BackgroundThe halophilic bacterium Chromohalobacter salexigens has been proposed as promising cell factory for the production of the compatible solutes ectoine and hydroxyectoine. This bacterium has evolved metabolic adaptations to efficiently grow under high salt concentrations by accumulating ectoines as compatible solutes. However, metabolic overflow, which is a major drawback for the efficient conversion of biological feedstocks, occurs as a result of metabolic unbalances during growth and ectoines production. Optimal production of ectoines is conditioned by the interplay of carbon and nitrogen metabolisms. In this work, we set out to determine how nitrogen supply affects the production of ectoines.Results
Chromohalobacter salexigens was challenged to grow in media with unbalanced carbon/nitrogen ratio. In C. salexigens, overflow metabolism and ectoines production are a function of medium composition. At low ammonium conditions, the growth rate decreased importantly, up to 80%. Shifts in overflow metabolism were observed when changing the C/N ratio in the culture medium. 13C-NMR analysis of ectoines labelling revealed a high metabolic rigidity, with almost constant flux ratios in all conditions assayed. Unbalanced C/N ratio led to pyruvate accumulation, especially upon N-limitation. Analysis of an ect
− mutant demonstrated the link between metabolic overflow and ectoine biosynthesis. Under non ectoine synthesizing conditions, glucose uptake and metabolic overflow decreased importantly. Finally, in fed-batch cultures, biomass yield was affected by the feeding scheme chosen. High growth (up to 42.4 g L−1) and volumetric ectoine yields (up to 4.21 g L−1) were obtained by minimizing metabolite overflow and nutrient accumulation in high density cultures in a low nitrogen fed-batch culture. Moreover, the yield coefficient calculated for the transformation of glucose into biomass was 30% higher in fed-batch than in the batch culture, demonstrating that the metabolic efficiency of C. salexigens can be improved by careful design of culture feeding schemes.ConclusionsMetabolic shifts observed at low ammonium concentrations were explained by a shift in the energy required for nitrogen assimilation. Carbon-limited fed-batch cultures with reduced ammonium supply were the best conditions for cultivation of C. salexigens, supporting high density growth and maintaining high ectoines production.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-017-0643-7) contains supplementary material, which is available to authorized users.
In recent years, human urine has been successfully used as an electrolyte and organic substrate in bioelectrochemical systems (BESs) mainly due of its unique properties. Urine contains organic compounds that can be utilised as a fuel for energy recovery in microbial fuel cells (MFCs) and it has high nutrient concentrations including nitrogen and phosphorous that can be concentrated and recovered in microbial electrosynthesis cells and microbial concentration cells. Moreover, human urine has high solution conductivity, which reduces the ohmic losses of these systems, improving BES output. This review describes the most recent advances in BESs utilising urine. Properties of neat human urine used in state‐of‐the‐art MFCs are described from basic to pilot‐scale and real implementation. Utilisation of urine in other bioelectrochemical systems for nutrient recovery is also discussed including proofs of concept to scale up systems.
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