Synthetic biology approach to synthesize nicotinic acid from 3-picoline

A method for the synthesis of nicotinic acid from 3-picoline, the method consisting of steps

• a. Extracting genes “C” “M”, “A” and “B”, distinctly from any of the genomes of selected organisms such as Pseudomonas putida, Arthrobacter woluwensis, Acidovorax sp., Acinetobacter calcoaceticus, Burkholderia sp., Croceicoccus sp., Cupriavidus sp., Delftia sp., Devosia sp., Geodermatophilus sp., Jatrophihabitans sp., Kribella sp., Lacisediminimonas sp., Microbacterium sp., Mycolicibacterium sp., Nocardioides sp., Novosphingobium sp., Parapusillimonas sp., Planosporangium sp., Prauserella sp., Ramlibacter sp., Rhodococcus sp wherein, the genes “C”, “M”, “A” and “B” encode the proteins benzaldehyde dehydrogenase, monooxygenase, electron transfer component of the monooxygenase and benzyl alcohol dehydrogenase, respectively and the genes “C”, “M”, “A” and “B” include the associated genetic components such as RBS and spacers from the respective genome
• b. Designing synthetic gene constructs with the extracted genes in the wild or engineered form, cloning the synthetic gene construct into an expression vector at specific restriction enzyme sites such as NcoI, NdeI, BamHI, EcoRI, HindIII, XhoI, and NotI;
• expressing these cloned genes within a transforming recombinant host cells, wherein the engineering of the genes are done involving site-directed mutagenesis, rational design, directed evolution, or a combination thereof.
• c. Culturing the transformed host cells under conditions suitable for expression of said genes to convert 3-picoline to nicotinic acid via enzymatic action of expressed proteins from said genes.