@article{Alimuddin_Ali_Alim_Amin_2023, title={Enhancing Solubility of Recombinant Plasmodium Lactate Dehydrogenase (pLDH) Using Combination of Cold-Inducible Expression System and Cold-Stirred Bioreactor}, volume={30}, url={https://journal.ipb.ac.id/index.php/hayati/article/view/40340}, DOI={10.4308/hjb.30.3.561-566}, abstractNote={<p>A major drawback associated with an expression of a high-level Plasmodium Lactate Dehydrogenase (pLDH) using <em>Escherichia coli</em> is the low solubility due to the formation of an inclusion body (IB). This study aimed to develop a suitable protocol for enhancing the solubility of pLDH expressed in <em>E. coli</em>. Firstly, a pLDH-encoding gene was amplified from the blood sample of malaria-infected patients and ligated into pBlueScript II KS+ for sequencing. Afterward, the pLDH gene was digested and cloned into pColdTF for expression. The recombinant plasmid was transformed into the <em>E. coli</em> BL21 (DE3) RIPL Codon Plus Strain. Then, the bacterial host was initially cultured at 37°C until reaching optical density (OD) at 600 nm: 0.5. Thereafter, the growth temperature was lowered to 15°C, followed by the addition of 0.1 mM IPTG into the culture medium for inducing pLDH expression. Thereafter, the bacterial hosts were cultured in a cold-stirred bioreactor (15°C). The result showed that a combination of the low culture conditions (15°C) and a low amount of IPTG increased the solubility of pLDH. This result suggests that this protocol can be a convenient method for generating high-quality recombinant protein using the <em>E. coli</em> system.</p&gt;}, number={3}, journal={HAYATI Journal of Biosciences}, author={Alimuddin and Ali, Muhamad and Alim, Sahrul and Amin, Muhamad}, year={2023}, month={Mar.}, pages={561-566} }