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We utilized recombinant deoxyribonucleic acid (DNA) technology approaches to express soluble TbPLA<jats:sub>2<\/jats:sub> in <jats:italic>Escherichia coli<\/jats:italic> and <jats:italic>Pichia pastoris<\/jats:italic> and biochemically characterize the purified enzyme. Full\u2010length TbPLA<jats:sub>2<\/jats:sub> was insoluble and deposited as inclusion bodies when expressed in <jats:italic>E. coli<\/jats:italic>. However, soluble and active forms were obtained when both the full\u2010length and truncated TbPLA<jats:sub>2<\/jats:sub> were expressed in fusion with N\u2010terminal FLAG tag and C\u2010terminal eGFP in <jats:italic>P. pastoris<\/jats:italic>, and the truncated protein in fusion with N\u2010terminal FLAG tag and C\u2010terminal mClover in <jats:italic>E. coli<\/jats:italic>. Truncated TbPLA<jats:sub>2<\/jats:sub> lacking the signal peptide and transmembrane domain was finally expressed in Rosetta 2 cells and purified to homogeneity. Its migration on sodium dodecyl polyacrylamide gel electrophoresis (SDS\u2010PAGE) confirmed its size to be 39\u00a0kDa. Kinetic studies revealed that the enzyme has a specific activity of 107.14 \u00b5mol\/min\/mg, a <jats:italic>V<\/jats:italic><jats:sub>max<\/jats:sub> of 25.1\u00a0\u00b5mol\/min, and a <jats:italic>K<\/jats:italic><jats:sub>M<\/jats:sub> of 1.58\u00a0mM. This is the first report on the successful expression of soluble and active recombinant TbPLA<jats:sub>2<\/jats:sub>, which will facilitate the discovery of its specific inhibitors for the development of therapeutics for trypanosomiasis.<\/jats:p>","DOI":"10.1002\/elsc.70005","type":"journal-article","created":{"date-parts":[[2025,3,23]],"date-time":"2025-03-23T04:13:29Z","timestamp":1742703209000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Overproduction and Characterization of Recombinant Soluble <i>Trypanosoma brucei<\/i> Phospholipase A<sub>2<\/sub>"],"prefix":"10.1002","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0696-2713","authenticated-orcid":false,"given":"Oluwafemi Abiodun","family":"Adepoju","sequence":"first","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Nigeria"},{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences University of California San Diego  La Jolla California USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel","family":"Quinnell","sequence":"additional","affiliation":[{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences University of California San Diego  La Jolla California USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Harshverdhan","family":"Sirohi","sequence":"additional","affiliation":[{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences University of California San Diego  La Jolla California USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emmanuel","family":"Amlabu","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Prince Abubakar Audu University  Anyigba Nigeria"},{"name":"Africa Centre of Excellence for Neglected Tropical Diseases and Forensic Biotechnology Ahmadu Bello University  Zaria Nigeria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abdullahi Balarabe","family":"Sallau","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Nigeria"},{"name":"Africa Centre of Excellence for Neglected Tropical Diseases and Forensic Biotechnology Ahmadu Bello University  Zaria Nigeria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abdulrazak","family":"Ibrahim","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Nigeria"},{"name":"Forum for Agricultural Research in Africa (FARA), PMB CT 173, Cantonments  Accra Ghana"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sunday Ene\u2010Ojo","family":"Atawodi","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Federal University Lokoja  Lokoja Kogi Nigeria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohammed Nasiru","family":"Shuaibu","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Nigeria"},{"name":"Africa Centre of Excellence for Neglected Tropical Diseases and Forensic Biotechnology Ahmadu Bello University  Zaria Nigeria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Geoffrey","family":"Chang","sequence":"additional","affiliation":[{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences University of California San Diego  La Jolla California USA"},{"name":"Department of Pharmacology, School of Medicine University of California San Diego  La Jolla California USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emmanuel Oluwadare","family":"Balogun","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Ahmadu Bello University  Zaria Nigeria"},{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences University of California San Diego  La Jolla California USA"},{"name":"Africa Centre of Excellence for Neglected Tropical Diseases and Forensic Biotechnology Ahmadu Bello University  Zaria Nigeria"},{"name":"Department of Biomedical Chemistry, Graduate School of Medicine The University of Tokyo  Hongo Tokyo Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,3,20]]},"reference":[{"unstructured":"WHO Trypanosomiasis Human African (Sleeping Sickness). 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