Perpustakaan Digital ITB

The use of terpenoids is ubiquitous in several industries due to its diverse beneficial functions. As the demand of terpenoids increases, production of terpenoids should be able to compensate for it. Due to the low content of some terpenoids found in nature, alternative methods should be pursued to enhance the quantity of terpenoids, and in this study, genetic engineering of the plant synthetic pathway was chosen. A previous study was already conducted as an effort to construct the upper mevalonate pathway to increase the expression of mevalonate intermediate products in Escherichia coli. This study was performed to express the three key genes of the upper mevalonate pathway (atoB, mvaS, and mvaA) in Bacillus subtilis to produce mevalonate. The three upper mevalonate pathway genes have been successfully transformed in B. subtilis. Result for the expression was confirmed using SDS-PAGE and the mevalonate metabolites confirmation using GC-MS. From the SDS-PAGE result stained using coomassie blue dye, there were no apparent protein bands around the desired size of the target proteins (40.35 kDa for AtoB, 43.22 kDa for MvaS, and 46.18 kDa for MvaA). However, based on GC-MS measurement, mevalonate was present in the overexpression culture indicating the functionality and expression of the gene construct. Further optimization on the gene’s expression would be required to enhance mevalonate production in B. subtilis.