The Impact of Non-Concentrated Storage on the Centrifugation Yield of Microchloropsis gaditana: A Pilot-Scale Study.

Non-concentrated algae storage can bridge the period between algae harvesting and processing while avoiding the stress conditions associated with the concentration step required for concentrate storage. This study aimed to examine organic matter losses during the non-concentrated storage of Microchloropsis gaditana at pilot-scale. Algae cultures (400-500 L) were stored for up to 12 days either at an 8 °C target temperature or at 19 °C as the average temperature. The centrifugation yield of stored algal cultures decreased from day 5 or day 8 onwards for all storage conditions. After 12 days, the centrifugation yields were between 57% and 93% of the initial yields. Large differences in centrifugation yields were noted between the algae batches. The batch-to-batch difference outweighed the effect of storage temperature, and the highest yield loss was observed for the 8 °C cooled algae batch. The analysis of stored algae before and after centrifugation suggested that the decreasing yields were not related to respiration losses, but rather, the decreasing efficiency with which organic matter is collected during the centrifugation step.

The nutritional profile, mineral content and heavy metal uptake of yellow mealworm reared with supplementation of agricultural sidestreams.

Insect farming, a potential approach to deal with the increasing global protein demand, is a new activity in the Western world with many unanswered questions regarding product quality and safety. Insects may fulfill an important role in a circular economy by upcycling biowaste into valuable biomass. About half of the total mass of mealworm feeding substrates exists out of wet feed. This can be sourced from biowaste, increasing the sustainability of insect farming. This paper reports on the nutritional profile of yellow mealworm, Tenebrio molitor, reared with supplementation of organic sidestreams. These included unsold vegetables, potato cuttings, fermented chicory roots and horticultural foliage. The evaluation was performed by analyzing proximate compositions, fatty acid profiles, mineral and heavy metal contents. Mealworms fed with potato cuttings doubled their fat content and increased saturated and mono-unsaturated fatty acids. Providing fermented chicory roots increased the mineral content and accumulated heavy metals. Additionally, the uptake of minerals by mealworms was selective as only calcium, iron and manganese concentrations increased. Adding vegetable mix or horticultural foliage to the diet did not significantly change the nutritional profile. In conclusion, sidestreams were successfully recycled into protein-rich biomass and their nutrient content and bio-availability influenced the composition of mealworms.

Pilot-Scale Cultivation of the Snow Alga Chloromonas typhlos in a Photobioreactor.

The most studied and cultivated microalgae have a temperature optimum between 20 and 35°C. This temperature range hampers sustainable microalgae growth in countries with colder periods. To overcome this problem, psychrotolerant microalgae, such as the snow alga Chloromonas typhlos, can be cultivated during these colder periods. However, most of the research work has been carried out in the laboratory. The step between laboratory-scale and large-scale cultivation is difficult, making pilot-scale tests crucial to gather more information. Here, we presented a successful pilot-scale growth test of C. typhlos. Seven batch mode growth periods were compared during two longer growth tests in a photobioreactor of 350 L. We demonstrated the potential of this alga to be cultivated at colder ambient temperatures. The tests were performed during winter and springtime to compare ambient temperature and sunlight influences. The growth and CO2 usage were continuously monitored to calculate the productivity and CO2 fixation efficiency. A maximum dry weight of 1.082 g L-1 was achieved while a maximum growth rate and maximum daily volumetric and areal productivities of 0.105 d-1, 0.110 g L-1 d-1, and 2.746 g m-2 d-1, respectively, were measured. Future tests to optimize the cultivation of C. typhlos and production of astaxanthin, for example, will be crucial to explore the potential of biomass production of C. typhlos on a commercial scale.

Protein-Protein Interactions in Candida albicans.

