Optimization of de novo transcriptome assembly from high-throughput short read sequencing data improves functional annotation for non-model organisms

Background: The k-mer hash length is a key factor affecting the output of de novo transcriptome assembly packages using de Bruijn graph algorithms. Assemblies constructed with varying single k-mer choices might result in the loss of unique contiguous sequences (contigs) and relevant biological information. A common solution to this problem is the clustering of single k-mer assemblies. Even though annotation is one of the primary goals of a transcriptome assembly, the success of assembly strategies does not consider the impact of k-mer selection on the annotation output. This study provides an in-depth k-mer selection analysis that is focused on the degree of functional annotation achieved for a non-model organism where no reference genome information is available. Individual k-mers and clustered assemblies (CA) were considered using three representative software packages. Pair-wise comparison analyses (between individual k-mers and CAs) were produced to reveal missing Kyoto Encyclopedia of Genes and Genomes (KEGG) ortholog identifiers (KOIs), and to determine a strategy that maximizes the recovery of biological information in a de novo transcriptome assembly. Results: Analyses of single k-mer assemblies resulted in the generation of various quantities of contigs and functional annotations within the selection window of k-mers (k-19 to k-63). For each k-mer in this window, generated assemblies contained certain unique contigs and KOIs that were not present in the other k-mer assemblies. Producing a non-redundant CA of k-mers 19 to 63 resulted in a more complete functional annotation than any single k-mer assembly. However, a fraction of unique annotations remained (~0.19 to 0.27% of total KOIs) in the assemblies of individual k-mers (k-19 to k-63) that were not present in the non-redundant CA. A workflow to recover these unique annotations is presented. Conclusions: This study demonstrated that different k-mer choices result in various quantities of unique contigs per single k-mer assembly which affects biological information that is retrievable from the transcriptome. This undesirable effect can be minimized, but not eliminated, with clustering of multi-k assemblies with redundancy removal. The complete extraction of biological information in de novo transcriptomics studies requires both the production of a CA and efforts to identify unique contigs that are present in individual k-mer assemblies but not in the CA

A review of nature-based solutions for resource recovery in cities

Our modern cities are resource sinks designed on the current linear economic model which recovers very little of the original input. As the current model is not sustainable, a viable solution is to recover and reuse parts of the input. In this context, resource recovery using nature-based solutions (NBS) is gaining popularity worldwide. In this specific review, we focus on NBS as technologies that bring nature into cities and those that are derived from nature, using (micro)organisms as principal agents, provided they enable resource recovery. The findings presented in this work are based on an extensive literature review, as well as on original results of recent innovation projects across Europe. The case studies were collected by participants of the COST Action Circular City, which includes a portfolio of more than 92 projects. The present review article focuses on urban wastewater, industrial wastewater, municipal solid waste and gaseous effluents, the recoverable products (e.g., nutrients, nanoparticles, energy), as well as the implications of source-separation and circularity by design. The analysis also includes assessment of the maturity of different technologies (technology readiness level) and the barriers that need to be overcome to accelerate the transition to resilient, self-sustainable cities of the future

Transcriptomic analysis of the oleaginous microalga Neochloris oleoabundans reveals metabolic insights into triacylglyceride accumulation

