Lactose autoinduction with enzymatic glucose release: Characterization of the cultivation system in bioreactor

详细信息    查看全文

• 作者：Sonja Mayer ; Stefan Junne ; Kaisa Ukkonen ; Julia Glazyrina ; Florian Glauche ; Peter Neubauer ; Antti Vasala
• 关键词：Lactose autoinduction ; Enzymatic glucose delivery ; Fed-batch ; Bioreactor
• 刊名：Protein Expression and Purification
• 出版年：February, 2014
• 年：2014
• 卷：94
• 期：Complete
• 页码：67-72
• 全文大小：618 K

文摘

The lactose autoinduction system for recombinant protein production was combined with enzymatic glucose release as a method to provide a constant feed of glucose instead of using glycerol as a carbon substrate. Bioreactor cultivation confirmed that the slow glucose feed does not prevent the induction by lactose. HPLC studies showed that with successful recombinant protein production only a very low amount of lactose was metabolized during glucose-limited fed-batch conditions by the Escherichia coli strain BL21(DE3)pLysS in well-aerated conditions, which are problematic for glycerol-based autoinduction systems. We propose that slow enzymatic glucose feed does not cause a full activation of the lactose operon. However recombinant PDI-A protein (A-domain of human disulfide isomerase) was steadily produced until the end of the cultivation. The results of the cultivations confirmed our earlier observations with shaken cultures showing that lactose autoinduction cultures based on enzymatic glucose feed have good scalability, and that this system can be applied also to bioreactor cultivations.