(15c) Engineering of ADP1 for Production of Vanillin from Lignin

Vanillin is one of the most widely used natural flavoring agents in the world. Its popularity exceeds the supply from traditional plant extraction. Therefore, chemical synthesis has been used to meet global demand. Although this has successfully improved vanillin availability and pricing, growing consumer preferences for “natural” ingredients have steered food and agriculture firms to instead rely on biologically derived vanillin in their products. This forces the need for a creative solution for vanillin products to be both natural and affordable. Biotechnological approaches have been used to meet this need, with glucose and ferulic acid used as starting materials for a biological conversion to vanillin to date. Here, we look to harness the bacteria ADP1’s natural ability to degrade lignin monomers to produce vanillin from biomass. Our approach involves engineering ADP1 by deleting ADP1’s natural vanillic acid and protocatechuate degradation pathways and by introducing a catechol o-methyltransferase to convert protocatechuate to vanillic acid. In addition, we look to introduce a decarboxylase enzyme to convert vanillic acid to vanillin, to potentially provide an affordable and renewable source of natural vanillin.