کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1178804 962726 2011 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Expression, purification and characterization of two Clostridium acetobutylicum flavodoxins: Potential electron transfer partners for CYP152A2
کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Expression, purification and characterization of two Clostridium acetobutylicum flavodoxins: Potential electron transfer partners for CYP152A2
چکیده انگلیسی

Two flavodoxin genes from Clostridium acetobutylicum, CacFld1 (CAC0587) and CacFld2 (CAC3417), were expressed in Escherichia coli and investigated for their ability to support activity of CYP152A2, a fatty acid hydroxylase from C. acetobutylicum. E. coli flavodoxin reductase (FdR) was used as a redox partner, since flavodoxin reductase CacFdR (CAC0196) from C. acetobutylicum could not be purified in a functional form. CacFld1 was shown to accept electrons from FdR and transfer them to CYP152A2. Since H2O2 was generated by uncoupling at different stages of the reconstituted electron transfer chain, catalase was used as H2O2 scavenger in order to exclude peroxygenation by CYP152A2. The reconstituted P450 system with CacFld1 and FdR oxidized myristic acid with a KM of 137 μM and a kcat of 36 min−1. Furthermore, the hydroxylase activity of CYP152A2 towards myristic acid with CacFld1 was 17-fold higher than without CacFld1. Along with CYP152A2 and a physiological flavodoxin reductase, CacFld1 is therefore likely to be involved in oxygen detoxification in C. acetobutylicum. Flavodoxin CacFld2 did not accept electrons from NADPH-reduced FdR, though it cannot be excluded as a candidate redox partner for CYP152A2 in the presence of an appropriate physiological reductase.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics - Volume 1814, Issue 1, January 2011, Pages 257–264
نویسندگان
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