M.I.E.R. (A new instrument for proteomics)

Pier Carlo Montecucchi pcmontecucchi at compuserve.com
Wed Nov 15 05:39:35 EST 2000


"M.I.E.R." (=Multicompartment Immobilized Enzyme Reactor) is an
isoelectrically enzyme reactor that integrates catalytic conversion, product
separation, product concentration and catalytic recovery into a single
operation.

THE INSTRUMENT IS DESIGNED FOR :

(i)    the purification of biomolecules obtainable from a catalytic
bioconversion process;
(ii)   designing and testing enzymatic, kinetic assays for high-throughput
screening;
(iii)  protein purification;
(iv)  protein primary structure determination;
(v)   understanding specific metabolic functions of protein isoforms.

REALIZED APPLICATIONS:

For enzymatic catalysis:

(i)    Bioconversion of charged  penicillin G by penicillin G acylase

(ii)   Histidine decarboxylase reactor

(iii)   Production of pure D-phenylglycine by penicillin G Acylase

(iv)   Thrombin reactor

(v)    Thrombin digestion of Gst-fusion protein (Gst-Tat) and subsequent
purification of cleaved products. Tat, released upon thrombin hydrolysis, is
harvested in a pure form in the pH region 9 to 11; Gst in the pH region  <5

(vi)   Trypsin reactor : The tryptic digestion of a milk beta-casein
releases the peptides with different  biological activities either directly
[peptide beta(1-25) and beta (33-48) with anticariogenic activity and
peptide beta (114 - 169)] or indirectly as a precursor [peptide beta (49-97)
containing in its sequence peptide beta (60-66) with casomorphine activity
and peptide beta (63-68) with antihypertensive activity]

(vii)      Urease reactor

For protein purification:

 (i)     Purification of chicken liver basic FABS (fatty acid binding
protein)

(ii)    Analysis of cross-linked human hemoglobin by conventional
isoelectric focusing, immobilized pH gradients, capillary electrophoresis,
and mass spectrometry.


For more details and the scientific references on the topic, contact:
Montegen (phone: 212-208-2647; fax: 212-208-2648; Email: info at montegen.com;
URL: www.montegen.com)









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