Biotechnological Capabilities Research Articles

Exploring the biotechnological potential of Acinetobacter soli ANG344B: A novel bacterium for 2-phenylethanol production

A bacterium, Acinetobacter soli ANG344B, isolated from river water, exhibited an exceptional capacity to produce 2-phenylethanol (2-PE) using L-phenylalanine (L-Phe) as a precursor—a capability typically observed in yeasts rather than bacteria. Bioreactor experiments were conducted to evaluate the production performance, using glucose as the carbon source for cellular growth and L-Phe as the precursor for 2-PE production. Remarkably, A. soli ANG344B achieved a 2-PE concentration of 2.35 ± 0.26 g/L in just 24.5 h of cultivation, exhibiting a global volumetric productivity of 0.10 ± 0.01 g/L.h and a production yield of 0.51 ± 0.01 g2-PE/gL-Phe, a result hitherto reported only for yeasts. These findings position A. soli ANG344B as a highly promising microorganism for 2-PE production.Whole-genome sequencing of A. soli strain ANG344 revealed a genome size of 3.52 Mb with a GC content of 42.7 %. Utilizing the Rapid Annotation using Subsystem Technology (RAST) server, 3418 coding genes were predicted, including genes coding for enzymes previously associated with the metabolic pathway of 2-PE production in other microorganisms, yet unreported in Acinetobacter species. Through gene mapping, 299 subsystems were identified, exhibiting 30 % subsystem coverage. The whole genome sequence data was submitted to NCBI GeneBank with the BioProject ID PRJNA982713. These draft genome data offer significant potential for exploiting the biotechnological capabilities of A. soli strain ANG344 and for conducting further comparative genomic studies.

Genomic insights of Fictibacillus terranigra sp. nov., a versatile metabolic bacterium from Amazonian Dark Earths.

The Amazon rainforest, a hotspot for biodiversity, is a crucial research area for scientists seeking novel microorganisms with ecological and biotechnological significance. A key region within the Amazon rainforest is the Amazonian Dark Earths (ADE), noted for supporting diverse plant and microbial communities, and its potential as a blueprint for sustainable agriculture. This study delineates the isolation, morphological traits, carbon source utilization, and genomic features of Fictibacillus terranigra CENA-BCM004, a candidate novel species of the Fictibacillus genus isolated from ADE. The genome of Fictibacillus terranigra was sequenced, resulting in 16 assembled contigs, a total length of 4,967,627bp, and a GC content of 43.65%. Genome annotation uncovered 3315 predicted genes, encompassing a wide range of genes linked to various metabolic pathways. Phylogenetic analysis indicated that CENA-BCM004 is a putative new species, closely affiliated with other unidentified Fictibacillus species and Bacillus sp. WQ 8-8. Moreover, this strain showcased a multifaceted metabolic profile, revealing its potential for diverse biotechnological applications. It exhibited capabilities to antagonize pathogens, metabolize multiple sugars, mineralize organic matter compounds, and solubilize several minerals. These insights substantially augment our comprehension of microbial diversity in ADE and underscore the potential of Fictibacillus terranigra as a precious resource for biotechnological endeavors. The genomic data generated from this study will serve as a foundational resource for subsequent research and exploration of the biotechnological capabilities of this newly identified species.

Genome sequences of seven bacterial isolates collected from soil as part of a microbiology lab course.

Soil is a source for diverse microbes that possess useful biotechnological capabilities. Here, we report the genome sequences of seven bacterial isolates from the species Exiguobacterium acetylicum, Rossellomorea marisflavi, Delftia acidovorans, Pseudomonas aeruginosa, Bacillus sp., and Bacillus toyonensis (two isolates) cultured from Dallas/Fort Worth metroplex soil samples.

