Optimization of norovirus virus‐like particle production in Pichia pastoris using a real‐time near‐infrared bioprocess monitor

The production of norovirus virus‐like particles (NoV VLPs) displaying NY‐ESO‐1 cancer testis antigen in Pichia pastoris BG11 Mut⁺ has been enhanced through feed‐strategy optimization using a near‐infrared bioprocess monitor (RTBio^® Bioprocess Monitor, ASL Analytical, Inc.), capable of monitoring and controlling the concentrations of glycerol and methanol in real‐time. The production of NoV VLPs displaying NY‐ESO‐1 in P. pastoris has potential as a novel cancer vaccine platform. Optimization of the growth conditions resulted in an almost two‐fold increase in the expression levels in the fermentation supernatant of P. pastoris as compared to the starting conditions. We investigated the effect of methanol concentration, batch phase time, and batch to induction transition on NoV VLP‐NY‐ESO‐1 production. The optimized process included a glycerol transition phase during the first 2 h of induction and a methanol concentration set point of 4 g L^?1 during induction. Utilizing the bioprocess monitor to control the glycerol and methanol concentrations during induction resulted in a maximum NoV VP1‐NY‐ESO‐1 yield of 0.85 g L^?1. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:518–526, 2016     

Toward intensifying design of experiments in upstream bioprocess development: An industrial Escherichia coli feasibility study

In this study, step variations in temperature, pH, and carbon substrate feeding rate were performed within five high cell density Escherichia coli fermentations to assess whether intraexperiment step changes, can principally be used to exploit the process operation space in a design of experiment manner. A dynamic process modeling approach was adopted to determine parameter interactions. A bioreactor model was integrated with an artificial neural network that describes biomass and product formation rates as function of varied fed‐batch fermentation conditions for heterologous protein production. A model reliability measure was introduced to assess in which process region the model can be expected to predict process states accurately. It was found that the model could accurately predict process states of multiple fermentations performed at fixed conditions within the determined validity domain. The results suggest that intraexperimental variations of process conditions could be used to reduce the number of experiments by a factor, which in limit would be equivalent to the number of intraexperimental variations per experiment. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1343–1352, 2016     

Demographic buffering and compensatory recruitment promotes the persistence of disease in a wildlife population

Demographic buffering allows populations to persist by compensating for fluctuations in vital rates, including disease‐induced mortality. Using long‐term data on a badger (Meles meles Linnaeus, 1758) population naturally infected with Mycobacterium bovis, we built an integrated population model to quantify impacts of disease, density and environmental drivers on survival and recruitment. Badgers exhibit a slow life‐history strategy, having high rates of adult survival with low variance, and low but variable rates of recruitment. Recruitment exhibited strong negative density‐dependence, but was not influenced by disease, while adult survival was density independent but declined with increasing prevalence of diseased individuals. Given that reproductive success is not depressed by disease prevalence, density‐dependent recruitment of cubs is likely to compensate for disease‐induced mortality. This combination of slow life history and compensatory recruitment promotes the persistence of a naturally infected badger population and helps to explain the badger's role as a persistent reservoir of M. bovis.     

An indicator framework for assessing the technology aspect of Integrated Lake Basin Management for Lake Malawi Basin

Lake Malawi is one of the African Great Lakes and is well known for its fish biodiversity which is the greatest in the world for freshwaters. The lake basin is a valuable resource to the riparian countries. Activities associated with the derivation of the values offered by the lake basin, however, result in the generation of wastes and pollutants that also impact on the lake basin. Such anthropogenic activities within the lake basin place considerable strain on the lake system and may lead to the degradation of the lake basin. Proper application of technology can effectively supplement command-and-control measures and economic instruments for better lake basin management. Currently, no comprehensive framework exists to assess the management of the lake basin. Such a framework is necessary to guide managers and policy-makers in the formulation of management plans and strategies for the sustainable management and utilization of the lake basin. We propose an indicator framework for assessing the technology aspect of Integrated Lake Basin Management (ILBM) in the context of the Lake Malawi Basin. We further demonstrate applicability of the proposed framework through a study conducted in the Malawian side of the basin. We applied a five point Likert-type scale in our indicator assessment. Overall, our results show that the technology aspect of the management of the lake basin is weak to moderate. This threatens the sustainability of the lake especially in the areas of solid waste management, sanitation services, pollution control and protection of wetlands and lagoons which performed from very low to moderate. The major challenges are weak institutional capacity, weak regulation enforcement, and insufficient resources.     

