Genetic engineering techniques were employed for development of efficient expression systems of recombinant human acetylcholinesterase (rHuAChE). Human cell lines allowing production of 10-20 mg/liter of fully-active enzyme were established. Biotechnological processes for large scale production of rHuAChE from these cells were evaluated in multitray systems, fixed-bed reactors and microcarrier cultures. The latter proved to be the most promising, exhibiting system productivity of 6-10 mg/liter/day. rHuAChE is secreted from the recombinant ...
Combination of recombinant DNA technology, kinetic studies and molecular modeling, are being employed in the design of novel biocatalysts with potential pharmacological use against organophosphate (OP) poisoning. Initial efforts were directed towards unravelling the intricate architecture of the catalytic machinery of human acetylcholinesterase (HuAChE). Thus, analysis of over 40 site directed mutants of HuAChE identified: (a) The esteratic site - Ser2O3, His447 and Glu334 constitute the catalytic triad. Glu2O2 at ...
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