摘要 : Linoleate 13-lipoxygenase from Burkholderia thailandensis was expressed in Escherichia coli for the production of 13-hydroxyoctadecadienoic acid (13-HODE), an antiseptic emulsifier. Linoleate 13-lipoxygenase in cells had higher th... 展开 Linoleate 13-lipoxygenase from Burkholderia thailandensis was expressed in Escherichia coli for the production of 13-hydroxyoctadecadienoic acid (13-HODE), an antiseptic emulsifier. Linoleate 13-lipoxygenase in cells had higher thermal stability than the purified enzyme. To increase 13-HODE production, recombinant cells were permeabilized by solvents, detergents, salts, and other chemicals. The enzymatic activity in cells was the highest for permeabilized cells treated with 0.5 M NaCl among the permeabilizers tested. The optimal reaction conditions for the production of 13-HODE from linoleic acid by permeabilized cells treated with 0.5 M NaCl were at pH 7.5, 25 A degrees C, 20 g/l linoleic acid, 15 g/l cells, 0.15 mM Cu2+, and 6 % (v/v) methanol in a 100-ml baffled flask containing a 5-ml working volume with agitation at 200 rpm. Under these conditions, permeabilized cells produced 15.8 g/l 13-HODE after 30 min with a conversion yield of 79 % (w/w) and a productivity of 31.6 g/l/h. The conversion yield and productivity of permeabilized cells for 13-HODE production were higher than those of purified and crude enzymes as well as nonpermeabilized cells. Therefore, permeabilized cells were efficient biocatalysts for 13-HODE production. To the best of our knowledge, this is the first report of the production of 13-HODE using cells. 收起