This work proposed a study of direct enzymatic of vanillin formation by using rumen fluid which has enzymatic capability for tissue disruption of vanilla green pods to avoid the curing process. Application of enzymes during the formation of vanilla aromas and flavors and its extraction present nice opportunity to improve productivity, as the enzymatic reaction possibly substitute the microbial process in the traditional fermentation. Glucovanillin, the precursor of vanillin, contacted with the B-glucosidase in the green pods by destructing the cell wall. Liquid rument was providing enzyme for cell wall degradation. The contact of glucovanillin and B-glucosidase lead the hydrolysis of glucovanillin into vanillin. The amounts of glucovanillin and vanillin were examined by using HPLC analysis. The identification of vanillin was investigated by using liquid chromatography-mass spectrofotometry. Vanillin content of vanilla green pods was found higher in which by treating the vanilla green pods at 30°C.