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Diversity oriented biosynthesis via accelerated evolution of modular gene clusters.
Wlodek, Aleksandra; Kendrew, Steve G; Coates, Nigel J; Hold, Adam; Pogwizd, Joanna; Rudder, Steven; Sheehan, Lesley S; Higginbotham, Sarah J; Stanley-Smith, Anna E; Warneck, Tony; Nur-E-Alam, Mohammad; Radzom, Markus; Martin, Christine J; Overvoorde, Lois; Samborskyy, Markiyan; Alt, Silke; Heine, Daniel; Carter, Guy T; Graziani, Edmund I; Koehn, Frank E; McDonald, Leonard; Alanine, Alexander; Rodríguez Sarmiento, Rosa María; Chao, Suzan Keen; Ratni, Hasane; Steward, Lucinda; Norville, Isobel H; Sarkar-Tyson, Mitali; Moss, Steven J; Leadlay, Peter F; Wilkinson, Barrie; Gregory, Matthew A.
Afiliación
  • Wlodek A; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Kendrew SG; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Coates NJ; Engineered Biodesign Limited, Cambridge, CB22 3GN, UK.
  • Hold A; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Pogwizd J; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Rudder S; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Sheehan LS; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Higginbotham SJ; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Stanley-Smith AE; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Warneck T; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Nur-E-Alam M; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Radzom M; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Martin CJ; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Overvoorde L; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Samborskyy M; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Alt S; Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, 12372, Saudi Arabia.
  • Heine D; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Carter GT; BASF SE, Speyerer Str. 2, Limburgerhof, 67117, Germany.
  • Graziani EI; Biotica Technology Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • Koehn FE; Isomerase Therapeutics Ltd., Chesterford Research Park, Cambridge, CB10 1XL, UK.
  • McDonald L; Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, UK.
  • Alanine A; Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK.
  • Rodríguez Sarmiento RM; Department of Molecular Microbiology, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK.
  • Chao SK; Chemical and Screening Sciences, Wyeth Pharmaceuticals, 401 North Middletown Road, Pearl River, NY, 10965, USA.
  • Ratni H; Chemical and Screening Sciences, Wyeth Pharmaceuticals, 401 North Middletown Road, Pearl River, NY, 10965, USA.
  • Steward L; Medicine Discovery Network-Synthetic Biology, Pfizer Worldwide R&D, 445 Eastern Point Rd., Groton, CT, 06340, USA.
  • Norville IH; Chemical and Screening Sciences, Wyeth Pharmaceuticals, 401 North Middletown Road, Pearl River, NY, 10965, USA.
  • Sarkar-Tyson M; Chemical and Screening Sciences, Wyeth Pharmaceuticals, 401 North Middletown Road, Pearl River, NY, 10965, USA.
  • Moss SJ; Roche Innovation Center Basel, Pharmaceutical Research and Early Development (PRED), Basel, CH-4070, Switzerland.
  • Leadlay PF; Roche Innovation Center Basel, Pharmaceutical Research and Early Development (PRED), Basel, CH-4070, Switzerland.
  • Wilkinson B; Roche Innovation Center Basel, Pharmaceutical Research and Early Development (PRED), Basel, CH-4070, Switzerland.
  • Gregory MA; Roche Innovation Center Basel, Pharmaceutical Research and Early Development (PRED), Basel, CH-4070, Switzerland.
Nat Commun ; 8(1): 1206, 2017 10 31.
Article en En | MEDLINE | ID: mdl-29089518
Erythromycin, avermectin and rapamycin are clinically useful polyketide natural products produced on modular polyketide synthase multienzymes by an assembly-line process in which each module of enzymes in turn specifies attachment of a particular chemical unit. Although polyketide synthase encoding genes have been successfully engineered to produce novel analogues, the process can be relatively slow, inefficient, and frequently low-yielding. We now describe a method for rapidly recombining polyketide synthase gene clusters to replace, add or remove modules that, with high frequency, generates diverse and highly productive assembly lines. The method is exemplified in the rapamycin biosynthetic gene cluster where, in a single experiment, multiple strains were isolated producing new members of a rapamycin-related family of polyketides. The process mimics, but significantly accelerates, a plausible mechanism of natural evolution for modular polyketide synthases. Detailed sequence analysis of the recombinant genes provides unique insight into the design principles for constructing useful synthetic assembly-line multienzymes.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Variación Genética / Familia de Multigenes / Evolución Molecular / Vías Biosintéticas Idioma: En Revista: Nat Commun Asunto de la revista: BIOLOGIA / CIENCIA Año: 2017 Tipo del documento: Article Pais de publicación: Reino Unido

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Variación Genética / Familia de Multigenes / Evolución Molecular / Vías Biosintéticas Idioma: En Revista: Nat Commun Asunto de la revista: BIOLOGIA / CIENCIA Año: 2017 Tipo del documento: Article Pais de publicación: Reino Unido