Abstract
The current research study deals with the screening of a potent vanillin-producing microorganism among 96 isolated strains. Biochemical characterization and molecular identification confirmed that the isolated strain belongs to the Klebsiella pneumoniae bacteria, so it was denoted as Klebsiella pneumoniae P27. The optimization of medium components for the enhanced production of vanillin was carried out using two-stage statistical experimental designs, in which the significant medium components for vanillin production were screened using a Plackett-Burman experimental design. And the optimal levels of those noteworthy factors were determined by using central composite design. The statistical optimization of medium components resulted in increases in vanillin production and vanillyl alcohol oxidase activity of 2.05-fold and 3.055-fold, respectively. The highest vanillin production (30.88 mg/L) and vanillyl alcohol oxidase activity (0.044 U/mL) was observed after 16 h of incubation in the presence of 0.26 mL/L creosol, 8.06 g/L yeast extract and 2.77 g/L NH4NO3 in the production medium. The optimally produced vanillin was extracted and confirmed using FTIR and LCMS spectral analysis. The results of the current study support a statistical process optimization approach as a potential technique for the enhanced production of vanillin from creosol by using newly isolated Klebsiella pneumoniae P27 bacterial strain.
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