Proteomic analysis of Rac1 signaling regulation by guanine nucleotide exchange factors

The small GTPase Rac1 is implicated in various cellular processes that are essential for normal cell function. Deregulation of Rac1 signaling has also been linked to a number of diseases, including cancer. The diversity of Rac1 functioning in cells is mainly attributed to its ability to bind to a multitude of downstream effectors following activation by Guanine nucleotide Exchange Factors (GEFs). Despite the identification of a large number of Rac1 binding partners, factors influencing downstream specificity are poorly defined, thus hindering the detailed understanding of both Rac1's normal and pathological functions. In a recent study, we demonstrated a role for 2 Rac-specific GEFs, Tiam1 and P-Rex1, in mediating Rac1 anti- versus pro-migratory effects, respectively. Importantly, via conducting a quantitative proteomic screen, we identified distinct changes in the Rac1 interactome following activation by either GEF, indicating that these opposing effects are mediated through GEF modulation of the Rac1 interactome. Here, we present the full list of identified Rac1 interactors together with functional annotation of the differentially regulated Rac1 binding partners. In light of this data, we also provide additional insights into known and novel signaling cascades that might account for the GEF-mediated Rac1-driven cellular effects.     

Field efficacy of insect pathogen,botanical, and jasmonic acid for the management of wheat midge Sitodiplosis mosellana and the impact on adult parasitoid Macroglenes penetrans populations in spring wheat

The wheat midge, Sitodiplosis mosellana, is a serious pest of wheat worldwide. In North America, management of S. mosellana in spring wheat relies on the timely application of pesticides, based on midge adults levels caught in pheromone traps or seen via field scouting during wheat heading. In this context, biopesticides can be an effective alternative to pesticides for controlling S. mosellana within an Integrated Pest Management program. A field study using insect pathogenic fungus Beauveria bassiana GHA, nematode Steinernema Jeltiae with Barricade polymer gel 1%, pyrethrin, combined formulations of B. bassiana GHA and pyrethrin, Jasmonic acid (JA) and chlorpyrifos (chemical check) was performed to determine to which extent they affect midge larval populations, kernel damage levels, grain yield, and quality, and the impacts on adult parasitoid Macroglenes penetrans populations. The results indicated that biopesticides JA and S. Jeltiae were the most effective in reducing larval populations and kernel damage levels, and produced a higher spring wheat yield when compared to the water control at both study locations (East Valier and North Valier, Montana, USA). Increased test weight in wheat had been recorded with two previous biopesticides at East Valier but not for North Valier, when compared over water control. These results were comparable in efficacy to the chlorpyrifos. This study also suggested that B. bassiana and pyrethrin may work synergistically, as exemplified by lower total larval populations and kernel damage levels when applied together. This study did not demonstrate the effect of any treatments on M. penetrans populations.     

The Promises and the Challenges of Biotransformations in Microflow

The challenges of transition toward the postpetroleum world shed light on the biocatalysis as the most sustainable way for the valorization of biobased raw materials. However, its industrial exploitation strongly relies on integration with innovative technologies such as microscale processing. Microflow devices remarkably accelerate biocatalyst screening and engineering, as well as evaluation of process parameters, and intensify biocatalytic processes in multiphase systems. The inherent feature of microfluidic devices to operate in a continuous mode brings additional interest for their use in chemoenzymatic cascade systems and in connection with the downstream processing units. Further steps toward automation and analytics integration, as well as computer‐assisted process development, will significantly affect the industrial implementation of biocatalysis and fulfill the promises of the bioeconomy. This review provides an overview of recent examples on implementation of microfluidic devices into various stages of biocatalytic process development comprising ultrahigh‐throughput biocatalyst screening, highly efficient biocatalytic process design including specific immobilization techniques for long‐term biocatalyst use, integration with other (bio)chemical steps, and/or downstream processing.