High cell density cultivation combined with high specific enzyme activity: Cultivation protocol for the production of an amine transaminase from Bacillus megaterium in E. coli.

巨芽孢杆菌 转氨酶 生物化学 化学 产量(工程) 生物量(生态学) 生物技术 食品科学 生物 细菌 材料科学 遗传学 农学 冶金
作者
Berit Rothkranz,Matthias Rieb,Evelin Lisa Unrau,Ilona Frindi-Wosch,Johannes Hemmerich,Torsten Sehl,Dörte Rother
出处
期刊:ChemBioChem [Wiley]
标识
DOI:10.1002/cbic.202400006
摘要

High cell density cultivation is an established method for the production of various industrially important products such as recombinant proteins. However, these protocols are often not suitable for biocatalytic processes as often the focus lies on biomass production rather than high specific activities of the enzyme. In contrast, a range of shake flask protocols are well known with high specific activities but rather low cell densities. To overcome this gap, we established a tailor‐made fed‐batch protocol combining both aspects: high cell density and high specific activities of heterologously produced enzyme. Using the example of an industrially relevant amine transaminase from Bacillus megaterium, we describe a strategy to optimize the cultivation yield based on the feed rate, IPTG concentration and post‐induction temperature. By adjusting these key parameters, we were able to increase the specific activity by 2.6‐fold and the wet cell weight by even 17‐fold compared to shake flasks. Finally, we were able to verify our established protocol by transferring it to another operator. With that, our optimization strategy can serve as a template for the production of high titers of active heterologously produced enzymes and might enable the availability of these catalysts for upscaling of biocatalytic processes.
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