Экологические особенности и биотехнологические возможности почвенных актинобактерий (обзор)

Actinobacteria are gram-positive aerobic and facultative anaerobic bacteria that tend to form branching hyphae capable of developing into mycelium. Their main role is in the decomposition of complex polymers in the soil: lignin, cellulose, chitin, humus substances; fertility formation; production of antibiotic substances; accumulation of biologically active compounds. Actinobacteria secrete into the environment a complex of secondary exometabolites of various compositions with aliphatic, carbocyclic, heterocyclic, nitrogenous, oxygen-containing and sulfur-containing compounds. Most of the isolated secondary metabolites have antibiotic, antimicrobial, antiviral, herbicidal, insecticidal properties. Biotechnological capabilities of soil actinobacteria producing biologically active substances with a wide range of environmental effects are used in the development of multifunctional biologics for various industries. Actinobacteria play a crucial role in optimizing the growing conditions of plants, and therefore there is a natural interest in studying the factors that influence the relationship between actinobacteria and various crops in specific soil conditions. The development of this issue is an important link in the ecological characterization of the soil microbial complex and it will help to develop the theoretical foundations for the targeted regulation of the interactions of actinobacteria with phytopathogenic microorganisms and insect pests. The purpose of this work is to analyze the ecological features and biotechnological capabilities of soil actinobacteria. The review describes the distribution of actinobacteria in various types of soils, including those with the presence of pollutant; the composition of the metabolites produced; relationships with other microorganisms, as well as biotechnological properties and the possibility of using actinobacteria for the development of biological products for agroecosystems.

Review on Microalgae Potential Innovative Biotechnological Applications

Novel compounds can be found in marine creatures, many of which have amazing biotechnological capabilities. Microalgae, in particular, have a drawn interest as a potential basis for new industrial creation routes. Many biologically active compounds, such as antioxidants, immunostimulants, antivirals, antibiotics, hem agglutinates, polyunsaturated fatty acids, peptides, proteins, biofuels, and pigments, are derived from these species. Recently, there has been a rise in interest in microalgal biotechnology to create beneficial, sustainable, and ecologically friendly bioproducts. Microalgae biomass is in high demand for a wide range of potential uses, most of which are now the subject of ongoing research. Microalgae are important groups of photosynthetic organisms that use light and carbon dioxide more efficiently than terrestrial plants to produce biomass and use it for biotechnological purposes such as environmental protection, biofuel production, pharmaceutical production, human food supplements, animal feed components, coronavirus treatments, and so on. This paper presents an overview of current advancements in the application of microalgal biotechnology in several industries.

TECHNOLOGICAL CAPABILITIES IN MEXICAN AGROBIOTECHNOLOGY: A PROPOSAL FOR THEIR MEASUREMENT

This article proposes a taxonomy to measure technological capabilities in biotechnology, specifically in agrobiotechnology. Based on this taxonomy, the case of Mexican companies is analyzed to identify the type and level of technological capabilities through a survey conducted between 2018 and 2019. The results show significant differences by type of company, it stands out that startups and multinational companies report predominantly basic levels in the areas of decision-making, linking, research and development, and generation of processes and products. While small and medium-sized companies have predominantly intermediate levels. These differences point to the need to generate differentiated industrial and technological policies that address the diversity of agrobiotechnology companies.

Thorsmoerkia curvula gen. et spec. nov. (Trebouxiophyceae, Chlorophyta), a semi-terrestrial microalga from Iceland exhibits high levels of unsaturated fatty acids

