Biosynthesis of Indigo Using Recombinant E. Coli: Development of a Biological System for the Cost Effective Production of a Large Volume Chemical
BIOSYNTHESIS OF INDIGO USING RECOMBINANT E. COLI: DEVELOPMENT OF A BIOLOGICAL SYSTEM FOR THE COST EFFECTIVE PRODUCTION OF A LARGE VOLUME CHEMICAL Alan Berry, Ph.D., Senior Scientist; Scott Battist, B.S., Ch.E., Development Engineer; Gopal Chotani, Ph.D., Ch.E., Senior Staff Scientist; Tim Dodge, M.S., Ch.E., Senior Scientist; Steve Peck, B.S., Ch.E., Development Engineer; Scott Power, Ph.D., Vice President and Research Fellow; and Walter Weyler, Ph.D., Senior Scientist Genencor International. Rochester, NY. 14618-3916. Abstract Cost-effective production of any large-volume chemical by fermentation requires extensive manipulation ofboth the production organism and the fermentation and recovery processes. We have developed a recombinant E. coli system for the production of tryptophan and several other products derived from the aromatic amino acid pathway. By linking our technology for low-cost production oftryptophan from glucose with the enzyme naphthalene dioxygenase (NOO), we have achieved an overall process for the production ofindigo dye from glucose. To successfully join these two technologies, both the tryptophan pathway and NOD were extensively modified via genetic engineering. In addition, systems were developed to remove deleterious by-products generated during the chemical oxidations leading to indigo formation. Low-cost fermentation processes were developed that utilized minimal-salts media containing glucose as the sole carbon source. Finally, economical recovery processes were used that preserved the environmental friendliness ofthe biosynthetic route to indigo. 1121 Introduction Two major problems facing the U.S. chemical industry are dependence on foreign petroleum and mounting environmental concerns and regulations (for review, see reference [1]). This has led to consideration ofbiosynthetic routes for production of aromatic compounds.
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