| Abstrak/Abstract |
The large-scale production of Penicillin G Acylase (PGA) at high levels of soluble protein content is achievable through recombinant genetic techniques and expression within a specific host organism. Escherichia coli (E. coli) remains a popular choice as a bacterial host due to its rapid growth, costeffectiveness, and high expression rate. Despite its advantages, using E. coli as a production host presents disadvantages, notably in the proper folding of recombinant proteins, often resulting in biological inactivity. Various strategies have been developed to overcome these issues. These include selecting specific host strains (such as E. coli HB101 & JM109), utilizing fusion proteins to enhance the recovery of soluble proteins (e.g., MBP & NusA), optimizing fermentation conditions (e.g., low-temperature incubation), optimizing the protein isolation process for the recovery of active PGA (e.g., Freeze-thawing technique), and optimizing pH, temperature, and substrate specificity during the synthesis of β-lactam class antibiotics. This study proposes a solution to increase the expression of soluble PGA protein within biological hosts. By replacing the expression host and employing genetic engineering techniques, the study aims to achieve a high expression level of the PGA enzyme in an active form while simplifying the purification process |
