Reconstitution of Escherichia coli Fatty Acid Biosynthesis Reaction In vitro
- VernacularTitle:大肠杆菌(Escherichia coli)体外脂肪酸合成反应的重建
- Author:
Saixiang FENG
;
Lei ZHU
;
Biao LUO
;
Yirong SUN
;
Haihong WANG
- Publication Type:Journal Article
- Keywords:
fatty acid synthesis of bacteria, acyl-ACP, native polyacrylamide gels
- From:
Progress in Biochemistry and Biophysics
2006;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Seven genes (fabD, fabG, fabH, fabA, fabZ, fabB and fabI) of E.coli fatty acid biosynthetic enzymes were cloned by PCR amplifying and appropriate expression vectors were constructed. Under induction assay expression of plasmid encoded proteins was carried out in strain BL21(DE3) and seven enzymes were purified using Ni-NTA agarose resin. In the absence of [2-14C] malonyl-CoA fatty acid synthetic reaction was reconstituted in vitro by adding seven enzymes and co-factors. And several model reactions were established for identification of special fatty acid biosynthetic enzymes. Meanwhile Clostridium acetobutylicium FabZ function was characterized by this method.