Background: The lack of sequenced genomes for oleaginous microalgae limits our understanding of the mechanisms these organisms utilize to become enriched in triglycerides. Here we report the de novo transcriptome assembly and quantitative gene expression analysis of the oleaginous microalga Neochloris oleoabundans, with a focus on the complex interaction of pathways associated with the production of the triacylglycerol (TAG) biofuel precursor. Results: After growth under nitrogen replete and nitrogen limiting conditions, we quantified the cellular content of major biomolecules including total lipids, triacylglycerides, starch, protein, and chlorophyll. Transcribed genes were sequenced, the transcriptome was assembled de novo, and the expression of major functional categories, relevant pathways, and important genes was quantified through the mapping of reads to the transcriptome. Over 87 million, 77 base pair high quality reads were produced on the Illumina HiSeq sequencing platform. Metabolite measurements supported by genes and pathway expression results indicated that under the nitrogen-limiting condition, carbon is partitioned toward triglyceride production, which increased fivefold over the nitrogen-replete control. In addition to the observed overexpression of the fatty acid synthesis pathway, TAG production during nitrogen limitation was bolstered by repression of the β-oxidation pathway, up-regulation of genes encoding for the pyruvate dehydrogenase complex which funnels acetyl-CoA to lipid biosynthesis, activation of the pentose phosphate pathway to supply reducing equivalents to inorganic nitrogen assimilation and fatty acid biosynthesis, and the up-regulation of lipases—presumably to reconstruct cell membranes in order to supply additional fatty acids for TAG biosynthesis. Conclusions: Our quantitative transcriptome study reveals a broad overview of how nitrogen stress results in excess TAG production in N. oleoabundans, and provides a variety of genetic engineering targets and strategies for focused efforts to improve the production rate and cellular content of biofuel precursors in oleaginous microalgae.Connecticut Center for Advanced Technology (Fuel Diversification Grant, by the National Science Foundation Grant #0854322)Yale Climate and Energy InstituteYale Institute for Biospheric StudiesYale University. Biomedical High Performance Computing CenterNational Institutes of Health (U.S.) (NIH Grant # RR19895

Microfluidics for microalgal biotechnology

Ozdalgic, Berin/0000-0003-0113-541X; Kiraz, Alper/0000-0001-7977-1286; Ustun, Merve/0000-0002-3883-4065; Tasoglu, Savas/0000-0003-4604-217X; Haznedaroglu, Berat Zeki/0000-0002-0081-8801; Rahmani Dabbagh, Sajjad/0000-0001-8888-6106Microalgae have expanded their roles as renewable and sustainable feedstocks for biofuel, smart nutrition, biopharmaceutical, cosmeceutical, biosensing, and space technologies. They accumulate valuable biochemical compounds from protein, carbohydrate, and lipid groups, including pigments and carotenoids. Microalgal biomass, which can be adopted for multivalorization under biorefinery settings, allows not only the production of various biofuels but also other value-added biotechnological products. However, state-of-the-art technologies are required to optimize yield, quality, and the economical aspects of both upstream and downstream processes. As such, the need to use microfluidic-based devices for both fundamental research and industrial applications of microalgae, arises due to their microscale sizes and dilute cultures. Microfluidics-based devices are superior to their competitors through their ability to perform multiple functions such as sorting and analyzing small amounts of samples (nanoliter to picoliter) with higher sensitivities. Here, we review emerging applications of microfluidic technologies on microalgal processes in cell sorting, cultivation, harvesting, and applications in biofuels, biosensing, drug delivery, and nutrition.Alexander von Humboldt-StiftungAlexander von Humboldt Foundation; Marie Sklodowska-Curie Individual Fellowship award [101003361]; Royal Academy Newton-Katip Celebi Transforming Systems Through Partnership Award [120N019]; TUBITAK 2232 International Fellowship for Outstanding Researchers AwardTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [118C391]Alexander von Humboldt-Stiftung, Grant/Award Number: Research Fellowship for Experienced Researchers; Marie Sklodowska-Curie Individual Fellowship award, Grant/Award Number: 101003361; Royal Academy Newton-Katip Celebi Transforming Systems Through Partnership Award, Grant/Award Number: 120N019; TUBITAK 2232 International Fellowship for Outstanding Researchers Award, Grant/Award Number: 118C39

Carbon nanotube bundling: influence on layer-by-layer assembly and antimicrobial activity

Peer reviewe

Advanced Methods for Natural Products Discovery: Bioactivity Screening, Dereplication, Metabolomics Profiling, Genomic Sequencing, Databases and Informatic Tools, and Structure Elucidation

Natural Products (NP) are essential for the discovery of novel drugs and products for numerous biotechnological applications. The NP discovery process is expensive and time-consuming, having as major hurdles dereplication (early identification of known compounds) and structure elucidation, particularly the determination of the absolute configuration of metabolites with stereogenic centers. This review comprehensively focuses on recent technological and instrumental advances, highlighting the development of methods that alleviate these obstacles, paving the way for accelerating NP discovery towards biotechnological applications. Herein, we emphasize the most innovative high-throughput tools and methods for advancing bioactivity screening, NP chemical analysis, dereplication, metabolite profiling, metabolomics, genome sequencing and/or genomics approaches, databases, bioinformatics, chemoinformatics, and three-dimensional NP structure elucidation

