Science and Technology Production
Abstract book - Production of a non-natural chemicals by MenD from E. coli
Congreso
Date:
2014Publishing House and Editing Place:
Thiamine CongressSummary *
Traditional chemistry is often based on rare and limited resources and produces frequently hazardous waste. [1] Alternatively, formation of fine chemicals by biotechnological processes overcomes these negative aspects but it is restricted to natural pathways. [2] Producing non- natural chemicals using microorganisms is still a challenge.In this work, metabolic pathway engineering was combined with the ability of the E. coli enzyme MenD to produce fine chemicals by whole-cell biotransformation. MenD catalyzes a Stetter-like 1,4-addition of α-ketoglutarate (α-KG) and isochorismate (1) leading to 2- succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylic acid (SEPHCHC, 2), an intermediate in the menaquinone biosynthesis. [3].MenD is also able to transform the non-physiological substrate 5,6-dihydroxycyclohexa-1,3- dienecarboxylic acid (2,3-CHD, 3). [4] The enzyme was functionally expressed in genetically engineered E. coli strains that are 2,3- CHD overproducers. The non-natural compound 2-succinyl-5,6-dihydroxycyclohex-3-ene-1- carboxylic acid (SDHCHC, 4) was then obtained in 50 mg/L by fermentation processes in shaking flasks. By means of culture conditions optimization, changing the producing strain and rational design of the active site, the yield was further improved to 2.5 g SDHCHC per liter culture medium. Information provided by the agent in SIGEVAKey Words
THIAMINE-DEPENDENT ENZYMESMETABOLIC ENGINEERING