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首页> 外文期刊>Bioprocess and Biosystems Engineering >Oxygen transfer as a tool for fine-tuning recombinant protein production by Pichia pastoris under glyceraldehyde-3-phosphate dehydrogenase promoter
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Oxygen transfer as a tool for fine-tuning recombinant protein production by Pichia pastoris under glyceraldehyde-3-phosphate dehydrogenase promoter

机译：氧转移作为微调毕赤酵母在3磷酸甘油醛脱氢酶启动子下重组蛋白生产的工具

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摘要

Effects of oxygen transfer on recombinant protein production by Pichia pastoris under glyceraldehyde- 3-phosphate dehydrogenase promoter were investigated. Recombinant glucose isomerase was chosen as the model protein. Two groups of oxygen transfer strategies were applied, one of which was based on constant oxygen transfer rate where aeration rate was Q(O)/V = 3 and 10 vvm, and agitation rate was N = 900 min(-1); while the other one was based on constant dissolved oxygen concentrations, C-DO = 5, 10, 15, 20 and 40 % in the fermentation broth, by using predetermined exponential glucose feeding with mu(o) = 0.15 h(-1). The highest cell concentration was obtained as 44 g L-1 at t = 9 h of the glucose fed-batch phase at C-DO = 20 % operation while the highest volumetric and specific enzyme activities were obtained as 4440 U L-1 and 126 U g(-1) cell, respectively at C-DO = 15 % operation. Investigation of specific enzyme activities revealed that keeping C-DO at 15 % was more advantageous with an expense of relatively higher byproduct formation and lower specific cell growth rate. For this strategy, the highest oxygen transfer coefficient and oxygen uptake rate were K(L)a = 0.045 s(-1) and OUR = 8.91 mmol m(-3) s(-1), respectively.
机译：研究了在甘油醛3-磷酸脱氢酶启动子作用下，氧转移对巴斯德毕赤酵母重组蛋白生产的影响。选择重组葡萄糖异构酶作为模型蛋白。应用了两组氧气转移策略，其中一组基于恒定的氧气转移速率，其中曝气速率为Q（O）/ V = 3和10 vvm，搅拌速率为N = 900 min（-1）。而另一种是基于恒定的溶解氧浓度，通过使用mu（o）= 0.15 h（-1）的预定指数葡萄糖补料，发酵液中C-DO = 5、10、15、20和40％。在C-DO = 20％操作时，在葡萄糖补料分批阶段的t = 9 h时获得的最高细胞浓度为44 g L-1，而在4440 U L-1和126时获得了最高的体积和比活度U g（-1）电池，分别在C-DO = 15％的操作下。对特定酶活性的研究表明，将C-DO保持在15％更为有利，但要以相对较高的副产物形成和较低的比细胞生长速率为代价。对于此策略，最高的氧气传输系数和氧气吸收率分别为K（L）a = 0.045 s（-1）和OUR = 8.91 mmol m（-3）s（-1）。

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