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大肠杆菌在有氧和中温培养条件下葡萄糖利用的呼吸测定评估与建模

Respirometric evaluation and modeling of glucose utilization by Escherichia coli under aerobic and mesophilic cultivation conditions.

作者信息

Insel G, Celikyilmaz G, Ucisik-Akkaya E, Yesiladali K, Cakar Z P, Tamerler C, Orhon D

机构信息

Department of Environmental Engineering, Istanbul Technical University, Maslak, 34469, Istanbul, Turkey.

出版信息

Biotechnol Bioeng. 2007 Jan 1;96(1):94-105. doi: 10.1002/bit.21163.

Abstract

The study presents a mechanistic model for the evaluation of glucose utilization by Escherichia coli under aerobic and mesophilic growth conditions. In the first step, the experimental data was derived from batch respirometric experiments conducted at 37 degrees C, using two different initial substrate to microorganism (S(0)/X(0)) ratios of 15.0 and 1.3 mgCOD/mgSS. Acetate generation, glycogen formation and oxygen uptake rate profile were monitored together with glucose uptake and biomass increase throughout the experiments. The oxygen uptake rate (OUR) exhibited a typical profile accounting for growth on glucose, acetate and glycogen. No acetate formation (overflow) was detected at low initial S(0)/X(0) ratio. In the second step, the effect of culture history developed under long-term growth limiting conditions on the kinetics of glucose utilization by the same culture was evaluated in a sequencing batch reactor (SBR). The system was operated at cyclic steady state with a constant mean cell residence time of 5 days. The kinetic response of E.coli culture was followed by similar measurements within a complete cycle. Model calibration for the SBR system showed that E. coli culture regulated its growth metabolism by decreasing the maximum growth rate (lower microH) together with an increase of substrate affinity (lower K(S)) as compared to uncontrolled growth conditions. The continuous low rate operation of SBR system induced a significant biochemical substrate storage capability as glycogen in parallel to growth, which persisted throughout the operation. The acetate overflow was observed again as an important mechanism to be accounted for in the evaluation of process kinetics.

摘要

该研究提出了一个机理模型,用于评估嗜温好氧生长条件下大肠杆菌对葡萄糖的利用情况。第一步,实验数据源自于在37℃下进行的间歇呼吸测定实验,使用了两种不同的初始底物与微生物(S(0)/X(0))比率,分别为15.0和1.3 mgCOD/mgSS。在整个实验过程中,监测了乙酸盐生成、糖原形成和氧摄取率曲线,同时监测了葡萄糖摄取和生物量增加情况。氧摄取率(OUR)呈现出典型的曲线,反映了在葡萄糖、乙酸盐和糖原上的生长情况。在低初始S(0)/X(0)比率下未检测到乙酸盐形成(溢流)。第二步,在序批式反应器(SBR)中评估了在长期生长限制条件下形成的培养历史对同一培养物葡萄糖利用动力学的影响。该系统在循环稳态下运行,平均细胞停留时间恒定为5天。在一个完整周期内通过类似测量跟踪大肠杆菌培养物的动力学响应。SBR系统的模型校准表明,与非控制生长条件相比,大肠杆菌培养物通过降低最大生长速率(较低的μmax)以及增加底物亲和力(较低的K(S))来调节其生长代谢。SBR系统的连续低速率运行诱导了一种显著的生化底物储存能力,即与生长并行的糖原储存能力,这种能力在整个运行过程中持续存在。再次观察到乙酸盐溢流是评估过程动力学时需要考虑的一个重要机制。

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