Nature-Based Units as Building Blocks for Resource Recovery Systems in Cities

Cities are producers of high quantities of secondary liquid and solid streams that are still poorly utilized within urban systems. In order to tackle this issue, there has been an ever-growing push for more efficient resource management and waste prevention in urban areas, following the concept of a circular economy. This review paper provides a characterization of urban solid and liquid resource flows (including water, nutrients, metals, potential energy, and organics), which pass through selected nature-based solutions (NBS) and supporting units (SU), expanding on that characterization through the study of existing cases. In particular, this paper presents the currently implemented NBS units for resource recovery, the applicable solid and liquid urban waste streams and the SU dedicated to increasing the quality and minimizing hazards of specific streams at the source level (e.g., concentrated fertilizers, disinfected recovered products). The recovery efficiency of systems, where NBS and SU are combined, operated at a micro- or meso-scale and applied at technology readiness levels higher than 5, is reviewed. The importance of collection and transport infrastructure, treatment and recovery technology, and (urban) agricultural or urban green reuse on the quantity and quality of input and output materials are discussed, also regarding the current main circularity and application challenges

Valorization of Marine Waste: Use of Industrial By-Products and Beach Wrack Towards the Production of High Added-Value Products

Biomass is defined as organic matter from living organisms represented in all kingdoms. It is recognized to be an excellent source of proteins, polysaccharides and lipids and, as such, embodies a tailored feedstock for new products and processes to apply in green industries. The industrial processes focused on the valorization of terrestrial biomass are well established, but marine sources still represent an untapped resource. Oceans and seas occupy over 70% of the Earth’s surface and are used intensively in worldwide economies through the fishery industry, as logistical routes, for mining ores and exploitation of fossil fuels, among others. All these activities produce waste. The other source of unused biomass derives from the beach wrack or washed-ashore organic material, especially in highly eutrophicated marine ecosystems. The development of high-added-value products from these side streams has been given priority in recent years due to the detection of a broad range of biopolymers, multiple nutrients and functional compounds that could find applications for human consumption or use in livestock/pet food, pharmaceutical and other industries. This review comprises a broad thematic approach in marine waste valorization, addressing the main achievements in marine biotechnology for advancing the circular economy, ranging from bioremediation applications for pollution treatment to energy and valorization for biomedical applications. It also includes a broad overview of the valorization of side streams in three selected case study areas: Norway, Scotland, and the Baltic Sea.</p&gt

Valorization of Marine Waste : Use of Industrial By-Products and Beach Wrack Towards the Production of High Added-Value Products

Biomass is defined as organic matter from living organisms represented in all kingdoms. It is recognized to be an excellent source of proteins, polysaccharides and lipids and, as such, embodies a tailored feedstock for new products and processes to apply in green industries. The industrial processes focused on the valorization of terrestrial biomass are well established, but marine sources still represent an untapped resource. Oceans and seas occupy over 70% of the Earth's surface and are used intensively in worldwide economies through the fishery industry, as logistical routes, for mining ores and exploitation of fossil fuels, among others. All these activities produce waste. The other source of unused biomass derives from the beach wrack or washed-ashore organic material, especially in highly eutrophicated marine ecosystems. The development of high-added-value products from these side streams has been given priority in recent years due to the detection of a broad range of biopolymers, multiple nutrients and functional compounds that could find applications for human consumption or use in livestock/pet food, pharmaceutical and other industries. This review comprises a broad thematic approach in marine waste valorization, addressing the main achievements in marine biotechnology for advancing the circular economy, ranging from bioremediation applications for pollution treatment to energy and valorization for biomedical applications. It also includes a broad overview of the valorization of side streams in three selected case study areas: Norway, Scotland, and the Baltic Sea.</p&gt