Dewi Mustikaningtyas, Aditya Marianti, Safira Chairani Dimarti, Shahnas Millenia Safitri, Maria Andini Oktaviana, Lailatul Faris Rosyidah
Glutathione is made up of the three essential amino acids cysteine, glutamic acid, and glycine. Promising prospects to enhance GSH production can be realized by applying genetic engineering with Saccharomyces cerevisiae. The method involves the strategic integration of the gsh1 and gex1 genes, achieved via the pESC-TRP plasmid insertion. This study aimed to accomplish two main objectives: firstly, the analysis of Gsh1 and Gex1 protein detection results; and secondly, the assessment of GSH1 enzyme activity in S. cerevisiae W303-1b. The employed methodologies encompassed two distinct approaches: (1) The SDS-PAGE method facilitated the qualitative analysis of Gsh1 and Gex1 proteins in transformant strains; (2) The ELISA reader method was employed to evaluate GSH1 enzyme activity in both transformant and wild-type strains. The outcomes exhibited distinct profiles: the Gsh1 protein featured a molecular weight of 58 kDa, while the Gex1 protein displayed a molecular weight of 68.9 kDa. Notably, the transformant strain stimulated by galactose induction exhibited the highest GSH1 enzyme activity, as evident from the glutathione production levels reaching 104.478 mg/L. This heightened glutathione yield in the transformant strain was attributed to the successful insertion of the gsh1 and gex1 genes, effectively realizing their overexpression mechanism within the cellular. © 2025 Author(s).
Biology Study Program, Faculty of Mathematics and Sciences, Universitas Negeri Semarang, Semarang, 50229, Indonesia; Public Health Study Program, Faculty of Medical, Universitas Negeri Semarang, Semarang, 50229, Indonesia
