has the formula: ##STR1## 20-dihydro-DMOT, specified acyl ester derivatives, and the acid addition salts thereof are useful antibacterial agents. New methods of making 23-deoxy-5-O-mycaminosyltylonolide (DOMT) and 20-dihydro-DOMT by mild acid hydrolysis of DMOT and 20-dihydro-DMOT, respectively, are included.

Process for producing de(mycinosyloxy)tylosin

23-De(mycinosyloxy)tylosin (DMOT) which has the formula: ##STR1## 20-dihydro-DMOT, specified acyl ester derivatives, and the acid addition salts thereof are useful antibacterial agents. New methods of making 23-deoxy-5-O-my-caminosyltylonolide (DOMT) and 20-dihydro-DOMT by mild acid hydrolysis of DMOT and 20-dihydro-DMOT, respectively, are included.

Growing community of inventors

Indianapolis, IN, United States of America

Gene M Wild