M.I.E.R. / Enzymatic catalysis - Protein purification

Pier carlo Montecucchi pcmontecucchi at compuserve.com
Mon Oct 22 08:43:07 EST 2001


The Multicompartment Immobilized Enzyme Reactor (M.I.E.R.) is an
isoelectrically trapped enzyme reactor that integrates catalytic conversion,
product separation, product concentration and catalyst recovery into a
single operation. The instrument is modular and it is built to comprise 8
flow-through (recycling) chambers: one anodic, one cathodic, one for the
enzyme reactor and five chambers for collecting any desired product of the
enzyme reaction. The enzyme reactor operates under an electric field, which
is used to keep the enzyme confined into the reaction chamber (via an
isoelectric mechanism) and to drive in and out of the reaction chamber any
charged substrate, product or co-factor.
The enzyme is immobilized between two isoelectric membranes encompassing the
isoelectric point of the enzyme and the pH of its optimum activity. Since it
is not covalently bound to any support but it is free in solution, its
kinetic parameters are identical with or very close to the parameters of the
enzyme in solution. Each compartment has a membrane pump for circulating the
reagent flow through the cells and the reservoirs. Sampling the reaction is
always possible during the process. The inside temperature of the instrument
is regulated by a Peltier element from 8 to 48 C. Handling of the cell
tubing connections and washing of the machine are user-friendly operations.

The following are examples of the realized reactors:
· Bioconversion of charged penicillin G by penicillin G acylase
· Production of pure D-phenylglycine by penicillin G acylase
· Urease reactor
· Trypsin reactor (for peptide generation)
· Thrombin reactor (digestion of Gst-fusion protein and purification of
cleaved product
· Histidine decarboxylase reactor (production of histamine)

On specific requests, we can offer "ad hoc" services to clients interested
in the purification of biomolecules obtainable from a catalytic
bioconversion process.

For more information, contact: MONTEGEN, 540 Beverly Ct. - Suite 1 - P.O.
Box 7357 - Tallahassee, FL 32314 - 7357 U.S.A.; Email: info at montegen.com;
URL: www.montegen.com






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