Improved vanillin production in baker's yeast through in silico design

<p>Abstract</p> <p>Background</p> <p>Vanillin is one of the most widely used flavouring agents, originally obtained from cured seed pods of the vanilla orchid <it>Vanilla planifolia</it>. Currently vanillin is mostly produced <it>via </it>chemical synthesis. A <it>de novo </it>synthetic pathway for heterologous vanillin production from glucose has recently been implemented in baker's yeast, <it>Saccharamyces cerevisiae</it>. In this study we aimed at engineering this vanillin cell factory towards improved productivity and thereby at developing an attractive alternative to chemical synthesis.</p> <p>Results</p> <p>Expression of a glycosyltransferase from <it>Arabidopsis thaliana </it>in the vanillin producing <it>S. cerevisiae </it>strain served to decrease product toxicity. An <it>in silico </it>metabolic engineering strategy of this vanillin glucoside producing strain was designed using a set of stoichiometric modelling tools applied to the yeast genome-scale metabolic network. Two targets (<it>PDC1 </it>and <it>GDH1</it>) were selected for experimental verification resulting in four engineered strains. Three of the mutants showed up to 1.5 fold higher vanillin β-D-glucoside yield in batch mode, while continuous culture of the <it>Δpdc1 </it>mutant showed a 2-fold productivity improvement. This mutant presented a 5-fold improvement in free vanillin production compared to the previous work on <it>de novo </it>vanillin biosynthesis in baker's yeast.</p> <p>Conclusion</p> <p>Use of constraints corresponding to different physiological states was found to greatly influence the target predictions given minimization of metabolic adjustment (MOMA) as biological objective function. <it>In vivo </it>verification of the targets, selected based on their predicted metabolic adjustment, successfully led to overproducing strains. Overall, we propose and demonstrate a framework for <it>in silico </it>design and target selection for improving microbial cell factories.</p

Workplace sexual harassment and depressive symptoms:A cross-sectional multilevel analysis comparing harassment from clients or customers to harassment from other employees amongst 7603 Danish employees from 1041 organizations

Abstract Background Previous research has reported that sexual harassment can lead to reduced mental health. Few studies have focused on sexual harassment conducted by clients or customers, which might occur in person-related occupations such as eldercare work, social work or customer service work. This study examined the cross-sectional association between sexual harassment by clients or customers and depressive symptoms. We also examined if this association was different compared to sexual harassment conducted by a colleague, supervisor or subordinate. Further, we investigated if psychosocial workplace initiatives modified the association between sexual harassment by clients or customers and level of depressive symptoms. Methods We used data from the Work Environment and Health in Denmark cohort study (WEHD) and the Work Environment Activities in Danish Workplaces Study (WEADW) collected in 2012. WEHD is based on a random sample of employed individuals aged 18–64. In WEADW, organizational supervisors or employee representatives provided information on workplace characteristics. By combining WEHD and WEADW we included self-reported information on working conditions and health from 7603 employees and supervisors in 1041 organizations within 5 occupations. Data were analyzed using multilevel regression and analyses adjusted for gender, age, occupation and socioeconomic position. Results Exposure to workplace sexual harassment from clients or customers was statistically significantly associated with a higher level of depressive symptoms (2.05; 95% CI: 0.98–3.12) compared to no exposure. Employees harassed by colleagues, supervisors or subordinates had a higher mean level of depressive symptoms (2.45; 95% CI: 0.57–4.34) than employees harassed by clients or customers. We observed no statistically significant interactions between harassment from clients and customers and any of the examined psychosocial workplace initiatives (all p > 0.05). Conclusions The association between sexual harassment and depressive symptoms differed for employees harassed by clients or customers and those harassed by colleagues, supervisors or subordinates. The results underline the importance of investigating sexual harassment from clients or customers and sexual harassment by colleagues, supervisors or subordinates as distinct types of harassment. We found no modification of the association between sexual harassment by clients or customers and depressive symptoms by any of the examined psychosocial workplace initiatives

Viscous stabilization of 2D drainage displacements with trapping

We investigate the stabilization mechanisms due to viscous forces in the invasion front during drainage displacement in two-dimensional porous media using a network simulator. We find that in horizontal displacement the capillary pressure difference between two different points along the front varies almost linearly as function of height separation in the direction of the displacement. The numerical result supports arguments taking into account the loopless displacement pattern where nonwetting fluid flow in separate strands (paths). As a consequence, we show that existing theories developed for viscous stabilization, are not compatible with drainage when loopless strands dominate the displacement process.Comment: The manuscript has been substantially revised. Accepted in Phys. Rev. Let