Determination of optimal range of post-induction temperature for production of soluble recombinant bromelain in Escherichia coli using one-factor-at-a-time (OFAT) approach
In this work, a one-factor-at-a-time (OFAT) experiment for determining the optimal preliminary range of post-induction temperature (OC), before subsequent optimization of recombinant bromelain production using Response Surface Methodology (RSM), are discussed. Bromelain from Ananas comosus, in na...
| Main Authors: | , |
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| Format: | Book Chapter |
| Language: | English |
| Published: |
IIUM Press
2011
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/20920/ http://irep.iium.edu.my/20920/ http://irep.iium.edu.my/20920/1/determination_of_optimal_-_C5.pdf |
| Summary: | In this work, a one-factor-at-a-time (OFAT) experiment for determining the optimal
preliminary range of post-induction temperature (OC), before subsequent optimization of
recombinant bromelain production using Response Surface Methodology (RSM), are
discussed. Bromelain from Ananas comosus, in nature, being a eukaryotic plant-origin
proteolytic enzyme, has been subjected to post-translational modification such as
glycosylation and presence of native molecular chaperones for aiding the protein folding.
However, attempts to express this enzyme in Escherichia coli might incur undesirable
potential drawback, which is, high tendency of the recombinant bromelain to express in
the form of inactive insoluble wasted fractions known as inclusion bodies. Induction
temperature has been known to be one of the controllable factors which could minimize
the formation of inclusion bodies. Here, post-induction temperatures between 21 - 41°C
were studied and their effects on the recombinant bromelain specific activity (U/mg)
were evaluated. We found that post-induction temperature around 26°C marked highest
value of specific activity (~14 U/mg) and therefore, shall be used as a center-point for the
next RSM analysis. |
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