A terrestrial green alga was isolated at Iceland, and the strain (SAG 2627) was described for its morphology and phylogenetic position and tested for biotechnological capabilities. Cells had a distinctly curved, crescent shape with conical poles and a single parietal chloroplast. Phylogenetic analyses of 18S rDNA and rbcL markers placed the strain into the Trebouxiophyceae (Chlorophyta). The alga turned out to belong to an independent lineage without an obvious sister group within the Trebouxiophyceae. Based on morphological and phylogenetic data, the strain was described as a new genus and species, Thorsmoerkia curvula gen. et sp. nov. Biomass was generated in column reactors and subsequently screened for promising metabolites. Growth was optimized by pH-regulated, episodic CO2 supplement during the logarithmic growth-phase, and half of the biomass was thereafter exposed to nitrogen and phosphate depletion. The biomass yield reached up to 53.5 mg L−1 day−1. Fatty acid (FA) production peaked at 24 mg L−1 day−1 and up to 83% of all FAs were unsaturated. At the end of the log phase, approximately 45% of dry mass were lipids, including eicosapentaenoic acid. Carotenoid production reached up to 2.94 mg L−1 day−1 but it was halted during the stress phase. The N-linked glycans of glycoproteins were assessed to reveal chemotaxonomic patterns. The study demonstrated that new microalgae can be found at Iceland, potentially suitable for applied purposes. The advantage of T. curvula is its robustness and that significant amounts of lipids are already accumulated during log phase, making a subsequent stress exposure dispensable.

Isolation and characterization of a novel rhamnolipid producer Pseudomonas sp. LGMS7 from a highly contaminated site in Ain El Arbaa region of Ain Temouchent, Algeria.

The online version contains supplementary material available at 10.1007/s13205-021-02751-6.

Recombinant cellulase of Caulobacter crescentus: potential applications for biofuels and textile industries

Caulobacter crescentus (C. crescentus) has a single celA gene encoding the bacterial cellulase (EC 3.2.1.4) of the GH9 family that is induced at transcriptional levels (q-PCR) by agroindustrial residues. Corn straw (1%, w/v) was more efficient in inducing bacterial mRNA-celA expression than corn cob (1%, w/v) and a parallel protein expression profile was observed, since enzyme activity was also higher in presence of corn straw in the bacterium. In order to evaluate the biotechnological role of cellulase from C. crescentus (NA1000), the celA gene of 1,878 bp (CCNA_02310) was cloned in the pTrCHisA vector and successfully expressed in Escherichia coli. The recombinant cellulase CelA purified showed 73 kDa, specificity to CMC, optimal pH of 5.5 and pH stability of 6, optimal temperature at 40 °C and thermostability at 35 °C, KM = 0.66 mg/mL and VMax = 2.41 U/mg/min. CelA was strongly activated by ions Mn2+ (70%), Sn2+ (49%) and Co2+ (46%) and was efficient in the hydrolysis of corn straw and corn cob (1%; w/v) releasing 2.62 µmol/mL and 1.02 µmol/mL of reducing sugar, respectively, being an alternative for the production of chemicals and biofuels. CelA was applied directly to the jeans fiber and scanning electron micrographs (SEM) generated showed a clean and smooth surface of the fibers compared to the control, indicating the application of enzyme for the biopolishing of denim jeans. This work supports a set of biotechnological capabilities of cellulase from C. crescentus suggested by the bacterial genomic sequence that has not been described yet.

The use of antifungal oat‐sourdough lactic acid bacteria to improve safety and technological functionalities of the supplemented wheat bread

AbstractIn the present study, predominant lactic acid bacteria (LAB) were isolated from oat sourdough. Then, the isolates were screened based on their in vitro antifungal activity. Subsequently, biotechnological capabilities of the selected LAB were evaluated in wheat bread supplemented with controlled fermented oat containing the isolate. Pediococcus pentosaceus was molecular identified as predominant antifungal isolate. Based on our results, fermented oat not only significantly (p < .05) reduced the mold expansion on the produced breads, but also improved the quality attributes of the product. Crumb porosity and antioxidant capacity of the supplemented bread revealed the positive effects of the isolate on textural and functional characteristics of the enriched bread. Wheat bread supplemented with controlled fermented oat had also the highest 2,2‐diphenyl‐1‐picrylhydrazyl radical scavenging activity compared to the other samples. Accordingly, the potential applications of P. pentosaceus isolate as profunctional starter culture in processing of mixed wheat‐oat sourdough bread were verified.

