Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method

This study describes the development of an intensified process that enables high level production of recombinant core streptavidin (core SAV), a non-glycosylated tetrameric protein utilised in a wide range of applications. Due to widespread utility and commercial importance of SAV, recombinant SA...

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Main Author: Chua, Leong Huat
Format: Thesis
Language:English
Published: 2019
Subjects:
Online Access:http://eprints.usm.my/61953/1/intensification%20of%20core%20streptavidin.pdf
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author Chua, Leong Huat
author_facet Chua, Leong Huat
author_sort Chua, Leong Huat
collection USM
description This study describes the development of an intensified process that enables high level production of recombinant core streptavidin (core SAV), a non-glycosylated tetrameric protein utilised in a wide range of applications. Due to widespread utility and commercial importance of SAV, recombinant SAV production has been studied extensively. However, in most studies, recombinant SAV was expressed in a soluble form, resulting in in vivo sequestration of biotin. Biotin sequestration is detrimental to cell growth and results in low volumetric yield. To improve volumetric yield, in this study, core SAV was expressed as inclusion bodies (IBs) followed by induction at highcell- density. A pH-stat fed-batch feeding strategy was employed to cultivate host cells to high-cell-density, the effect of induction at different cell densities (OD 20, 60 and 100) on volumetric and specific yield were then studied. Highest volumetric yield of core SAV (1.55 g L"1) was obtained from induction at OD 100.
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spelling usm.eprints-619532025-03-03T01:47:42Z http://eprints.usm.my/61953/ Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method Chua, Leong Huat QR75-99.5 Bacteria This study describes the development of an intensified process that enables high level production of recombinant core streptavidin (core SAV), a non-glycosylated tetrameric protein utilised in a wide range of applications. Due to widespread utility and commercial importance of SAV, recombinant SAV production has been studied extensively. However, in most studies, recombinant SAV was expressed in a soluble form, resulting in in vivo sequestration of biotin. Biotin sequestration is detrimental to cell growth and results in low volumetric yield. To improve volumetric yield, in this study, core SAV was expressed as inclusion bodies (IBs) followed by induction at highcell- density. A pH-stat fed-batch feeding strategy was employed to cultivate host cells to high-cell-density, the effect of induction at different cell densities (OD 20, 60 and 100) on volumetric and specific yield were then studied. Highest volumetric yield of core SAV (1.55 g L"1) was obtained from induction at OD 100. 2019-05 Thesis NonPeerReviewed application/pdf en http://eprints.usm.my/61953/1/intensification%20of%20core%20streptavidin.pdf Chua, Leong Huat (2019) Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method. Masters thesis, Universiti Sains Malaysia.
spellingShingle QR75-99.5 Bacteria
Chua, Leong Huat
Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method
title Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method
title_full Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method
title_fullStr Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method
title_full_unstemmed Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method
title_short Intensification Of Core Streptavidin Production Through High-Cell-Density Cultivation Of Recombinant Escherichia Coli And A Temperature-Based Refolding Method
title_sort intensification of core streptavidin production through high cell density cultivation of recombinant escherichia coli and a temperature based refolding method
topic QR75-99.5 Bacteria
url http://eprints.usm.my/61953/1/intensification%20of%20core%20streptavidin.pdf
work_keys_str_mv AT chualeonghuat intensificationofcorestreptavidinproductionthroughhighcelldensitycultivationofrecombinantescherichiacoliandatemperaturebasedrefoldingmethod