Nickel-nitriloacetic acid

The chemical modification of MscL requires a free cysteine residue at a critical part of the channel, namely the 22nd amino acid. A 6-histidine(His) tag at the C-terminal of the protein allows a one-step isolation of MscL, which is >98% pure as analyzed by SDS-PAGE and N-terminal sequencing. The protein can be labeled in its pure isolated form, or alternatively, in its nickel-nitriloacetic acid (Ni-NTA) column-attached form. ESI-MS is a method to follow the labeling conditions and efficiency. After the protein is chemically labeled and reconstituted into desired liposomes, a simple fluorescent, dye efflux experiment is used to test the triggered liposomal release. 

For the in vitro screening of enzyme pairs and enzymatic pathways, immobilized enzyme microreactor systems (lEMR) were established. This includes the reversible immobilization of His -tagged enzymes via nickel-nitriloacetic acid (Ni-NTA) linkage on the surface-derivatized silica. First of all the kinetic parameters for a new enzymatic pathway have to be defined via a transketolase and a w-transaminase in a continuous flow system for a two-step asymmetric synthesis of chiral amino alcohols [147]. The apparenf value is found to be flow rate dependent with different flow rates ranging from 2 to 30 j1/min. The turnover rate was also found to be lower than that in solu-... 

Nickel, sepn. of plus cobalt, (cm) 583 Nickel ware 96 Nioxime 439, 474 NITA (nitriloacetic acid) 57 Nitrates, D. of as nitron nitrate, (g) 484 by ammonium iron(II) sulphate, (am)... 

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