Biotechnological Perspective Research Articles

Expanding Pseudomonas taiwanensis VLB120's acyl-CoA portfolio: Propionate production in mineral salt medium.

As one of the main precursors, acetyl-CoA leads to the predominant production of even-chain products. From an industrial biotechnology perspective, extending the acyl-CoA portfolio of a cell factory is vital to producing industrial relevant odd-chain alcohols, acids, ketones and polyketides. The bioproduction of odd-chain molecules can be facilitated by incorporating propionyl-CoA into the metabolic network. The shortest pathway for propionyl-CoA production, which relies on succinyl-CoA catabolism encoded by the sleeping beauty mutase operon, was evaluated in Pseudomonas taiwanensis VLB120. A single genomic copy of the sleeping beauty mutase genes scpA, argK and scpB combined with the deletion of the methylcitrate synthase PVLB_08385 was sufficient to observe propionyl-CoA accumulation in this Pseudomonas. The chassis' capability for odd-chain product synthesis was assessed by expressing an acyl-CoA hydrolase, which enabled propionate synthesis. Three fed-batch strategies during bioreactor fermentations were benchmarked for propionate production, in which a maximal propionate titre of 2.8 g L-1 was achieved. Considering that the fermentations were carried out in mineral salt medium under aerobic conditions and that a single genome copy drove propionyl-CoA production, this result highlights the potential of Pseudomonas to produce propionyl-CoA derived, odd-chain products.

Inhibition of bacterial biofilms by the snake venom proteome

Snake venoms possess a range of pharmacological and toxicological activities. Here we evaluated the antibacterial and anti-biofilm activity against methicillin-susceptible and methicillin-resistant Staphylococcus aureus (MSSA and MRSA) of venoms from the Samar spitting cobra Naja samarensis and the Puff adder Bitis arietans. Both venoms prevented biofilm production by pathogenic S. aureus in a growth-independent manner, with the B. arietans venom being most potent. Fractionation showed the active molecule to be heat-labile and >10 kDa in size. Proteomic profiles of N. samarensis venom revealed neurotoxins and cytotoxins, as well as an abundance of serine proteases and three-finger toxins, while serine proteases, metalloproteinases and C-lectin types were abundant in B. arietans venom. These enzymes may have evolved to prevent bacteria colonising the snake venom gland. From a biomedical biotechnology perspective, they have valuable potential for anti-virulence therapy to fight antibiotic resistant microbes.

The human body from a biotechnological perspective in art contexts: active space of experimentation in constant transformation

Over the last three decades, the rise of new conceptualizations around the human body in connection with technology and machines has led to the development of new art practices in parallel with technological advances in the fields of biology, medicine and computing. This paper examines the human body in art contexts where biotechnology plays a relevant role in the generation of new conceptualizations around the human body topic. Thus, re-contextualized in social and cultural spheres and technical terms, the body will become redesigned, revamped, or augmented through the use of technological advances ranging from plastic surgery to robotic prosthetics. The human body is our vehicle of direct interaction with the environment. Its modification, alteration or expansion implies a redefinition of the existing limits between our corporality and the environment itself. Not only in the physical sense, but metaphysical and social. Hence, the introduced artists and strategies blur these limits and establish new relations through the body as a physical and social entity. In the search of new boundaries far beyond socio-cultural and natural limitations, they will reach new social, perceptual and conceptual statuses, far beyond aesthetic practices, focusing on the generation of political and social debates.

A Brief Atlas of Insulin.

Insulin is an essential factor for mammalian organisms: a regulator of glucose metabolism and other key signaling pathways. Insulin is also a multifunctional hormone whose absence can cause many diseases. Recombinant insulin is widely used in the treatment of diabetes. Understanding insulin, biosimilars, and biobetters from a holistic perspective will help pharmacologically user-friendly molecules design and develop personalized medicine-oriented therapeutic strategies for diabetes. Additionally, it helps to understand the underlying mechanism of other insulindependent metabolic disorders. The purpose of this atlas is to review insulin from a biotechnological, basic science, and clinical perspective, explain nearly all insulin-related disorders and their underlying molecular mechanisms, explore exogenous/recombinant production strategies of patented and research-level insulin/analogs, and highlight their mechanism of action from a structural perspective. Combined with computational analysis, comparisons of insulin and analogs also provide novel information about the structural dynamics of insulin.

