Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628, BC, Delft, the Netherlands.
Appl Microbiol Biotechnol. 2010 Jun;87(2):679-90. doi: 10.1007/s00253-010-2626-z. Epub 2010 May 7.
A transcriptomics and proteomics approach was employed to study the expression changes associated with p-hydroxybenzoate production by the engineered Pseudomonas putida strain S12palB1. To establish p-hydroxybenzoate production, phenylalanine-tyrosine ammonia lyase (pal/tal) was introduced to connect the tyrosine biosynthetic and p-coumarate degradation pathways. In agreement with the efficient p-hydroxybenzoate production, the tyrosine biosynthetic and p-coumarate catabolic pathways were upregulated. Also many transporters were differentially expressed, one of which--a previously uncharacterized multidrug efflux transporter with locus tags PP1271-PP1273--was found to be associated with p-hydroxybenzoate export. In addition to tyrosine biosynthesis, also tyrosine degradative pathways were upregulated. Eliminating the most prominent of these resulted in a 22% p-hydroxybenzoate yield improvement. Remarkably, the upregulation of genes contributing to p-hydroxybenzoate formation was much higher in glucose than in glycerol-cultured cells.
采用转录组学和蛋白质组学方法研究了与工程假单胞菌 S12palB1 生产对羟基苯甲酸相关的表达变化。为了建立对羟基苯甲酸的生产,引入了苯丙氨酸-酪氨酸氨裂解酶(pal/tal)将酪氨酸生物合成和对香豆酸降解途径连接起来。与高效生产对羟基苯甲酸一致,酪氨酸生物合成和对香豆酸分解代谢途径被上调。许多转运蛋白也表现出差异表达,其中一个——以前未被表征的多药外排转运蛋白,带有基因标记 PP1271-PP1273——与对羟基苯甲酸的外排有关。除了酪氨酸生物合成,酪氨酸降解途径也被上调。消除其中最显著的途径可使对羟基苯甲酸的产量提高 22%。值得注意的是,在葡萄糖培养的细胞中,参与对羟基苯甲酸形成的基因的上调水平明显高于甘油培养的细胞。