Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 18 July 1997:
Vol. 277. no. 5324, pp. 367 - 369
DOI: 10.1126/science.277.5324.367
Prev | Table of Contents | Next

Reports

Precursor-Directed Biosynthesis of Erythromycin Analogs by an Engineered Polyketide Synthase

John R. Jacobsen, C. Richard Hutchinson, David E. Cane, Chaitan Khosla *

A genetic block was introduced in the first condensation step of the polyketide biosynthetic pathway that leads to the formation of 6-deoxyerythronolide B (6-dEB), the macrocyclic precursor of erythromycin. Exogenous addition of designed synthetic molecules to small-scale cultures of this null mutant resulted in highly selective multimilligram production of unnatural polyketides, including aromatic and ring-expanded variants of 6-dEB. Unexpected incorporation patterns were observed, illustrating the catalytic versatility of modular polyketide synthases. Further processing of some of these scaffolds by postpolyketide enzymes of the erythromycin pathway resulted in the generation of novel antibacterials with in vitro potency comparable to that of their natural counterparts.

J. R. Jacobsen, Department of Chemical Engineering, Stanford University, Stanford, CA 94305-5025, USA.
C. R. Hutchinson, School of Pharmacy and Department of Bacteriology, University of Wisconsin, Madison, WI 53706, USA.
D. E. Cane, Department of Chemistry, Box H, Brown University, Providence, RI 02912, USA.
C. Khosla, Departments of Chemical Engineering, Chemistry, and Biochemistry, Stanford University, Stanford, CA 94305-5025, USA.
*   To whom correspondence should be addressed.

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Chain Initiation in the Leinamycin-producing Hybrid Nonribosomal Peptide/Polyketide Synthetase from Streptomyces atroolivaceus S-140: DISCRETE, MONOFUNCTIONAL ADENYLATION ENZYME AND PEPTIDYL CARRIER PROTEIN THAT DIRECTLY LOAD D-ALANINE.
G.-L. Tang, Y.-Q. Cheng, and B. Shen (2007)
J. Biol. Chem. 282, 20273-20282
   Abstract »    Full Text »    PDF »
Efficient Synthesis of Simvastatin by Use of Whole-Cell Biocatalysis.
X. Xie and Y. Tang (2007)
Appl. Envir. Microbiol. 73, 2054-2060
   Abstract »    Full Text »    PDF »
Characterization of a Large, Stable, High-Copy-Number Streptomyces Plasmid That Requires Stability and Transfer Functions for Heterologous Polyketide Overproduction.
R. Fong, Z. Hu, C. R. Hutchinson, J. Huang, S. Cohen, and C. Kao (2007)
Appl. Envir. Microbiol. 73, 1296-1307
   Abstract »    Full Text »    PDF »
Chemobiosynthesis of Novel 6-Deoxyerythronolide B Analogues by Mutation of the Loading Module of 6-Deoxyerythronolide B Synthase 1.
S. Murli, K. S. MacMillan, Z. Hu, G. W. Ashley, S. D. Dong, J. T. Kealey, C. D. Reeves, and J. Kennedy (2005)
Appl. Envir. Microbiol. 71, 4503-4509
   Abstract »    Full Text »    PDF »
Production of the Potent Antibacterial Polyketide Erythromycin C in Escherichia coli.
S. Peiru, H. G. Menzella, E. Rodriguez, J. Carney, and H. Gramajo (2005)
Appl. Envir. Microbiol. 71, 2539-2547
   Abstract »    Full Text »    PDF »
A specific role of the Saccharopolyspora erythraea thioesterase II gene in the function of modular polyketide synthases.
Z. Hu, B. A. Pfeifer, E. Chao, S. Murli, J. Kealey, J. R. Carney, G. Ashley, C. Khosla, and C. R. Hutchinson (2003)
Microbiology 149, 2213-2225
   Abstract »    Full Text »    PDF »
Biosynthetic Gene Cluster of Simocyclinone, a Natural Multihybrid Antibiotic.
A. Trefzer, S. Pelzer, J. Schimana, S. Stockert, C. Bihlmaier, H.-P. Fiedler, K. Welzel, A. Vente, and A. Bechthold (2002)
Antimicrob. Agents Chemother. 46, 1174-1182
   Abstract »    Full Text »    PDF »
Crystal structure of the macrocycle-forming thioesterase domain of the erythromycin polyketide synthase: Versatility from a unique substrate channel.
S.-C. Tsai, L. J. W. Miercke, J. Krucinski, R. Gokhale, J. C.-H. Chen, P. G. Foster, D. E. Cane, C. Khosla, and R. M. Stroud (2001)
PNAS 98, 14808-14813
   Abstract »    Full Text »    PDF »
Identification of the Novobiocin Biosynthetic Gene Cluster of Streptomyces spheroides NCIB 11891.
M. Steffensky, A. Mühlenweg, Z.-X. Wang, S.-M. Li, and L. Heide (2000)
Antimicrob. Agents Chemother. 44, 1214-1222
   Abstract »    Full Text »
A multiplasmid approach to preparing large libraries of polyketides.
Q. Xue, G. Ashley, C. R. Hutchinson, and D. V. Santi (1999)
PNAS 96, 11740-11745
   Abstract »    Full Text »    PDF »
Kinetic Analysis of the Actinorhodin Aromatic Polyketide Synthase.
J. Dreier, A. N. Shah, and C. Khosla (1999)
J. Biol. Chem. 274, 25108-25112
   Abstract »    Full Text »    PDF »
Microbial polyketide synthases: More and more prolific.
C. R. Hutchinson (1999)
PNAS 96, 3336-3338
   Full Text »    PDF »
A gene cluster for macrolide antibiotic biosynthesis in Streptomyces venezuelae: Architecture of metabolic diversity.
Y. Xue, L. Zhao, H.-w. Liu, and D. H. Sherman (1998)
PNAS 95, 12111-12116
   Abstract »    Full Text »    PDF »
Harnessing the Biosynthetic Code: Combinations, Permutations, and Mutations.
D. E. Cane, C. T. Walsh, and C. Khosla (1998)
Science 282, 63-68
   Abstract »    Full Text »
beta -Lactam synthetase: A new biosynthetic enzyme.
B. O. Bachmann, R. Li, and C. A. Townsend (1998)
PNAS 95, 9082-9086
   Abstract »    Full Text »    PDF »
Engineering Broader Specificity into an Antibiotic-Producing Polyketide Synthase.
A. F. Marsden, B. Wilkinson, J. Cortés, N. J. Dunster, J. Staunton, and P. F. Leadlay (1998)
Science 279, 199-202
   Abstract »    Full Text »
An A245T Mutation Conveys on Cytochrome P450eryF the Ability to Oxidize Alternative Substrates.
H. Xiang, R. A. Tschirret-Guth, and P. R. Ortiz de Montellano (2000)
J. Biol. Chem. 275, 35999-36006
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)