Sensing seaweed settings: Making sense of a mixed-method design for sensory analysis

The focus of this paper is an interdisciplinary approach towards the consumption of novel seaweed foods. Combining social science, food science, and a chemical and biotechnological perspective, we ask the following questions: How do we gain knowledge about the consumer's approach to new marine products on the market? What kinds of methods are suitable and possible to use? In what way can we combine different kinds of methods and what can we learn from each other? In this paper we argue for a mixed methods approach, while ensuring flexibility and openness to disciplinary processes. Through a pilot study and by sharing empirical experiences, we explore the sensory properties of four different seaweed species, to investigate departure points for designing the larger interdisciplinary study about the consumption of novel seaweed products. Navigating the highly complex, interconnected production and consumption systems of seaweed, the question arises of how to reach a deeper understanding and more closely connect the research processes over the borders of different disciplines. We have taken our starting point in an approach that enabled a framework that encouraged engagement and learning throughout the research process.

In Vitro and In Vivo Cysticidal Effects of Carica Papaya Cell Suspensions.

Parasitic diseases are a major public health problem worldwide. Plant-derived products appear to be ideal candidates from a biotechnological perspective, being sustainable and environmentally friendly. The antiparasitic properties of Carica papaya have been attributed to some of its components, including papain and other compounds that are concentrated in the latex and seeds. This study demonstrated in vitro a high and insignificantly different cysticidal activity of soluble extract that was obtained after the disruption of nontransformed wild-type (WT) cells as well as transformed papaya calluses (PC-9, PC-12, and PC-23) and papaya cell suspensions (CS-9, CS-12, and CS-23). In vivo, cell suspensions of CS-WT and CS-23 that had been previously lyophilized were tested with respect to their cysticidal effects, compared with those of three commercial antiparasitic drugs. CS-WT and CS-23 together reduced the number of cysticerci, the number of buds, and the percentage of calcified cysticerci in a similar extent to albendazole and niclosamide, whereas ivermectin was less effective. Mice were then orally immunized with CS-23 that expressed the anti-cysticercal KETc7 antigen (10 μg/mouse), CS-WT (10 mg/mouse), or both together to evaluate their preventive properties. CS-23 and CS-WT significantly reduced the expected parasite and increased the percentage of calcified cysticerci as well as recovery, being more effective when employed together. The results reported in this study support the feasibility of the development of an anti-cysticercosis vaccine from cells of C. papaya in in vitro cultures, as they are a source of an anthelmintic, natural, and reproducible product.

Biotechnological perspective for wireless energy: H2-based power extraction from air

Despite its extreme scarcity, atmospheric H2 serves as an energy source for some prokaryotes. Recently, Grinter, Kropp, et al. reported the structural, biochemical, electrochemical, and spectroscopic elucidation of an underlying H2 catalyst, a [NiFe]-hydrogenase, which, owing to its extremely high affinity, facilitates the extraction of energy from ambient air.

Fungal Endophytes: A Storehouse of Bioactive Compounds.

Fungal endophytes are the microbial adaptations that usually enter the plant tissues during their life cycle without harming the host plants. They are found everywhere on earth and generally depend on the hosts by developing various symbiotic relationships, like mutualism, hostility, and parasitism on rare occasions, leading to the growth and rise in the nutrient content of the hosts. Endophytes can develop tolerance in host organisms against the stresses induced by either living or non-living agents. They may protect them from insects or pests by building resistance. Interestingly, endophytes can synthesize many phytohormones, natural medicinal compounds and several essential enzymes beneficial for biotechnological perspectives that can be obtained by culturing plant tissue in a suitable medium. These endophytes are a reservoir of many new active phytoconstituents, like alkaloids, phenolics, steroids, quinones, tannins, saponins, etc., which exhibit anticancer, antiinsecticidal, antioxidant, antibacterial, antiviral, antifungal, and many more properties. Exploring the new bioactive chemical entities from the endophytes may supply potent lead compounds for drug discovery to combat numerous disease conditions. Hence, the present review was carried out to explore the significance of the fungal endophytes and their medicinal, food, and cosmetic use.

Vitis vinifera genotyping toolbox to highlight diversity and germplasm identification.

The contribution of vine cultivation to human welfare as well as the stimulation of basic social and cultural features of civilization has been great. The wide temporal and regional distribution created a wide array of genetic variants that have been used as propagating material to promote cultivation. Information on the origin and relationships among cultivars is of great interest from a phylogenetics and biotechnology perspective. Fingerprinting and exploration of the complicated genetic background of varieties may contribute to future breeding programs. In this review, we present the most frequently used molecular markers, which have been used on Vitis germplasm. We discuss the scientific progress that led to the new strategies being implemented utilizing state-of-the-art next generation sequencing technologies. Additionally, we attempted to delimit the discussion on the algorithms used in phylogenetic analyses and differentiation of grape varieties. Lastly, the contribution of epigenetics is highlighted to tackle future roadmaps for breeding and exploitation of Vitis germplasm. The latter will remain in the top of the edge for future breeding and cultivation and the molecular tools presented herein, will serve as a reference point in the challenging years to come.