Twentieth-century nonproliferation meets twenty-first-century biotechnology

ABSTRACT This article explores emerging science and technology advances relevant to the 1972 Biological and Toxin Weapons Convention (BWC), and existing and needed frameworks for their identification, risk assessment, and evaluation of benefit. Threats from biological weapons continue to be a major concern as state and non-state actors have developed, used, or expressed interest in these types of weapons. International nonproliferation instruments and related efforts in health security, specifically the 2005 International Health Regulations and the 2014 Global Health Security Agenda, recognize deliberate biological incidents as one of three threats to address (the others being natural and accidental biological events). To date, these instruments and their subsequent regional and national-level implementation efforts focus primarily on pathogens and toxins as biological threat agents. Unlike the other instruments, the BWC focuses on preventing the diversion of peaceful and prophylactic uses of biology to the development, production, stockpiling, or dissemination and delivery of biological weapons. Accordingly, the BWC recognizes the importance of scientific and technological advances in enabling different actors to develop or disseminate biological agents, altering the risk profile of deliberate biological threats. To identify and discuss advances that may affect implementation of the BWC, the US National Academy of Sciences and the BWC Implementation Support Unit conducted or sponsored several activities to explore science and technologies that may be most relevant to the BWC. However, the biotechnology landscape continues to change drastically, expanding the focus of security risks beyond pathogens and toxins to include other biological data and materials, such as synthetic organisms. Factors promoting the development of biotechnology capabilities include new funders and funding models, practitioners from other disciplines leveraging the tools of biology, new nations investing in the biological sciences, and research leveraging advances in engineering, computer, data, materials, physical, and chemical sciences. These advances may reveal new capabilities that significantly alter biological nonproliferation efforts, including both new security threats and benefits to society.

USING 3D PRINTING TECHNOLOGY TO FULL-SCALE SIMULATION OF THE UPPER RESPIRATORY TRACT

The project "Implementation of rapid prototyping for modelling the upper respiratory tract in normal and typical pathologies" investigates the urgent problem of improving the reliability of diagnosis and effectiveness of treatment of disorders of the nasal breathing. Possibilities of modern 3D-printing technology for creation of individual natural anatomical models of the upper respiratory tract and determination of their aerodynamic characteristics are considered. The characteristics of the laminar boundary layer of the air flow in the parietal region of the nasal cavity are investigated under different modes of breathing in normal and with typical disorders of the nasal breathing. The concept of investigation of the aerodynamic indices of the anatomical structures of the respiratory system by the results of test tests of individual full-scale 3D models, obtained by the data of spiral computed tomography, is being developed. Theoretical bases of the method of computer planning of restorative rhinosurgical interventions in patients with chronic diseases of the nasal cavity are grounded, based on the change of the configuration of the anatomical structures of the nasal cavity taking into account the aerodynamic parameters of respiration. Modern distance learning and testing tools are being created to demonstrate the technology developed, to provide theoretical knowledge, practical skills and to solve situational tasks for a wide range of specialists. Development and research of natural patterns of the upper respiratory tract allows for supplementing and expanding the knowledge about the aerodynamic characteristics of the nasal cavity, to make decisions about therapy in a short period of time. Experience of the Laboratory of the Institute for Multiphase Processes (IMP) of the Leibniz Universität Hannover (LUH) in the development and use of rapid prototyping capabilities in biotechnology will provide technical support to the project.

A Broad-Host-Range CRISPRi Toolkit for Silencing Gene Expression in Burkholderia.

