{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,28]],"date-time":"2026-06-28T10:44:10Z","timestamp":1782643450219,"version":"3.54.5"},"reference-count":41,"publisher":"Microbiology Society","issue":"8","funder":[{"name":"NSF","award":["0545860"],"award-info":[{"award-number":["0545860"]}]},{"name":"NSF","award":["1033070"],"award-info":[{"award-number":["1033070"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2011,8,1]]},"abstract":"<jats:p>Tetracyclines are clinically important aromatic polyketides whose biosynthesis is catalysed by bacterial type II polyketide synthases (PKSs). Tetracyclines are biosynthesized starting with an amide-containing malonamate starter unit and the resulting C-2 carboxyamide is critical for the antibiotic activities. In this work, we genetically verified that an amidotransferase, OxyD, and a thiolase, OxyP, are involved in the biosynthesis and incorporation of the starter unit. First, two mutations, R248T and D268N, were found to be present in OxyD* encoded in <jats:italic>Streptomyces rimosus<\/jats:italic> ATCC 13224, a strain that produces the acetate-primed 2-acetyl-2-decarboxyamido-oxytetracycline (ADOTC) instead of the malonamate-primed oxytetracycline (OTC). Homology modelling suggested that in particular D268N may inactivate OxyD. Complementation of <jats:italic>S. rimosus<\/jats:italic> ATCC 13224 with wild-type OxyD restored OTC biosynthesis, thereby confirming the essential role of OxyD in the synthesis of the amide starter unit. Second, using a series of knockout and complementation approaches, we demonstrated that OxyP is most likely involved in maintaining fidelity of the amide-priming process via hydrolysis of the competing acetate priming starter units. While the inactivation of OxyP does not eliminate OTC biosynthesis, the ratio of acetate-primed ADOTC to malonamate-primed OTC is significantly increased. This suggests that OxyP plays an ancillary role in OTC biosynthesis and is important for minimizing the levels of ADOTC, a shunt product that has much weaker antibiotic activities than OTC.<\/jats:p>","DOI":"10.1099\/mic.0.048439-0","type":"journal-article","created":{"date-parts":[[2011,5,27]],"date-time":"2011-05-27T02:36:09Z","timestamp":1306463769000},"page":"2401-2409","source":"Crossref","is-referenced-by-count":19,"title":["Genetic characterization of enzymes involved in the priming steps of oxytetracycline biosynthesis in Streptomyces rimosus"],"prefix":"10.1099","volume":"157","author":[{"given":"Peng","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, USA"},{"name":"Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200030, 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