García-Maroto Federico, Mañas-Fernández Aurora, Garrido-Cárdenas José A, Alonso Diego López
Area de Bioquímica, Facultad de Ciencias Experimentales, Universidad de Almería, 04120 Almería, Spain.
Phytochemistry. 2006 Mar;67(6):540-4. doi: 10.1016/j.phytochem.2005.12.005. Epub 2006 Feb 7.
Echium (Boraginaceae) species from the Macaronesian islands exhibit an unusually high level of gamma-linolenic acid (18:3n-6; GLA) and relatively low content of octadecatetraenoic acid (18:4n-3; OTA) in the seed, while the amounts of both fatty acids in their Continental (European) relatives are rather similar. We have tested the hypothesis of whether a different specificity of the acyl-Delta(6)-desaturases (D6DES) towards their respective usual substrates, linoleic acid (18:2n-6; LA) for GLA and alpha-linolenic acid (18:3n-3; ALA) for OTA, was partly responsible for this composition pattern. To this aim we have expressed in yeast the coding sequences of the D6DES genes for the Continental species Echium sabulicola, and the Macaronesian Echium gentianoides. When the yeast cultures are supplemented with the two fatty acid substrates (LA and ALA), a similar utilization of both compounds was found for the D6DES of E. sabulicola, while a preference for LA over ALA was observed for the enzyme of E. gentianoides. This substrate preference must contribute to the increased accumulation of GLA in the seeds of the Macaronesian Echium species. Comparison among the amino acid sequences of these desaturases and other related enzymes, allowed us the discussion about the possible involvement of some specific positions in the determination of substrate specificity.
来自马卡罗尼西亚群岛的蓝蓟属(紫草科)植物种子中γ-亚麻酸(18:3n-6;GLA)含量异常高,而十八碳四烯酸(18:4n-3;OTA)含量相对较低,而它们在欧洲大陆的近缘种中这两种脂肪酸的含量相当相似。我们检验了酰基-Δ(6)-去饱和酶(D6DES)对其各自常见底物(用于合成GLA的亚油酸(18:2n-6;LA)和用于合成OTA的α-亚麻酸(18:3n-3;ALA))的不同特异性是否部分导致了这种组成模式的假设。为此,我们在酵母中表达了大陆种蓝蓟(Echium sabulicola)和马卡罗尼西亚种龙胆叶蓝蓟(Echium gentianoides)的D6DES基因编码序列。当向酵母培养物中添加两种脂肪酸底物(LA和ALA)时,发现蓝蓟(E. sabulicola)的D6DES对这两种化合物的利用情况相似,而龙胆叶蓝蓟(E. gentianoides)的酶对LA的偏好高于ALA。这种底物偏好必定有助于马卡罗尼西亚蓝蓟属植物种子中GLA积累的增加。通过比较这些去饱和酶和其他相关酶的氨基酸序列,我们可以讨论一些特定位置在决定底物特异性方面可能的作用。