RESUMEN
Engineering microbial cells for the commercial production of biomolecules and biochemicals requires understanding how cells respond to dynamically changing substrate (feast-famine) conditions in industrial-scale bioreactors. Scale-down methods that oscillate substrate are commonly applied to predict the industrial-scale behavior of microbes. We followed a compartment modeling approach to design a scale-down method based on the simulation of an industrial-scale bioreactor. This study uses high cell-density scale-down experiments to investigate Escherichia coli knockout strains of five major glucose-sensitive transcription factors (Cra, Crp, FliA, PrpR, and RpoS) to study their regulatory role during glucose oscillations. RNA-sequencing analysis revealed that the glucose oscillations caused the down-regulation of several stress-related functions in E. coli. An in-depth analysis of strain physiology and transcriptome revealed a distinct phenotype of the strains tested under glucose oscillations. Specifically, the knockout strains of Cra, Crp, and RpoS resulted in a more sensitive transcriptional response than the control strain, while the knockouts of FliA and PrpR responded less severely. These findings imply that the regulation orchestrated by Cra, Crp, and RpoS may be essential for robust E. coli production strains. In contrast, the regulation by FliA and PrpR may be undesirable for temporal oscillations in glucose availability.
RESUMEN
In the present work, tomato peels were pre-treated using combination of ultrasound and enzyme co-immobilized amino-functionalized magnetic nanoparticles (AMNPs) for the efficient release of lycopene. To achieve maximum activity of enzymes in the co-immobilized form, optimization of several parameters were carried out. Moreover, the influence of ultrasound and enzyme co-immobilized magnetic nanoparticles on lycopene release was studied. Maximum lycopene release was obtained at 3% (w/w) enzyme co-immobilized AMNPs, pH 5.0, temperature of 50⯰C, at 10â¯W ultrasound power and 20â¯min incubation time. After enzymatic pre-treatment, lycopene from the pre-treated mixture was extracted and separated using tri-solvent extraction method. Maximum recovery of lycopene using solvent extraction was obtained at 50⯰C, 90â¯min of incubation time and agitation speed of 150â¯rpm. The presence of lycopene in the extract was confirmed by FT-IR, UV-vis spectroscopy and HPLC analysis. The co-immobilized bio-catalyst showed excellent reusability giving more than 50% lycopene yield even after 6th cycles of reuse.