Identification of the Novobiocin Biosynthetic Gene Cluster of Streptomyces spheroides NCIB 11891

doi:10.1128/AAC.44.5.1214-1222.2000

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Antimicrobial Agents and Chemotherapy, May 2000, p. 1214-1222, Vol. 44, No. 5
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification of the Novobiocin Biosynthetic Gene Cluster of Streptomyces spheroides NCIB 11891

Marion Steffensky, Agnes Mühlenweg, Zhao-Xin Wang, Shu-Ming Li, and Lutz Heide^*

Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhard-Karls-Universität Tübingen, 72076 Tübingen, Germany

Received 12 August 1999/Returned for modification 9 December 1999/Accepted 29 January 2000

The novobiocin biosynthetic gene cluster from Streptomyces spheroides NCIB 11891 was cloned by using homologous deoxynucleosidediphosphate (dNDP)-glucose 4,6-dehydratase gene fragments as probes.Double-stranded sequencing of 25.6 kb revealed the presence of23 putative open reading frames (ORFs), including the gene fornovobiocin resistance, gyrB^r, and at least 11 further ORFs to which a possible role in novobiocinbiosynthesis could be assigned. An insertional inactivation experimentwith a dNDP-glucose 4,6-dehydratase fragment resulted in abolishmentof novobiocin production, since biosynthesis of the deoxysugarmoiety of novobiocin was blocked. Heterologous expression of akey enzyme of novobiocin biosynthesis, i.e., novobiocic acid synthetase,in Streptomyces lividans TK24 further confirmed the involvementof the analyzed genes in the biosynthesis of theantibiotic.

^* Corresponding author. Mailing address: Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhard-Karls-Universität Tübingen,Auf der Morgenstelle 8, 72076 Tübingen, Germany. Phone: 49-7071-29-72460.Fax: 49-7071-29-5250. E-mail: heide{at}uni-tuebingen.de.

Antimicrobial Agents and Chemotherapy, May 2000, p. 1214-1222, Vol. 44, No. 5
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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