Studies on the production of shikimic acid using the aroK knockout strain of Bacillus megaterium.

Abstract

Shikimic acid has various pharmaceutical and industrial applications. It is the sole chemical building block for the antiviral drug oseltamivir (Tamiflu(®)) and one of the potent pharmaceutical intermediates with three chiral centres. Here we report a modified strain of Bacillus megaterium with aroK (shikimate kinase) knock out to block the aromatic biosynthetic pathway downstream of shikimic acid. Homologous recombination based gene disruption approach was used for generating aroK knock out mutant of B. megaterium. Shake flask cultivation showed shikimic acid yield of 2.98g/L which is ~6 times more than the wild type (0.53g/L). Furthermore, the shikimate kinase activity was assayed and it was 32% of the wild type. Effect of various carbon sources on the production of shikimic acid was studied and fructose (4%, w/v) was found to yield maximum shikimic acid (4.94g/L). The kinetics of growth and shikimic acid production by aroK knockout mutant was studied in 10L bioreactor and the yield of shikimic acid had increased to 6g/L which is ~12 fold higher over the wild type. It is evident from the results that aroK gene disruption had an immense effect in enhancing the shikimic acid production.