Genetic tools are critical to dissecting the mechanisms governing cellular processes, from fundamental physiology to pathogenesis. Members of the genus Burkholderia have potential for biotechnological applications but can also cause disease in humans with a debilitated immune system. The lack of suitable genetic tools to edit Burkholderia GC-rich genomes has hampered the exploration of useful capacities and the understanding of pathogenic features. To address this, we have developed CRISPR interference (CRISPRi) technology for gene silencing in Burkholderia, testing it in B.cenocepacia, B.multivorans, and B.thailandensis. Tunable expression was provided by placing a codon-optimized dcas9 from Streptococcus pyogenes under control of a rhamnose-inducible promoter. As a proof of concept, the paaABCDE operon controlling genes necessary for phenylacetic acid degradation was targeted by plasmid-borne gRNAs, resulting in near complete inhibition of growth on phenylacetic acid as the sole carbon source. This was supported by reductions in paaA mRNA expression. The utility of CRISPRi to probe other functions at the single cell level was demonstrated by knocking down phbC and fliF, which dramatically reduces polyhydroxybutyrate granule accumulation and motility, respectively. As a hallmark of the mini-CTX system is the broad host-range of integration, we putatively identified 67 genera of Proteobacteria that might be amenable to modification with our CRISPRi toolkit. Our CRISPRi toolkit provides a simple and rapid way to silence gene expression to produce an observable phenotype. Linking genes to functions with CRISPRi will facilitate genome editing with the goal of enhancing biotechnological capabilities while reducing Burkholderia's pathogenic arsenal.

Azospirillum brasilense Increases CO2 Fixation on Microalgae Scenedesmus obliquus, Chlorella vulgaris, and Chlamydomonas reinhardtii Cultured on High CO2 Concentrations.

Mutualism interactions of microalgae with other microorganisms are widely used in several biotechnological processes since symbiotic interaction improves biotechnological capabilities of the microorganisms involved. The interaction of the bacterium Azospirillum brasilense was assessed with three microalgae genus, Scenedesmus, Chlorella, and Chlamydomonas, during CO2 fixation under high CO2 concentrations. The results in this study have demonstrated that A. brasilense maintained a mutualistic interaction with the three microalgae assessed, supported by the metabolic exchange of indole-3-acetic acid (IAA) and tryptophan (Trp), respectively. Besides, CO2 fixation increased, as well as growth and cell compound accumulation, mainly carbohydrates, in each microalgae evaluated, interacting with the bacterium. Overall, these results propose the mutualism interaction of A. brasilense with microalgae for improving biotechnological processes based on microalgae as CO2 capture and their bio-refinery capacity.

Genome Mutational and Transcriptional Hotspots Are Traps for Duplicated Genes and Sources of Adaptations.

Gene duplication generates new genetic material, which has been shown to lead to major innovations in unicellular and multicellular organisms. A whole-genome duplication occurred in the ancestor of Saccharomyces yeast species but 92% of duplicates returned to single-copy genes shortly after duplication. The persisting duplicated genes in Saccharomyces led to the origin of major metabolic innovations, which have been the source of the unique biotechnological capabilities in the Baker’s yeast Saccharomyces cerevisiae. What factors have determined the fate of duplicated genes remains unknown. Here, we report the first demonstration that the local genome mutation and transcription rates determine the fate of duplicates. We show, for the first time, a preferential location of duplicated genes in the mutational and transcriptional hotspots of S. cerevisiae genome. The mechanism of duplication matters, with whole-genome duplicates exhibiting different preservation trends compared to small-scale duplicates. Genome mutational and transcriptional hotspots are rich in duplicates with large repetitive promoter elements. Saccharomyces cerevisiae shows more tolerance to deleterious mutations in duplicates with repetitive promoter elements, which in turn exhibit higher transcriptional plasticity against environmental perturbations. Our data demonstrate that the genome traps duplicates through the accelerated regulatory and functional divergence of their gene copies providing a source of novel adaptations in yeast.

Preservation of protein expression systems at elevated temperatures for portable therapeutic production.

