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Title
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Expression of soluble and catalytically active plant (monocot) beta-glucosidases in E. coli.
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Authors
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M.Cicek,
A.Esen.
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Ref.
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Biotechnol Bioeng, 1999,
63,
392-400.
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PubMed id
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Abstract
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Complementary DNAs encoding mature beta-glucosidase proteins Glu1 and Glu2 of
maize were amplified by the polymerase chain reaction (PCR) and cloned into the
expression vector pET21a. Both Glu1 and Glu2 isozymes were expressed in high
yield ( approximately 3.8% of the total soluble protein and 32% of the total
expressed protein) in E. coli. Recombinant enzymes were active on a variety of
artificial and natural substrates at levels similar to those of their native
counterparts isolated from maize seedlings. Western blot analysis confirmed that
both recombinant isozymes were immunoreactive with maize anti-beta-glucosidase
sera and their molecular sizes were identical to those of the native maize Glu1
and Glu2 isozymes. Zymogram assays in native gels revealed that recombinant
enzymes had the same electrophoretic mobility and substrate specificity as their
native counterparts.
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