Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli
The chicken-type lysozyme of the insect Spodoptera litura (SLLyz) is a polypeptide of 121 amino acids containing four disulfide bridges and 17 rare codons and participates in innate defense as an anti-bacterial enzyme. The recombinant S. litura lysozyme (rSLLyz) expressed as a C-terminal fusion prot...
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Pontificia Universidad Católica de Valparaíso
2011
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oai:scielo:S0717-345820110003000062011-09-23Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coliKim,Jong-WanYoe,JeehyunLee,Gil HoYoe,Sung Moon antibacterial activity inclusion body lysozyme on-column refolding recombinant expression Spodoptera litura The chicken-type lysozyme of the insect Spodoptera litura (SLLyz) is a polypeptide of 121 amino acids containing four disulfide bridges and 17 rare codons and participates in innate defense as an anti-bacterial enzyme. The recombinant S. litura lysozyme (rSLLyz) expressed as a C-terminal fusion protein with glutathione S-transferase (GST) in Rosetta(DE3) Singles. The protein was produced as an inclusion body which was solubilized in 8 M urea, renatured by on-column refolding, and purified by reversed-phase chromatography to 95% purity. The purified rSLLyz demonstrated antibacterial activity against B. megaterium confirmed by inhibition zone assay. The overexpression and refolding strategy described in this study will provide a reliable technique for maximizing production and purification of proteins expressed as inclusion bodies in E. coli.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.14 n.3 20112011-05-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300006en |
| institution |
Scielo Chile |
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Scielo Chile |
| language |
English |
| topic |
antibacterial activity inclusion body lysozyme on-column refolding recombinant expression Spodoptera litura |
| spellingShingle |
antibacterial activity inclusion body lysozyme on-column refolding recombinant expression Spodoptera litura Kim,Jong-Wan Yoe,Jeehyun Lee,Gil Ho Yoe,Sung Moon Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli |
| description |
The chicken-type lysozyme of the insect Spodoptera litura (SLLyz) is a polypeptide of 121 amino acids containing four disulfide bridges and 17 rare codons and participates in innate defense as an anti-bacterial enzyme. The recombinant S. litura lysozyme (rSLLyz) expressed as a C-terminal fusion protein with glutathione S-transferase (GST) in Rosetta(DE3) Singles. The protein was produced as an inclusion body which was solubilized in 8 M urea, renatured by on-column refolding, and purified by reversed-phase chromatography to 95% purity. The purified rSLLyz demonstrated antibacterial activity against B. megaterium confirmed by inhibition zone assay. The overexpression and refolding strategy described in this study will provide a reliable technique for maximizing production and purification of proteins expressed as inclusion bodies in E. coli. |
| author |
Kim,Jong-Wan Yoe,Jeehyun Lee,Gil Ho Yoe,Sung Moon |
| author_facet |
Kim,Jong-Wan Yoe,Jeehyun Lee,Gil Ho Yoe,Sung Moon |
| author_sort |
Kim,Jong-Wan |
| title |
Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli |
| title_short |
Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli |
| title_full |
Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli |
| title_fullStr |
Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli |
| title_full_unstemmed |
Recombinant expression and refolding of the c-type lysozyme from Spodoptera litura in E. coli |
| title_sort |
recombinant expression and refolding of the c-type lysozyme from spodoptera litura in e. coli |
| publisher |
Pontificia Universidad Católica de Valparaíso |
| publishDate |
2011 |
| url |
http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000300006 |
| work_keys_str_mv |
AT kimjongwan recombinantexpressionandrefoldingofthectypelysozymefromspodopteraliturainecoli AT yoejeehyun recombinantexpressionandrefoldingofthectypelysozymefromspodopteraliturainecoli AT leegilho recombinantexpressionandrefoldingofthectypelysozymefromspodopteraliturainecoli AT yoesungmoon recombinantexpressionandrefoldingofthectypelysozymefromspodopteraliturainecoli |
| _version_ |
1718441837412220928 |