Many biotechnology capabilities are limited by stringent storage needs of reagents, largely prohibiting use outside of specialized laboratories. Focusing on a large class of protein-based biotechnology applications, we address this issue by developing a method for preserving cell-free protein expression systems for months above room temperature. Our approach realizes unprecedented long-term stability at elevated temperatures by leveraging the sugar alcohol trehalose, a simple, low-cost, open-air drying step, and strategic separation of reaction components during drying. The resulting preservation capacity enables efficient production of a wide range of on-demand proteins under adverse conditions, for instance during emergency outbreaks or in remote locations. To demonstrate application potential, we use cell-free reagents subjected to months of exposure at 37°C and atmospheric conditions to produce sufficient concentrations of a pyocin protein to kill Pseudomonas aeruginosa, a troublesome pathogen for traumatic and burn wound injuries. Our work makes possible new biotechnology applications that demand ruggedness and scalability.

Microbial community structure of two freshwater sponges using Illumina MiSeq sequencing revealed high microbial diversity.

Sponges are primitive metazoans that are known to harbour diverse and abundant microbes. All over the world attempts are being made to exploit these microbes for their biotechnological potential to produce, bioactive compounds and antimicrobial peptides. However, the majority of the studies are focussed on the marine sponges and studies on the freshwater sponges have been neglected so far. To increase our understanding of the microbial community structure of freshwater sponges, microbiota of two fresh water sponges namely, Eunapius carteri and Corvospongilla lapidosa is explored for the first time using Next Generation Sequencing (NGS) technology. Overall the microbial composition of these sponges comprises of 14 phyla and on an average, more than 2900 OTUs were obtained from C. lapidosa while E. carteri showed 980 OTUs which is higher than OTUs obtained in the marine sponges. Thus, our study showed that, fresh water sponges also posses highly diverse microbial community than previously thought and it is distinct from the marine sponge microbiota. The present study also revealed that microbial community structure of both the sponges is significantly different from each other and their respective water samples. In the present study, we have detected many bacterial lineages belonging to Firmicutes, Actinobacteria, Proteobacteria, Planctomycetes, etc. that are known to produce compounds of biotechnological importance. Overall, this study gives insight into the microbial composition of the freshwater sponges which is highly diverse and needs to be studied further to exploit their biotechnological capabilities.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0211-2) contains supplementary material, which is available to authorized users.

CellF: a neuron-driven music synthesiser for real-time performance

ABSTRACTcellF is the world’s first neuron-driven synthesiser. It is a collaborative project at the cutting-edge of experimental art and music that brings together artists, musicians, designers and scientists to create a cybernetic self-portrait. cellF is an autonomous, bio-analogue electronic musical instrument designed to operate independently and interact with human musicians. The instrument is controlled by a bio-engineered neural network or ‘brain’ derived from skin cells using induced pluripotent stem cell (iPSC) technology that is housed in a custom-built synthesiser ‘body’. This paper introduces the cellF project highlighting the background, bio-technology, embodiment, design and performance capabilities.

The criticality of high-resolution N-linked carbohydrate assays and detailed characterization of antibody effector function in the context of biosimilar development

Accurate measurement and functional characterization of antibody Fc domain N-linked glycans is critical to successful biosimilar development. Here, we describe the application of methods to accurately quantify and characterize the N-linked glycans of 2 IgG1 biosimilars with effector function activity, and show the potential pitfalls of using assays with insufficient resolution. Accurate glycan assessment was combined with glycan enrichment using lectin chromatography or production with glycosylation inhibitors to produce enriched pools of key glycan species for subsequent assessment in cell-based antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity effector function assays. This work highlights the challenges of developing high-quality biosimilar candidates and the need for modern biotechnology capabilities. These results show that high-quality analytics, combined with sensitive cell-based assays to study in vivo mechanisms of action, is an essential part of biosimilar development.

The emergence of the nanobiotechnology industry

The confluence of nanotechnology and biotechnology provides significant commercial opportunities. By identifying, classifying and tracking firms with capabilities in both biotechnology and nanotechnology over time, we analyse the emergence and evolution of the global nanobiotechnology industry.