Abstract
A new and highly sensitive method was developed for the identification
of hydantoinases on acrylamide gels. For this purpose, cell-lysates from
different natural isolates are subjected on PAGE under non-denaturating
conditions. The respective localisation of the hydantoinase is obtained
by in situ product precipitation during the reverse enzyme reaction: in
contrast to the used substrate (N-carbamoyltryptophan), the product
(indolylmethylhydantoin) is barely soluble and gives a dense
precipitation dot caused by crystallisation of the product inside of the
polyacrylamide gel at the position corresponding to the location of the
enzyme. This method can also be used for the subsequent differentiation
between L- and D-selective hydantoinases, since L- or
D-carbamoyltryptophan is used as substrate.
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