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- Title
Expression of soluble moloney murine leukemia virus-reverse transcriptase in Escherichia coli BL21 star (DE3) using autoinduction system.
- Authors
Handayani, Christina Vivid; Laksmi, Fina Amreta; Andriani, Ade; Nuryana, Isa; Mubarik, Nisa Rachmania; Agustriana, Eva; Dewi, Kartika Sari; Purnawan, Awan
- Abstract
Autoinduction systems in Escherichia coli can control the production of proteins without the addition of a particular inducer. In the present study, we optimized the heterologous expression of Moloney Murine Leukemia Virus derived Reverse Transcriptase (MMLV-RT) in E. coli. Among 4 autoinduction media, media Imperial College resulted the highest MMLV-RT overexpression in E. coli BL21 Star (DE3) with incubation time 96 h. The enzyme was produced most optimum in soluble fraction of lysate cells. The MMLV-RT was then purified using the Immobilized Metal Affinity Chromatography method and had specific activity of 629.4 U/mg. The system resulted lower specific activity and longer incubation of the enzyme than a classical Isopropyl ß-D-1-thiogalactopyranoside (IPTG)-induction system. However, the autoinduction resulted higher yield of the enzyme than the conventional induction (27.8%). Techno Economic Analysis revealed that this method could produce MMLV-RT using autoinduction at half the cost of MMLV-RT production by IPTG-induction. Bioprocessing techniques are necessary to conduct to obtain higher quality of MMLV-RT under autoinduction system.
- Subjects
MOUSE leukemia; MOUSE leukemia viruses; ESCHERICHIA coli; REVERSE transcriptase; AFFINITY chromatography; INDUSTRIAL costs
- Publication
Molecular Biology Reports, 2024, Vol 51, Issue 1, p1
- ISSN
0301-4851
- Publication type
Article
- DOI
10.1007/s11033-024-09583-6