کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
26478 43954 2015 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Expression systems for soluble metal-dependent formate dehydrogenase
ترجمه فارسی عنوان
سیستم های بیان برای فرمیت دی هیدروژناز متابولیزه فلزی
کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
چکیده انگلیسی


• We successfully established novel expression systems, which enables to produce soluble metal-dependent formate dehydrogenases more easily.
• We successfully converted a membrane bound metal-dependent formate dehydrogenase from Escherichia coli into a soluble enzyme, and demonstrated that the newly created formate dehydrogenase can be purified under aerobic conditions.
• We succeeded in the improvement of metal-dependent formate dehydrogenase by molecular evolution procedure.
• We successfully create Desulfovibrio mutants expressing a soluble metal-dependent formate dehydrogenases tagged with Strep-tagII.

Molybdenum or tungsten-dependent formate dehydrogenases (FDH) can reduce carbon dioxide (CO2) to formate under ordinary conditions, and therefore, are considered promising catalysts for CO2 fixation. However, to our knowledge, no study on the modification of metal-dependent FDHs has been published, likely because of a lack of convenient expression systems for the recombinant enzymes. We attempted to establish an methodology for the preparation and modification of soluble oxygen-tolerant metal-dependent FDHs, for the following three strategies: (1) Escherichia coli FDH is converted from a membrane bound protein into a soluble protein by deleting the C-terminal membrane-anchor of small subunit (FdoH) and the whole membrane subunit (FdoI) and expressed homologously in E. coli; (2) originally soluble FDHs from Desulfovibrio are expressed heterologously in E. coli; and (3) Desulfovibrio FDHs are genetically engineered by the homologous gene recombination method and expressed homologously in Desulfovibrio. We successfully established the expression systems for (1) and (3), and succeeded in purification of the soluble FDHs with a tag by using affinity columns.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Photochemistry and Photobiology A: Chemistry - Volume 313, 1 December 2015, Pages 154–162
نویسندگان
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