Department of Chemical Engineering, Biotechnology and Materials, Centre for Biotechnology and Bioengineering (CeBiB), University of Chile, Beauchef 850, 8370448, Santiago, Chile.
Systems and Synthetic Biology Group, Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, Brisbane, Australia.
Biotechnol Lett. 2019 Jul;41(6-7):779-788. doi: 10.1007/s10529-019-02680-8. Epub 2019 May 7.
Over-express galactokinase (Galk1) in tissue plasminogen activator (tPA) producing CHO cells as a potential strategy to improve cell growth and product synthesis.
tPA producing CHO cells were transfected with the galactokinase (Galk1) gene. CHO-Galk1 cells showed a 39% increase of the specific growth rate in galactose. Moreover, clones were able to use this hexose as their main carbon source to sustain growth contrary to their parental cell line. Metabolic Flux Analysis revealed that the CHO-Galk1 selected clone shows an active metabolism towards biomass and product synthesis, characterized by higher fluxes in the TCA cycle, which is consistent with increased cellular densities and final product concentration.
This cellular engineering strategy, where modifications of key points of alternative carbon sources metabolism lead to an improved metabolism of these sugars, is a starting point towards the generation of new cell lines with reduced lactate synthesis and increased cell growth and productivity.
在组织型纤溶酶原激活物（tPA）产生的 CHO 细胞中过度表达半乳糖激酶（Galk1），作为提高细胞生长和产物合成的潜在策略。
用半乳糖激酶（Galk1）基因转染 tPA 产生的 CHO 细胞。CHO-Galk1 细胞在半乳糖中的比生长速率提高了 39%。此外，与亲本细胞系相比，克隆能够将这种己糖用作主要碳源来维持生长。代谢通量分析表明，CHO-Galk1 选择的克隆显示出对生物量和产物合成的活跃代谢，其特征是 TCA 循环中的通量更高，这与细胞密度的增加和最终产物浓度的增加一致。
这种细胞工程策略，其中对替代碳源代谢的关键点进行修饰导致这些糖的代谢得到改善，是朝着生成新的细胞系迈出的第一步，这些细胞系的乳酸合成减少，细胞生长和生产力提高。
