聚合酶
DNA聚合酶
分子生物学
生物
重组DNA
聚合酶链反应
紫胶操纵子
酶
生物化学
基因
作者
Eva Agustriana,Isa Nuryana,Fina Amreta Laksmi,Kartika Sari Dewi,Hans Wijaya,Nanik Rahmani,Danu Risqi Yudiargo,Astadewi Ismadara,Helbert,Moch. Irfan Hadi,Awan Purnawan,Apridah Cameliawati Djohan
标识
DOI:10.1080/10826068.2022.2095573
摘要
Bst DNA polymerase is a DNA polymerase derived from Geobacillus stearothermophilus, has a strand-displacement activity, and is used in loop-mediated isothermal amplification (LAMP) for rapid detection of COVID-19. Despite its potential to be employed in the detection of COVID-19, using commercially available enzymes is not economically feasible. The use of noncommercial enzyme for routine use is desirable. However, research on Bst DNA polymerase is still limited in Indonesia. For those reasons, a preliminary study of scale-up production of recombinant Bst polymerase was conducted. Therefore, the optimization of expression conditions was performed. The optimum conditions for Bst polymerase expression were as follows: 1 mM of IPTG, post-induction incubation time of 6 h, and induction at OD600 1.1. Employing optimum conditions could result in 2.8 times increase in protein yield compared to the initial conditions. Subsequently, an operation in 1 L working volume by a lab-scale bioreactor had been performed, followed by purification and dialysis. The optimum result for a 1 L lab-scale bioreactor was achieved by applying 100 rpm and 3 vvm, giving 11.7 mg/L of protein yield. Bst polymerase was successfully purified showing 813.56 U/mg of polymerase activity.
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