Team:Penn State/VanillinProject
From 2013.igem.org
Vanillin in Novel Plants Project
Vanillin is one of the major compounds in the vanilla flavor. A phenolic aldehyde, vanillin is also used in the pharmaceutical industry, beverages, as well as a fragrant compound in different products. The demand of Vanillin exceeded the natural production, so annually most vanillin is produced via chemical synthesis. We intend to produce vanillin in different plants as a natural alternative for vanillin production.
Introduction
Whether it’s a child’s first ice cream cone or the secret ingredient of a five star chef, everyone loves vanilla flavoring. Europeans first became aware of vanilla around 1520 and ever since then countless products featuring the beloved aromatic have been created. Vanillin, one of the major compounds in the vanilla flavor, is often produced by chemical synthesis due to the scarcity and high cost of natural vanilla extract. Hydroxycinnamoyl-CoA hydratase-lysase, more commonly known as HCHL, is an enzyme that converts feruloyl-CoA to vanillin and acetyl-CoA. HCHL is naturally produced by Pseudomonas putida Kt2440, a gram-negative rod-shaped saprotrophic soil bacterium. As plants naturally possess large pools of phenolic compounds used in lignin biosynthesis, we hope to be able to express HCHL within plants to convert feruoyl-CoA to vanillin.
Background
In order to express this enzyme within plants, HCHL must be cloned from Pseudomonas putida and modified for codon optimization in plants. This modification will involve the use of the Kozak sequence, which through experimental observation has been recognized as playing a major role in translational initiation. Additionally, a double stop codon along with the CaMV 35s promoter and Nos terminator will be utilized.
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