Institute of Biochemical Engineering, Technical University of Munich, Boltzmannstr. 15, D-85748, Garching, Germany.
Chair of Microbiology, Technical University of Munich, Emil-Ramann-Str. 4, D-85354, Freising, Germany.
Appl Microbiol Biotechnol. 2019 Jun;103(11):4393-4404. doi: 10.1007/s00253-019-09824-w. Epub 2019 Apr 18.
Many ketoses or organic acids can be produced by membrane-associated oxidation with Gluconobacter oxydans. In this study, the oxidation of meso-erythritol to L-erythrulose was investigated with the strain G. oxydans 621HΔupp BP.8, a multideletion strain lacking the genes for eight membrane-bound dehydrogenases. First batch biotransformations with growing cells showed re-consumption of L-erythrulose by G. oxydans 621HΔupp BP.8 in contrast to resting cells. The batch biotransformation with 2.8 g L resting cells of G. oxydans 621HΔupp BP.8 in a DO-controlled stirred-tank bioreactor resulted in 242 g L L-erythrulose with a product yield of 99% (w/w) and a space-time yield of 10 g L h. Reaction engineering studies showed substrate excess inhibition as well as product inhibition of G. oxydans 621HΔupp BP.8 in batch biotransformations. In order to overcome substrate inhibition, a continuous membrane bioreactor with full cell retention was applied for meso-erythritol oxidation with resting cells of G. oxydans 621HΔupp BP.8. At a mean hydraulic residence time of 2 h, a space-time yield of 27 g L h L-erythrulose was achieved without changing the product yield of 99% (w/w) resulting in a cell-specific product yield of up to 4.4 g g in the steady state. The product concentration (54 g L L-erythrulose) was reduced in the continuous biotransformation process compared with the batch process to avoid product inhibition.
许多酮糖或有机酸可以通过与氧化葡萄糖酸杆菌的膜相关氧化产生。在这项研究中,使用多缺失菌株 G. oxydans 621HΔupp BP.8(缺乏 8 个膜结合脱氢酶基因)对 meso-erythritol 氧化为 L-erythrulose 进行了研究。与静止细胞相比,生长细胞的第一批分批转化显示 G. oxydans 621HΔupp BP.8 重新消耗 L-erythrulose。在 DO 控制的搅拌釜生物反应器中,用 2.8 g L 的静止细胞进行分批转化,得到 242 g L 的 L-erythrulose,产物收率为 99%(w/w),时空产率为 10 g L h。反应工程研究表明,在分批转化中,底物过量抑制以及产物抑制 G. oxydans 621HΔupp BP.8。为了克服底物抑制,使用全细胞保留的连续膜生物反应器对 G. oxydans 621HΔupp BP.8 的静止细胞进行了 meso-erythritol 氧化。在平均水力停留时间为 2 h 的情况下,实现了 27 g L h L-erythrulose 的时空产率,而产物收率保持在 99%(w/w)不变,从而在稳态下达到高达 4.4 g g 的细胞特异性产物产率。与分批转化相比,连续生物转化过程中的产物浓度(54 g L L-erythrulose)降低,以避免产物抑制。