Identification of cultivation condition to produce correctly folded form of a malaria vaccine based on Plasmodium falciparum merozoite surface protein-1 in Escherichia coli.
The C-terminal, 19-kDa domain of Plasmodium falciparum merozoite surface protein-1 (PfMSP-1(19)) is among the leading vaccine candidate for malaria due to its essential role in erythrocyte invasion by the parasite. We designed a synthetic gene for optimal expression of recombinant PfMSP-1(19) in Escherichia coli and developed a scalable process to obtain high-quality PfMSP-1(19). The synthetic gene construct yielded a fourfold higher expression level of PfMSP-1(19) in comparison to the native gene construct. Optimization of cultivation conditions in the bioreactor indicated important role of yeast extract and substrate feeding strategy for obtaining enhanced expression of soluble and correctly folded PfMSP-1(19). It was observed that the higher expression level of PfMSP-1(19) was essentially associated with the generation of higher level of incorrectly folded PfMSP-1(19). A simple purification procedure comprising metal affinity and ion exchange chromatography was developed to purify correctly folded form of PfMSP-1(19) from cell lysate. Biochemical and biophysical characterization of purified PfMSP-1(19) suggested that it was highly pure, homogeneous, and correctly folded.
Mazumdar
Suman
S
Malaria Research Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), P.O. Box No 10504, Aruna Asaf Ali Marg, New Delhi, 110067, India.
Sachdeva
Suraksha
S
Chauhan
Virander S
VS
Yazdani
Syed Shams
SS
D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D017780:Malaria Vaccines / Q000096:biosynthesis*; D020066:Merozoite Surface Protein 1 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D010963:Plasmodium falciparum / Q000276:immunology*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D015003:Yeasts
D017780:Malaria Vaccines; D020066:Merozoite Surface Protein 1