Despite being one of the most important human fungal pathogens, Candida albicans has not been studied extensively at the level of protein-protein interactions (PPIs) and data on PPIs are not readily available in online databases. In January 2018, the database called "Biological General Repository for Interaction Datasets (BioGRID)" that contains the most PPIs for C. albicans, only documented 188 physical or direct PPIs (release 3.4.156) while several more can be found in the literature. Other databases such as the String database, the Molecular INTeraction Database (MINT), and the Database for Interacting Proteins (DIP) database contain even fewer interactions or do not even include C. albicans as a searchable term. Because of the non-canonical codon usage of C. albicans where CUG is translated as serine rather than leucine, it is often problematic to use the yeast two-hybrid system in Saccharomyces cerevisiae to study C. albicans PPIs. However, studying PPIs is crucial to gain a thorough understanding of the function of proteins, biological processes and pathways. PPIs can also be potential drug targets. To aid in creating PPI networks and updating the BioGRID, we performed an exhaustive literature search in order to provide, in an accessible format, a more extensive list of known PPIs in C. albicans.

A High-Throughput Candida albicans Two-Hybrid System.

Candida albicans is a human fungal pathogen that does not follow the universal codon usage, as it translates the CUG codon into serine rather than leucine. This makes it difficult to study protein-protein interactions using the standard yeast two-hybrid (Y2H) system in the model organism Saccharomyces cerevisiae Due to the lack of adapted tools, only a small number of protein-protein interactions (PPIs) have been detected or studied using C. albicans-optimized tools despite the importance of PPIs to understand cell biology. However, with the sequencing of the whole genome of C. albicans, the availability of an ORFeome collection containing 5,099 open reading frames (ORFs) in Gateway-adapted donor vectors, and the creation of a Gateway-compatible C. albicans-specific two-hybrid (C2H) system, it became possible to study protein-protein interactions on a larger scale using C. albicans itself as the model organism. Erroneous translations are hereby eliminated compared to using the S. cerevisiae Y2H system. Here, we describe the technical adaptations and the first application of the C2H system for a high-throughput screen, thus making it possible to screen thousands of PPIs at once in C. albicans itself. This first, small-scale high-throughput screen, using Pho85 as a bait protein against 1,646 random prey proteins, yielded one interacting partner (Pcl5). The interaction found with the high-throughput setup was further confirmed with a low-throughput C2H experiment and with a coimmunoprecipitation (co-IP) experiment.IMPORTANCECandida albicans is a major fungal pathogen, and due to the rise of fungal infections and emerging resistance to the limited antifungals available, it is important to develop novel and more specific antifungals. Protein-protein interactions (PPIs) can be applied as very specific drug targets. However, because of the aberrant codon usage of C. albicans, the traditional yeast two-hybrid system in Saccharomyces cerevisiae is difficult to use, and only a limited number of PPIs have been described in C. albicans To overcome this, a C. albicans two-hybrid (C2H) system was developed in 2010. The current work describes, for the first time, the application of the C2H system in a high-throughput setup. We hereby show the usefulness of the C2H system to investigate and detect PPIs in C. albicans, making it possible to further elucidate protein networks in C. albicans, which has the potential to lead to the development of novel antifungals which specifically disrupt PPIs important for virulence.

Generating genomic platforms to study Candida albicans pathogenesis.

The advent of the genomic era has made elucidating gene function on a large scale a pressing challenge. ORFeome collections, whereby almost all ORFs of a given species are cloned and can be subsequently leveraged in multiple functional genomic approaches, represent valuable resources toward this endeavor. Here we provide novel, genome-scale tools for the study of Candida albicans, a commensal yeast that is also responsible for frequent superficial and disseminated infections in humans. We have generated an ORFeome collection composed of 5099 ORFs cloned in a Gateway™ donor vector, representing 83% of the currently annotated coding sequences of C. albicans. Sequencing data of the cloned ORFs are available in the CandidaOrfDB database at http://candidaorfeome.eu. We also engineered 49 expression vectors with a choice of promoters, tags and selection markers and demonstrated their applicability to the study of target ORFs transferred from the C. albicans ORFeome. In addition, the use of the ORFeome in the detection of protein-protein interaction was demonstrated. Mating-compatible strains as well as Gateway™-compatible two-hybrid vectors were engineered, validated and used in a proof of concept experiment. These unique and valuable resources should greatly facilitate future functional studies in C. albicans and the elucidation of mechanisms that underlie its pathogenicity.