Rönsch Hendrik, Krämer Reinhard, Morbach Susanne
Institut für Biochemie der Universität zu Köln, D-50674 Köln, Germany.
J Biotechnol. 2003 Sep 4;104(1-3):87-97. doi: 10.1016/s0168-1656(03)00166-4.
The response of the L-lysine producing Corynebacterium glutamicum strain MH20-22B to osmotic stress was studied in batch cultures. To mimic the conditions during a fermentation process the long term adaptation of cells subjected to a constant osmotic stress between 1.0 and 2.5 osM was investigated. Cytoplasmic water content and volume of C. glutamicum cells were found to depend on growth phase, extent of osmotic stress and availability of betaine. The maximal cytoplasmic volumes, which were highest at maximal growth rate, were linearily related to osmotic stress, whereas in stationary cells no active volume regulation was observed. Under severe osmotic stress proline was the prominent compatible solute in growing cells. Uptake of betaine, if available in the medium, reduced the concentration of proline from 750 to 300 mM, indicating that uptake of compatible solutes is preferred to synthesis. Furthermore, betaine was shown to have a higher efficiency to counteract osmotic stress, since the overall concentration of compatible solutes was lower in the presence of betaine. Under severe osmotic stress, the addition of betaine shifted L-lysine production in MH20-22B to earlier fermentation times and increased both product concentration and yield in these phases, but did not improve the final L-lysine yield.
在分批培养中研究了产L-赖氨酸的谷氨酸棒杆菌菌株MH20-22B对渗透胁迫的响应。为模拟发酵过程中的条件,研究了细胞在1.0至2.5 osM恒定渗透胁迫下的长期适应性。发现谷氨酸棒杆菌细胞的细胞质含水量和体积取决于生长阶段、渗透胁迫程度和甜菜碱的可用性。最大细胞质体积在最大生长速率时最高,与渗透胁迫呈线性关系,而在静止细胞中未观察到主动体积调节。在严重渗透胁迫下,脯氨酸是生长细胞中主要的相容性溶质。如果培养基中有甜菜碱,其摄取会使脯氨酸浓度从750 mM降至300 mM,这表明摄取相容性溶质比合成更受青睐。此外,甜菜碱显示出更高的抵抗渗透胁迫的效率,因为在有甜菜碱存在的情况下,相容性溶质的总浓度较低。在严重渗透胁迫下,添加甜菜碱使MH20-22B中的L-赖氨酸生产提前至发酵前期,并提高了这些阶段的产物浓度和产量,但未提高最终L-赖氨酸产量。