Lú-Chau T A, Guillán A, Núñez M J, Roca E, Lema J M
Department of Chemical Engineering, Institute of Technology, University of Santiago de Compostela, E-15706 Santiago de Compostela, Spain.
Bioprocess Biosyst Eng. 2004 Apr;26(3):159-63. doi: 10.1007/s00449-003-0344-y. Epub 2004 Feb 18.
Two bioreactor continuous cultures, at anaerobic and aerobic conditions, were carried out using a recombinant Saccharomyces cerevisiae strain that over-expresses the homologous gene EXG1. This recombinant system was used to study the effect of dissolved oxygen concentration on plasmid stability and gene over-expression. Bioreactor cultures were operated at two dilution rates (0.14 and 0.03 h(-1)) to investigate the effect of other process parameters on EXG1 expression. Both cultures suffered severe plasmid instability during the first 16 generations. Segregational plasmid loss rate for the aerobic culture was two-fold that of the anaerobic operation. In spite of this fact, exo-beta-glucanase activity at aerobic conditions was 12-fold that of the anaerobic culture. This maximal activity (30 U ml(-1)) was attained at the lowest dilution rate when biomass reached its greatest value and glucose concentration was zero.
使用过表达同源基因EXG1的重组酿酒酵母菌株,在厌氧和好氧条件下进行了两种生物反应器连续培养。该重组系统用于研究溶解氧浓度对质粒稳定性和基因过表达的影响。生物反应器培养以两种稀释率(0.14和0.03 h(-1))运行,以研究其他工艺参数对EXG1表达的影响。两种培养在最初16代期间均遭受严重的质粒不稳定性。好氧培养的分离质粒丢失率是厌氧操作的两倍。尽管如此,好氧条件下的外切β-葡聚糖酶活性是厌氧培养的12倍。当生物量达到最大值且葡萄糖浓度为零时,在最低稀释率下达到了这种最大活性(30 U ml(-1))。