Laboratory of Chemical Biotechnology, TU Dortmund University, Emil-Figge-Str. 66, Dortmund, D-44227, Germany.
Environ Microbiol. 2015 Jan;17(1):156-70. doi: 10.1111/1462-2920.12537. Epub 2014 Jul 15.
The natural ability of Pseudomonas taiwanensis VLB120 to use xylose as sole carbon and energy source offers a high potential for sustainable industrial biotechnology. In general, three xylose assimilation routes are reported for bacteria. To elaborate the metabolic capacity of P. taiwanensis VLB120 and to identify potential targets for metabolic engineering, an in silico/in vivo experiment was designed, allowing for discrimination between these pathways. Kinetics of glucose and xylose degradation in P. taiwanensis VLB120 was determined and the underlying stoichiometry was investigated by genome-based metabolic modelling and tracer studies using stable isotope labelling. Additionally, reverse transcription quantitative polymerase chain reaction experiments have been performed to link physiology to the genomic inventory. Based on in silico experiments, a labelling strategy was developed, ensuring a measurable and unique (13) C-labelling distribution in proteinogenic amino acids for every possible distribution between the different xylose metabolization routes. A comparison with in vivo results allows the conclusion that xylose is metabolized by P. taiwanensis VLB120 via the Weimberg pathway. Transcriptomic and physiological studies point to the biotransformation of xylose to xylonate by glucose dehydrogenase. The kinetics of this enzyme is also responsible for the preference of glucose as carbon source by cells growing in the presence of glucose and xylose.
恶臭假单胞菌 VLB120 能够将木糖作为唯一的碳源和能源,这为可持续工业生物技术提供了巨大的潜力。一般来说,细菌中有三种木糖同化途径。为了详细阐述恶臭假单胞菌 VLB120 的代谢能力,并确定代谢工程的潜在目标,设计了一项计算机模拟/体内实验,以便对这些途径进行区分。通过基于基因组的代谢建模和使用稳定同位素标记的示踪研究,确定了恶臭假单胞菌 VLB120 中葡萄糖和木糖降解的动力学,并研究了其基础化学计量学。此外,还进行了逆转录定量聚合酶链反应实验,将生理学与基因组库存联系起来。基于计算机模拟实验,开发了一种标记策略,确保在每种木糖代谢途径之间可能存在的不同分布中,蛋白质氨基酸中都有可测量且独特的 (13)C 标记分布。与体内结果的比较得出结论,恶臭假单胞菌 VLB120 通过魏默贝格途径代谢木糖。转录组学和生理学研究表明,木糖通过葡萄糖脱氢酶转化为木酸盐。该酶的动力学也负责在存在葡萄糖和木糖的情况下,细胞对葡萄糖作为碳源的偏好。