Metabolic engineering of Pediococcus acidilactici BD16 for production of vanillin through ferulic acid catabolic pathway and process optimization using response surface methodology

• Published in: Applied microbiology and biotechnology 2014-10, Vol.98 (20), p.8539-8551
• Main Authors: Kaur, Baljinder, Chakraborty, Debkumar, Kumar, Balvir
• Format: Article
• Language: English
• Subjects: Acids; Actinobacteria - enzymology; Amycolatopsis; Bacteria; Benzaldehydes - metabolism; biochemical pathways; Biomass; Biomedical and Life Sciences; Biosynthesis; Biotechnological Products and Process Engineering; Biotechnology; Biotransformation; Coenzyme A Ligases - genetics; Coenzyme A Ligases - metabolism; Coumaric Acids - metabolism; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; E coli; Enoyl-CoA Hydratase - genetics; Enoyl-CoA Hydratase - metabolism; Enzymes; Escherichia coli; ferulic acid; Gene Expression; Genes; Genetic aspects; Genetic engineering; Gram-positive bacteria; GRAS substances; lactic acid bacteria; Life Sciences; Metabolic Engineering; Metabolic Networks and Pathways - genetics; Metabolism; Microbial Genetics and Genomics; Microbiology; Molecular Sequence Data; Natural products; Optimization; Pediococcus - genetics; Pediococcus - metabolism; Pediococcus acidilactici; Phenols; Physiological aspects; Pseudomonas; Pseudomonas - enzymology; Pseudomonas fluorescens; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; response surface methodology; Sequence Analysis, DNA; Spectral analysis; Streptomyces; Studies; Vanillin
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-014-5950-x

Occurrence of feruloyl-CoA synthetase (fcs) and enoyl-CoA hydratase (ech) genes responsible for the bioconversion of ferulic acid to vanillin have been reported and characterized from Amycolatopsis sp., Streptomyces sp., and Pseudomonas sp. Attempts have been made to express these genes in Escherichia coli DH5α, E. coli JM109, and Pseudomonas fluorescens. However, none of the lactic acid bacteria strain having GRAS status was previously proposed for heterologous expression of fcs and ech genes for production of vanillin through biotechnological process. Present study reports heterologous expression of vanillin synthetic gene cassette bearing fcs and ech genes in a dairy isolate Pediococcus acidilactici BD16. After metabolic engineering, statistical optimization of process parameters that influence ferulic acid to vanillin biotransformation in the recombinant strain was carried out using central composite design of response surface methodology. After scale-up of the process, 3.14 mM vanillin was recovered from 1.08 mM ferulic acid per milligram of recombinant cell biomass within 20 min of biotransformation. From LCMS-ESI spectral analysis, a metabolic pathway of phenolic biotransformations was predicted in the recombinant P. acidilactici BD16 (fcs ⁺/ech ⁺).