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Mol Biol Rep ; 51(1): 628, 2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38717629

RESUMO

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.


Assuntos
Escherichia coli , Vírus da Leucemia Murina de Moloney , DNA Polimerase Dirigida por RNA , Escherichia coli/genética , Escherichia coli/metabolismo , Vírus da Leucemia Murina de Moloney/genética , Vírus da Leucemia Murina de Moloney/enzimologia , DNA Polimerase Dirigida por RNA/metabolismo , DNA Polimerase Dirigida por RNA/genética , Isopropiltiogalactosídeo/farmacologia , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/genética , Meios de Cultura
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