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Expression and functional characterization of a C-7 cholesterol desaturase from Tetrahymena thermophila in an insect cell line.

Steroids. 2015-04; 
PoklepovichTomas J,UrtasunNicolás,MirandaMaría V,NusblatAlejandro D,NudelCla
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Custom Vector Construction … template (Bentzon-Tilia et al, 2014) The qPCR standards were synthesized and cloned into the pUC57 vector at GenScript (Piscataway, NJ, USA) and linearized by HindIII digestion Reaction mixtures, thermal cycling settings … Get A Quote

摘要

Tetrahymena thermophila transforms exogenous cholesterol into pro-vitamin D3 (7-dehydrocholesterol) with remarkable efficiency in a one-step reaction carried out by a C-7 cholesterol desaturase. The enzyme DES7 is encoded by the gene TTHERM_00310640, identified with RNAi and gene knock-out experiments, but has not yet been heterologously expressed actively in any organism. A model derived from its amino acid sequence classified DES7p as a Rieske-type oxygenase with transmembrane localization. The protein has catalytic activity, sequence and topological similarity to DAF-36/Neverland proteins involved in the synthesis of steroid hormones in insects and nematodes. Due to their structural and functional simi... More

关键词

C-7 cholesterol desaturase,Cholesterol,Oxygenase,Pro-vitamin D ,Sf9 cell line,Tetrahymena thermop