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.
Abstract
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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