Delft University of Technology, Department of Biochemical Engineering, 2628 BC Delft, The Netherlands.
Biotechnol Bioeng. 1990 Nov;36(9):931-9. doi: 10.1002/bit.260360908.
An oxygen microsensor in combination with mathematical modeling was used to determine the behavior of immobilized Thiosphaera pantotropha. This organism can convert ammonia completely to nitrogen gas under aerobic conditions (coupled nitrification/denitrification) and denitrifies nitrate at highest rates under anaerobic conditions. Immobilization of T. pantotropha can result in aerobic and anaerobic zones inside the biocatalyst particle which will be advantageous for the conversion of ammonia and nitrate from wastewater. However, information of the effects of immobilization on the physiology of T. pantotropha is necessary for the development of such a system. This article gives the extension of a model developed to describe the behavior of chemostat cultures of T. pantotropha so that it can be used for immobilized cells. The original model was based on metabolic reaction equations. Kinetic and diffusion equations have now been added. Experimental verification was carried out using a stirred tank reactor and a Kluyver flask. After immobilization in agarose, the cells were grown in the particles under continuous culture conditions for 3 days. After 24 h the oxygen penetration depth showed a constant value of 100 microm, indicating that a steady state was reached. Scanning electron micrographs showed that large colonies of cells were present in this 100-microm aerobic layer.From the dynamics of the start-up phase, several parameters were determined from measurements of the oxygen concentration profiles made every few hours. The profiles simulated by the model were fitted to the measured data. The average value for the maximum specific growth rate was 0.52 h(-1), and the maximum oxygen conversion rate was 1.0 mol Cmol(-1) h(-1). The maximum specific acetate uptake rate was 2.0 mol Cmol(-1) h(-1), and the Monod constant for acetate was 2.9 x 10(-2) mol m(-3). The maximum specific nitrification rate was 0.58 x 10(-1) mol Cmol(-1) h(-1), and the amount of oxygen necessary for nitrification was 11% of the total oxygen uptake rate. Most of the kinetic parameters determined for the immobilized cells were in good agreement with those for the suspended cells. Only the maximum specific growth rate was significantly higher, and the maximum specific nitrification rate was some what lower than for suspended cells. The experimental results clearly show that an oxygen microsensor, in combination with mathematical modeling, can successfully be used to elucidate the kinetic behavior of immobilized, oxygen-consuming, cells.
采用氧微传感器与数学模型相结合的方法来测定固定化脱硫弧菌的行为。该菌在好氧条件下能将氨完全转化为氮气(偶联硝化/反硝化),在厌氧条件下以最高速率反硝化硝酸盐。T. pantotropha 的固定化会导致生物催化剂颗粒内出现好氧区和厌氧区,这有利于从废水中转化氨和硝酸盐。然而,为了开发这种系统,需要了解固定化对 T. pantotropha 生理学的影响。本文给出了为描述 T. pantotropha 恒化培养物行为而开发的模型的扩展,以便能够将其用于固定化细胞。原始模型基于代谢反应方程。现在已经添加了动力学和扩散方程。使用搅拌槽反应器和 Kluyver 瓶进行了实验验证。在琼脂糖中固定化后,细胞在连续培养条件下在颗粒中生长了 3 天。24 小时后,氧渗透深度显示出 100 微米的恒定值,表明达到了稳定状态。扫描电子显微镜显示,在这个 100 微米的好氧层中有大的细胞菌落。从启动阶段的动力学来看,通过每隔几个小时测量一次氧浓度分布,可以确定几个参数。通过模型模拟的分布与测量数据相拟合。最大比生长速率的平均值为 0.52 h(-1),最大氧转化速率为 1.0 mol Cmol(-1) h(-1)。最大比乙酸吸收速率为 2.0 mol Cmol(-1) h(-1),乙酸的 Monod 常数为 2.9 x 10(-2) mol m(-3)。最大硝化速率为 0.58 x 10(-1) mol Cmol(-1) h(-1),硝化所需的氧量为总耗氧量的 11%。为固定化细胞确定的大多数动力学参数与悬浮细胞的参数非常吻合。只有最大比生长速率显著较高,最大硝化速率略低于悬浮细胞。实验结果清楚地表明,氧微传感器与数学建模相结合可成功用于阐明耗氧固定化细胞的动力学行为。