Pereira Jose H, Goh Ee-Been, Keasling Jay D, Beller Harry R, Adams Paul D
Joint BioEnergy Institute, Emeryville, CA 94608, USA.
Acta Crystallogr D Biol Crystallogr. 2012 Oct;68(Pt 10):1320-8. doi: 10.1107/S0907444912028351. Epub 2012 Sep 18.
Micrococcus luteus is a Gram-positive bacterium that produces iso- and anteiso-branched alkenes by the head-to-head condensation of fatty-acid thioesters [coenzyme A (CoA) or acyl carrier protein (ACP)]; this activity is of interest for the production of advanced biofuels. In an effort to better understand the control of the formation of branched fatty acids in M. luteus, the structure of FabH (MlFabH) was determined. FabH, or β-ketoacyl-ACP synthase III, catalyzes the initial step of fatty-acid biosynthesis: the condensation of malonyl-ACP with an acyl-CoA. Analysis of the MlFabH structure provides insights into its substrate selectivity with regard to length and branching of the acyl-CoA. The most structurally divergent region of FabH is the L9 loop region located at the dimer interface, which is involved in the formation of the acyl-binding channel and thus limits the substrate-channel size. The residue Phe336, which is positioned near the catalytic triad, appears to play a major role in branched-substrate selectivity. In addition to structural studies of MlFabH, transcriptional studies of M. luteus were also performed, focusing on the increase in the ratio of anteiso:iso-branched alkenes that was observed during the transition from early to late stationary phase. Gene-expression microarray analysis identified two genes involved in leucine and isoleucine metabolism that may explain this transition.
藤黄微球菌是一种革兰氏阳性细菌,它通过脂肪酸硫酯(辅酶A(CoA)或酰基载体蛋白(ACP))的头对头缩合产生异支链和反异支链烯烃;这种活性对于先进生物燃料的生产具有重要意义。为了更好地理解藤黄微球菌中支链脂肪酸形成的调控机制,我们测定了FabH(MlFabH)的结构。FabH,即β-酮酰-ACP合酶III,催化脂肪酸生物合成的起始步骤:丙二酰-ACP与酰基-CoA的缩合反应。对MlFabH结构的分析为其对酰基-CoA长度和支链的底物选择性提供了深入见解。FabH结构差异最大的区域是位于二聚体界面的L9环区域,该区域参与酰基结合通道的形成,从而限制了底物通道的大小。位于催化三联体附近的苯丙氨酸336残基似乎在支链底物选择性中起主要作用。除了对MlFabH的结构研究外,我们还进行了藤黄微球菌的转录研究,重点关注在从早期到晚期稳定期的转变过程中观察到的反异支链:异支链烯烃比例的增加。基因表达微阵列分析鉴定出两个参与亮氨酸和异亮氨酸代谢的基因,这可能解释了这种转变。