The wild Saccharomyces Cerevisiae consider as a good candidate for synthesizing high quantity of glutathione (red). GSH1 gene from Saccharomyces cerevisiae was identified and cloned in E.coli expression vector for over expression of glutathione synthetase leads to increase the levels of GSH in cellular extracts and the product purified to confirm its expression. Primers were designed based on entire sequence of Saccharomyces cerevisiae and GC content of the genomic DNA. The PCR carried out to amplify the template DNA with the primers. The amplified products were inserted in 1-67 & 99-346 region of PUC18 used as a cloning vector for the higher expression of glutathione production. In view of comparative analysis of glutathione preparation and standarized the protocol used in various steps involved in the purification process, subsequently identify the product recovered from different samples used in the experiment. The cellular glutathione content extracted in a supernatant of the culture lysate used for determined of glutathione. The efficiency of GSH production by recombinant E.coli has nearly 40 fold increased its production when compared to wild type strains. The prepared fractions were used for analysis of glutathione also the size of final purified products were checked by SDS PAGE and concentrated by lyophilization process, it is recommended that application of glutathione on protecting freeze dried molecules in various field.
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