High-level expression and optimization of pantoate-β-alanine ligase in Bacillus megaterium for the enhanced biocatalytic production of D-pantothenic acid

D-Pantothenic acid (DPA), also known as vitamin B-5 is associated with several biological functions and its deficiency causes metabolic and energetic disorders in humans. Fortification of foods with DPA is the viable option to address this risk. DPA biological production route employs pantoate-beta-alanine ligase (PBL) as the key enzyme, which avoids the tedious and time-consuming optical resolution process. The selection of an efficient PBL enzyme is vital for the biological production of DPA. In this study, the panC gene encoding PBL from Escherichia coli, Bacillus megaterium, Corynebacterium glutamicum and Bacillus subtilis was expressed in B. megaterium. B. subtilis derived panC exhibited high PBL activity 61.62 +/- 2.15 U/mL. Co-expression of phosphoenolpyruvate carboxykinase (pckA) did not improve the DPA production in B. megaterium. Biocatalytic fed-batch fermentation with externally supplemented precursor substrates (D-pantoic acid and beta-alanine) improved DPA titer to 45.56 +/- 0.53 g/L. Daily dietary requirements of DPA for different age groups (including babies, small children, athletes and elderly people) is steadily increasing and the improved DPA production addressed in this study offers advantage for its application in fortification of food products meeting the emerging nutritional demand.

Automated summary

D-Pantothenic acid (DPA), also known as vitamin B-5, plays a crucial role in metabolic and energetic balance, and fortifying foods with DPA is a viable solution. Study showed that PBL enzyme derived from Bacillus megaterium exhibited high activity, and external supplementation of precursor substrates during fermentation improved DPA production.