Harms H, Zehnder A J
Department of Microbiology, Wageningen Agricultural University, The Netherlands.
Appl Environ Microbiol. 1994 Aug;60(8):2736-45. doi: 10.1128/aem.60.8.2736-2745.1994.
Dibenzofuran uptake-associated kinetic parameters of suspended and attached Sphingomonas sp. strain HH19k cells were compared. The suspended cells were studied in a batch system, whereas glass beads in percolated columns were used as the solid support for attached cells. The maximum specific activities of cells in the two systems were the same. The apparent half-maximum uptake rate-associated concentrations (Kt') of attached cells, however, were considerably greater than those of suspended cells and depended on cell density and on percolation velocity. A mathematical model was developed to explain the observed differences in terms of substrate transport to the cells. This model was based on the assumptions that the intrinsic half-maximum uptake rate-associated concentration (Kt) was unchanged and that deviations of Kt' from Kt resulted from the stereometry and the hydrodynamics around the cells. Our calculations showed that (i) diffusion to suspended cells and to single attached cells is efficient and therefore only slightly affects Kt'; (ii) diffusion to cells located on crowded surfaces is considerably lower than that to single attached cells and greatly increases Kt', which depends on the cell density; (iii) the convective-diffusive transport to attached cells that occurs in a percolated column is influenced by the liquid flow and results in dependency of Kt' on the flow rate; and (iv) higher specific affinity of cells correlates with higher susceptibility to diffusion limitation. Properties of the experimental system which limited quantitative proof of exclusively transport-controlled variations of Kt' are discussed.
比较了悬浮和附着的鞘氨醇单胞菌属菌株HH19k细胞对二苯并呋喃摄取相关的动力学参数。悬浮细胞在分批系统中进行研究,而在渗滤柱中的玻璃珠用作附着细胞的固体支持物。两个系统中细胞的最大比活性相同。然而,附着细胞的表观半最大摄取速率相关浓度(Kt')远高于悬浮细胞,并且取决于细胞密度和渗滤速度。建立了一个数学模型来解释观察到的底物向细胞转运方面的差异。该模型基于以下假设:内在半最大摄取速率相关浓度(Kt)不变,并且Kt'与Kt的偏差是由细胞周围的立体测量和流体动力学引起的。我们的计算表明:(i)向悬浮细胞和单个附着细胞的扩散是有效的,因此仅对Kt'有轻微影响;(ii)向位于拥挤表面的细胞的扩散远低于向单个附着细胞的扩散,并且大大增加了Kt',这取决于细胞密度;(iii)在渗滤柱中发生的向附着细胞的对流扩散传输受液体流动影响,并导致Kt'对流速的依赖性;(iv)细胞的较高比亲和力与对扩散限制的较高敏感性相关。讨论了限制对Kt'仅由转运控制的变化进行定量证明的实验系统特性。