Application of crude natural enzymes for extraction of Wali seed [Brucea javanica (L) Merr]

Abstract

Bruce javanica (L) Merr is a medicinally important plant, commonly known as Wali. The extract of its seed has various secondary metabolites responsible for its pharmacological properties. Unfortunately, the recent method of extraction lacks in the percentage of the yield and has disadvantages to the environment. Enzymatic extraction of bioactive compounds is a potential alternative. In this research, we use some crude natural enzymes that are amylase, protease (bromelain), and cellulase isolated from corn kernels, pineapple fruit, and beef rumen fluid, respectively. The aim of this research was to determine the effectiveness of crude natural enzymes for the extraction of metabolite compounds in wali seed comparing with the solvent extraction method. For each of one gram of wali seed powder was extracted by using ultrasonic-Assisted extraction (UAE) method with solvent of (1) methanol; (2) ethanol 96%; (3) bromelain; (4) amylase; (5) cellulase; (6) bromelain+amylase; (7) bromelain+cellulase; (8) amylase+cellulase; (9) amylase+bromelain+cellulase. The extracts of (1) and (2) were then evaporated; while extracts of (3) to (9) were freeze-dried. The appearance of extracts (1) and (2) were bright-yellow viscous liquid; while extracts (3) to (9) were solid-milky powder. Cellulase increases the extraction yield up to 40% and 22% in comparison with methanol and ethanol, respectively. The semiquantitative analysis showed that methanol extracts the metabolites with the highest concentration but has the lowest amount of yield extract. However, cellulase showed the highest ability to extract metabolites tested. The flavonoid content extracted using cellulase enzymes resulted in an increase of 273% and 170% in comparison with methanol and ethanol, respectively. Thus, the use of crude cellulase enzyme isolated from beef rumen fluid is the best choice to extract wali seed among other enzymes and solvents used.