Vanillin is undoubtedly one of the most popular and widely used flavoring agents in the world. Natural vanillin is extracted from vanilla beans and is relatively expensive. Moreover, the consumer demand for natural vanillin highly exceeds the amount of vanillin extracted by plant sources. This had led to the investigation of alternative routes for its production such as microbial bioconversion. In this study, a novel strain bacterium capable of converting isoeugenol to vanillin was isolated by conventional enrichment process from soils of spicy plants farms. On the basis of morphological and physiochemical characteristics, the isolate was identified as Pseudomonas sp. strain ISPC2. Vanillin formation was analyzed by spectrophotometery with thiobarbituric acid reagent and evaluated accurately by gas chromatography. Using 10 g/l of isoeugenol as substrate in 25 ml reaction solution at 30 °C and 150 rpm, vanillin reached a maximum concentration of 1.15 g/l after 96 h reaction, corresponding to a molar yield of 12.4%. This strain showed potential to be a good candidate for biotechnological production of vanillin from isoeugenol. Further studies for standardization and optimization for higher yield of vanillin production needs to be investigated.