Growth inhibition of S. cerevisiae, B. subtilis, and E. coli by lignocellulosic and fermentation products

This paper describes the effect of several inhibiting components on three potential hosts for the bio-based production of methyl propionate, namely, wild-type Escherichia coli and Bacillus subtilis, and evolved Saccharomyces cerevisiae IMS0351. The inhibition by the lignocellulose-derived products 5-hydroxymethyl-2-furaldehyde, vanillin, and syringaldehyde and the fermentation products 2-butanol, 2-butanone, methyl propionate, and ethyl acetate has been assessed for these strains in defined medium. Multiple screenings were performed using small-scale cultures in both shake flasks and microtiter plates. Technical drawbacks revealed the limited applicability of the latter in this study. The microbial growth was characterized by means of a lag-time model, and the inhibitory thresholds were determined using product-inhibition models. The lignocellulose-derived products were found to be highly inhibitory, and none of the strains could grow in the presence of 2.0 g L−1 of product. From the fermentation products tested, methyl propionate had the most severe impact resulting in complete inhibition of all the strains when exposed to concentrations in the range of 12–18 g L−1. In general, S. cerevisiae and B. subtilis were comparatively more tolerant than E. coli to all the fermentation products, despite E. coli’s lower sensitivity towards vanillin. The results suggest that, overall, the strains investigated have good potential to be engineered and further established as hosts for the bio-based production of methyl esters.BT/Bioprocess EngineeringOLD BT/Cell Systems Engineerin

A kinetic modelling study of ethane cracking for optimal ethylene yield

Current generation steam cracking plants are considered to be mature. As a consequence it is becoming more and more important to know whether the underlying mechanistic cracking process offers still scope for further improvements. The fundamental kinetic limits to cracking yields have recently been researched in detail for different feed stocks with a new synthesis reactor model, d-RMix, incorporating a large scale mechanistic reaction scheme, SPYRO® [M.W.M. van Goethem, S. Barendregt, J. Grievink, J.A. Moulijn, P.T.J. Verheijen “Model-based, thermo-physical optimisation for high olefin yield in steam cracking reactors”, Chemical Research and Engineering Developments 88 (2010) 1305–1319]. Mathematical optimization revealed for ethane cracking a maximum ethylene yield of about 67 wt%. with a linear-concave optimal temperature profile along the reaction coordinate with a maximum temperature between 1200 and 1300 K. Further mechanistic analysis of these results showed that the linear-concave shape not only suppresses the successive dehydrogenation and condensation reactions of ethylene, but mainly reduces the role of the ethane initiation reaction to form two methyl radicals

Microfluidic Platform with Serpentine Geometry Providing Chaotic Mixing in Induction Time Experiments

We present a droplet microfluidic platform mixing the contents of the droplet chaotically in microfluidic induction time measurements, a promising method for quantifying nucleation kinetics with minute amounts of solute. The nucleation kinetics of aqueous potassium chloride droplets dispersed in mineral oil without surfactants is quantified in the presence and absence of chaotic mixing. We demonstrate the ability of the proposed platform to dictate droplet size, to provide a homogeneous temperature distribution, and to chaotically mix the droplet contents. Chaotic mixing in induction time measurements is facilitated by the motion of droplets through serpentine micromixer bends, while the extent of mixing is controlled by how much droplets move. Different nucleation kinetics are observed in experiments where the droplets are static, mixed, and in motion. We hypothesize that the droplet motion induces formation of a thin-liquid Bretherton film surrounding the droplets. The thin film shields droplets from solid boundaries that are more efficient heteronucleant surfaces compared to liquid-liquid interfaces. We observed that repeated microfluidic induction time measurements, particularly with moving droplets, produce significantly distinct cumulative nucleation probability curves, indicating that the measured nucleation kinetics depend strongly on the details of the experimental procedure, which we discuss in detail. Finally, we compare the microfluidic experiments to well-mixed, milliliter volume, turbidity-based measurements in the context of classic nucleation theory. ChemE/Transport PhenomenaApplied SciencesBT/Design and Engineering EducationComplex Fluid Processin

Experimental design for evaluating WWTP data by linear mass balances

A stepwise experimental design procedure to obtain reliable data from wastewater treatment plants (WWTPs) was developed. The proposed procedure aims at determining sets of additional measurements (besides available ones) that guarantee the identifiability of key process variables, which means that their value can be calculated from other, measured variables, based on available constraints in the form of linear mass balances. Among all solutions, i.e. all possible sets of additional measurements allowing the identifiability of all key process variables, the optimal solutions were found taking into account two objectives, namely the accuracy of the identified key variables and the cost of additional measurements. The results of this multi-objective optimization problem were represented in a Pareto-optimal front. The presented procedure was applied to a full-scale WWTP. Detailed analysis of the relation between measurements allowed the determination of groups of overlapping mass balances. Adding measured variables could only serve in identifying key variables that appear in the same group of mass balances. Besides, the application of the experimental design procedure to these individual groups significantly reduced the computational effort in evaluating available measurements and planning additional monitoring campaigns. The proposed procedure is straightforward and can be applied to other WWTPs with or without prior data collection.Accepted Author ManuscriptBT/Design and Engineering EducationBT/Environmental Biotechnolog

Application of data reconciliation to a dynamically operated wastewater treatment process with off-gas measurements

This study deals with the application of data reconciliation to wastewater treatment processes which are subject to dynamic conditions and therefore do not reach a steady-state behaviour sensu stricto. The SHARON partial nitritation process, which is operated cyclically with alternating aerated and anoxic periods, is studied as an example. The collected data long-term dynamic data set was split up into data subsets corresponding with different pseudo-steady-state operations, which allowed a better gross error detection. Mass balances were set up taking into account off-gas measurements besides liquid phase measurements and including kinetic relations between measurements based on the biological conversions in the reactor. As a result, a higher number of variables could be reconciled, more key variables could be identified, and gross error detection was facilitated. In order to draw conclusions on the process performance in a shorter period of operation, e.g., on the N2O emission factor, the average value of the whole data set should be used with caution. The strong dependence of infiltrated air on the aeration regime and gross error in grab sampling (magnitude of 20%) had a substantial impact on calculating N2O emission. It is recommended that the process performance indicators are derived and checked separately for steady state data subsets to guarantee reliable outcomes.Green Open Access added to TU Delft Institutional Repository 'You share, we take care!' - Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public.BT/Design and Engineering EducationBT/Environmental Biotechnolog

Extreme calorie restriction in yeast retentostats induces uniform non-quiescent growth arrest

Non-dividing Saccharomyces cerevisiae cultures are highly relevant for fundamental and applied studies. However, cultivation conditions in which non-dividing cells retain substantial metabolic activity are lacking. Unlike stationary-phase (SP) batch cultures, the current experimental paradigm for non-dividing yeast cultures, cultivation under extreme calorie restriction (ECR) in retentostat enables non-dividing yeast cells to retain substantial metabolic activity and to prevent rapid cellular deterioration. Distribution of F-actin structures and single-cell copy numbers of specific transcripts revealed that cultivation under ECR yields highly homogeneous cultures, in contrast to SP cultures that differentiate into quiescent and non-quiescent subpopulations. Combined with previous physiological studies, these results indicate that yeast cells subjected to ECR survive in an extended G1 phase. This study demonstrates that yeast cells exposed to ECR differ from carbon-starved cells and offer a promising experimental model for studying non-dividing, metabolically active, and robust eukaryotic cells