Is there any way to cut down the excess (not required) amino acids in a single chain polypeptide enzyme without disturbing its active site ?
It is the question what is the excess which is not required! Can you define it? The spatial configuration of the active site is dependent on peptide regions not close to it which respect to the amino acid chain. 'Remote' regions may determine the right configuration influencing substrate access and proper substrate orientation. If you are sure that certain C-terminal or N-terminal sequences are unnecessary to maintain the enzymatic activities without altering the enzyme characteristis (KM, vmax) you will possibly find a specific protease to shorten the peptide.
Peter
Hi there,
If you have a straight forward assay in hand for screening your enzyme activity, you may use an iterative truncation library approach to generate the whole set of truncation variants.
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