High level production of p-hydroxybenzoic acid in green plants

A method for the production of para-hydroxy benzoic acid in a green plant comprising:

• a) providing a green plant having an endogenous source of chorismate and containing a chorismate pyruvate lyase expression cassette having the following structure:
• P-T-C-D-CPL
• wherein:
• P is a promoter suitable for driving the expression of a chorismate pyruvate lyase gene;
• T is a nucleic acid molecule encoding a rubisco chloroplast transit peptide;
• C a nucleic acid molecule encoding a Rubisco chloroplast transit peptide cleavage site;
• D is a nucleic acid molecule encoding from about 4 to about 20 contiguous amino acids of the N-terminal portion of a Rubisco chloroplast transit peptide donor polypeptide;
• and
• CPL is a nucleic acid molecule encoding a mature chorismate pyruvate lyase protein;
• wherein each of P, T, C, D, and CPL are operably linked such that expression of the cassette results in translation of a chimeric protein comprising a chloroplast targeting sequence fused to the N-terminus of the mature chorismate pyruvate lyase protein;
• b) growing said plant under conditions whereby the chimeric protein is expressed and translocated to the chloroplast for the conversion of chorismate to para-hydroxy benzoic acid glucoside and para-hydroxy benzoic acid derivatives;
• c) recovering para-hydroxy benzoic acid and para-hydroxy benzoic acid derivatives from the plant;
• and
• d) processing said the para-hydroxy benzoic acid glucoside and para-hydroxy benzoic acid derivatives to free para-hydroxy benzoic acid.