Scale-up bioprocess development forproduction of the antibiotic valinomycin in Escherichiacoli based on consistent fed-batch cultivations

Microbial Cell Factories

Research

Jennifer Jaitzig¹  Ping Lu¹  Peter Neubauer¹  Jian Li²  Roderich D Süssmuth³ 

[1] Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstraße 76, ACK24, 13355, Berlin, Germany;Chair of Bioprocess Engineering, Department of Biotechnology, Technische Universität Berlin, Ackerstraße 76, ACK24, 13355, Berlin, Germany;Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Road, 60208, Evanston, IL, USA;Department of Chemistry, Technische Universität Berlin, Straße des 17. Juni 124, 10623, Berlin, Germany;

关键词: Valinomycin;    Synthetic biology;    Bioprocess development;    Scale-up;    Fed-batch;    Escherichia coli;   

DOI  :  10.1186/s12934-015-0272-y

 received in 2015-02-05, accepted in 2015-05-08,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundHeterologous production of natural products in Escherichia coli has emerged as an attractivestrategy to obtain molecules of interest. Although technically feasible most ofthem are still constrained to laboratory scale production. Therefore, it isnecessary to develop reasonable scale-up strategies for bioprocesses aiming atthe overproduction of targeted natural products under industrial scaleconditions. To this end, we used the production of the antibiotic valinomycin inE. coli as a model system for scalablebioprocess development based on consistent fed-batch cultivations.ResultsIn this work, the glucose limited fed-batch strategy based on puremineral salt medium was used throughout all scales for valinomycin production.The optimal glucose feed rate was initially detected by the use of abiocatalytically controlled glucose release (EnBase® technology) in parallelcultivations in 24-well plates with continuous monitoring of pH and dissolvedoxygen. These results were confirmed in shake flasks, where the accumulation ofvalinomycin was highest when the specific growth rate decreased below0.1 h−1. This correlation was also observed forhigh cell density fed-batch cultivations in a lab-scale bioreactor. Thebioreactor fermentation produced valinomycin with titers of more than2 mg L−1 based on the feeding of a concentratedglucose solution. Valinomycin production was not affected by oscillatingconditions (i.e. glucose and oxygen) in a scale-down two-compartment reactor,which could mimic similar situations in industrial bioreactors, suggesting thatthe process is very robust and a scaling of the process to a larger industrialscale appears a realistic scenario.ConclusionsValinomycin production was scaled up from mL volumes to 10 L withconsistent use of the fed-batch technology. This work presents a robust andreliable approach for scalable bioprocess development and represents an examplefor the consistent development of a process for a heterologously expressednatural product towards the industrial scale.

【 授权许可】

CC BY   
© Li et al. 2015