Part:BBa_K2983081
FadX of Punica granatum expression cassette under the control of pTef1 (BBa_K2983052) promoter
FadX of Punica granatum expression cassette under the control of pTef1 (BBa_K2983052) promoter
Usage and Biology
Bioproduction
Conjugated linolenic acids (CLnAs) are synthetized by bifunctional fatty acid conjugase / desaturase (FadX) enzymes from linoleic acid (incorporated into phosphatidylcholine). The sequences of the enzymes catalyzing the synthesis of 3 of the 7 known CLnAs have been described in the literature and their activities are specific to the position and the stereochemistry of the double bonds:
- punicic acid, C18:3 (9Z, 11E, 13Z) is synthesized by EC: 1.14.19.16 [1,2].
- α-calendic acid, C18:3 (8E, 10E, 12Z), is synthesized by EC: 1.14.19.14 [3,4].
- α-oleosteaic acid, C18:3 (9Z, 11E, 13E) is synthesized by EC: 1.14.19.33 [5].
Design
Linoleic acid, C18:2 (9Z,12Z), the substrate of FadX enzymes, is a natural metabolite for our chassis Yarrowia lipolytica. Thus, to convert it into punicic acid, only the presence of a EC: 1.14.19.16 enzyme is necessary (Figure 1). Two EC: 1.14.19.16 enzymes were described in the literature: one from pomegranate / Punica granatum (Pg-FadX, BBa_K2983061) and another one from the chinese cucumber / chinese snake gourd / Trichosanthes kirilowii (Tk-FadX, BBa_K2983062).
To achieve a sustainable bioproduction of such fatty acids in order to limit environmental and economical problems, we decided to use as a biological chassis the oleaginous yeast Yarrowia lipolytica. This species has already proven its effectiveness for the production of fatty acids, thanks to its highly developed lipid metabolism [6-8]. As a proof of concept of our project, we decided to focus on one of those CLnAs, the punicic acid, that has interesting properties such as anti-obesity, anti-inflammatory, anti-cancer, anti-diabetes activities [9].
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 192
Illegal PstI site found at 157 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 192
Illegal PstI site found at 157 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 192
Illegal PstI site found at 157 - 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 192
Illegal PstI site found at 157 - 1000COMPATIBLE WITH RFC[1000]
| None |