Chiral separation of β-blockers by high-performance capillary electrophoresis based on non-immobilized cellulase as enantioselective protein

This review gives a survey of different chiral separation principles and their use in high-performance liquid chromatography (HPLC), gas chromatography (GC), supercritical fluid chromatography (SFC), thin-layer chromatography (TLC), capillary electrophoresis (CE) and capillary electrochromatography (CEC) highlighting new developments and innovative techniques. The mechanisms of the different separation principles are briefly discussed and some selected applications are shown.

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“…Fungal cellulase [418] and cellobiohydrolase I (CBH I) [64,[419][420][421] showed remarkable chiral recognition ability for b-blockers.…”

Section: Neutral CD Derivatives a Great Variety Of Neutral Derivativmentioning

confidence: 99%

Chiral separation by chromatographic and electromigration techniques. A Review

Gübitz

Schmid

2001

Biopharm & Drug Disp

225

135

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“…It can be coupled to a solid support (silica or continuous bed) [l(L12] to form a chiral stationary phase (CSP) or used in a carrier-free solution in capillary electrophoresis [13]. A rather broad spectrum of racemic solutes can be resolved, but CBH 1 functions best with amino alcohols such as the drugs used to block [3-adrenergic receptors, so called [3-blockers. In an attempt to localize the chiral site of CBH 1, we found that both the binding and catalytic domains contained enantioselective sites for propranolol, a [3-blocker, but that the major enantioselectivity for propranolol and other solutes could be ascribed to the catalytic domain [14].…”

Section: Introductionmentioning

confidence: 99%

The active sites of cellulases are involved in chiral recognition: a comparison of cellobiohydrolase 1 and endoglucanase 1

et al. 1996

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The cellulases ceHobiohydrolase 1 (CBH 1) and endoglucanase 1 (EG 1) from the fungus Trichoderma reesei are closely related with 40% sequence identity and very similar in structure. In CBH 1 the active site is enclosed by long loops and some antiparallel ~strands forming a 40 A long tunnel, whereas in EG 1 part of those loops are missing so that the enzyme has a more common active site groove. Both enzymes were immobilized on silica and these materials were used as chiral stationary phases for chromatographic separation of the enantiomers of two chiral drugs, propranolol and alprenolol. The CBH 1 phase showed much better resolution than did the EG 1 phase, suggesting that the tunnel structure of the protein may play an important role in the chiral separation. The chiral compounds were found to be competitive inhibitors of both enzymes when p-nitrophenyl lactoside (pNPL) was used as substrate. (S)-enantiomers showed stronger inhibitory effects and also longer retention time on the stationary phases than the (R)-enantiomers. The consistency between kinetic data and retention on the stationary phases clearly shows that the enzymatically active sites of CBH 1 and EG 1 are involved in chiral recognition.

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“…The PFT, first introduced by Valtcheva et al 182 and further modified by Tanaka et al, 183 involves the filling of a discrete portion of the CE capillary with BGE containing a chiral selector (i.e., partial filling). This method is particularly adapted in chiral CE2MS and presents major advantages over the traditional approach.…”

Section: (B) Partial-filling Techniquementioning

confidence: 99%