Post-Digital Learning for Rural Development: A Case Study of Open Biotec MOOCs in Colombia

This research presents an experience of post-digital learning of biotechnology in rural areas in Colombia by implementing a massive open online course (MOOC) for biotechnology education called Open Biotec. The objective was to improve Colombian rural development by creating learning communities around five topics: waste management, environmental biotechnology, bioprocesses, alternative energies, and bio-entrepreneurship. The study utilized various methods, including a socioeconomic diagnosis of a subregion of the department of Antioquia, Colombia, and the creation of MOOCs using the Action Research methodology. The pilot test of the training route in biotechnology involved the progressive development of the MOOC courses that comprise it. Around 918 students from rural areas were trained, and statistical analysis showed that the average grade of the students increased significantly from 6.13 to 7.53 and the median from 6.15 to 8.00. The study results demonstrate how the learning experience designed in the Open Biotec MOOC increases the degree of knowledge of students in this field of biotechnology, offering an opportunity to establish sustainable learning communities with participation and collaborative action mediated by virtual tools. The study concludes that rural education in Colombia could be strengthened with a training strategy for rural communities supported by MOOCs focused on the responsible use of local biodiversity from a biotechnological perspective.

Eurypsychrophilic acidophiles: From (meta)genomes to low-temperature biotechnologies.

Low temperature and acidic environments encompass natural milieus such as acid rock drainage in Antarctica and anthropogenic sites including drained sulfidic sediments in Scandinavia. The microorganisms inhabiting these environments include polyextremophiles that are both extreme acidophiles (defined as having an optimum growth pH < 3), and eurypsychrophiles that grow at low temperatures down to approximately 4°C but have an optimum temperature for growth above 15°C. Eurypsychrophilic acidophiles have important roles in natural biogeochemical cycling on earth and potentially on other planetary bodies and moons along with biotechnological applications in, for instance, low-temperature metal dissolution from metal sulfides. Five low-temperature acidophiles are characterized, namely, Acidithiobacillus ferriphilus, Acidithiobacillus ferrivorans, Acidithiobacillus ferrooxidans, "Ferrovum myxofaciens," and Alicyclobacillus disulfidooxidans, and their characteristics are reviewed. Our understanding of characterized and environmental eurypsychrophilic acidophiles has been accelerated by the application of "omics" techniques that have aided in revealing adaptations to low pH and temperature that can be synergistic, while other adaptations are potentially antagonistic. The lack of known acidophiles that exclusively grow below 15°C may be due to the antagonistic nature of adaptations in this polyextremophile. In conclusion, this review summarizes the knowledge of eurypsychrophilic acidophiles and places the information in evolutionary, environmental, biotechnological, and exobiology perspectives.

Time for a Change! A Spotlight on Many Neglected Facets of Sponge Microbial Biotechnology.

The sponge-microorganism partnership is one of the most successful symbiotic associations exploited from a biotechnological perspective. During the last thirty years, sponge-associated bacteria have been increasingly harnessed for bioactive molecules, notably antimicrobials and cytotoxic compounds. Unfortunately, there are gaps in sponge microbial biotechnology, with a multitude of applications being understudied or ignored. In this context, the current perspective aims to shed light on these underrated facets of sponge microbial biotechnology with a balance of existent reports and proposals for further research in the field. Our overview has showcased that the members of the sponge microbiome produce biomolecules whose usage can be valuable for several economically- relevant and demanding sectors. Outside the exhaustive search for antimicrobial secondary metabolites, sponge-associated microorganisms are gifted producers of antibiofilm, antivirulence and chronic diseases-attenuating substances highly envisaged by the pharmaceutical industry. Despite still at an infant stage of research, anti-ageing enzymes and pigments of special interest for the cosmetic and cosmeceutical sectors have also been reported from the sponge microbial symbionts. In a world urging for sustainability, sponge-associated microorganisms have been proven as fruitful resources for bioremediation, including recovery of heavy-metal contaminated areas, bioleaching processes, and as bioindicators of environmental pollution. In conclusion, we propose alternatives to better assess these neglected biotechnological applications of the sponge microbiome in the hope of sparking the interest of the scientific community toward their deserved exploitation.

Consulting with Māori during development of a point-of-care device; translational and experiential findings.

Aim: To consult with Māori on the design and development of a direct-to-consumer point-of-care (POC) device and gather views on point-of-care testing and biotechnology.&#x0D; Method: One-on-one interviews and small group hui with self-identified Māori university staff and students (n = 6) conducted by an early-career Māori scientist. &#x0D; Results: Key themes were the importance of achieving improved health outcomes for Māori through addressing known socioeconomic, geographic, and cultural factors that perpetuate health disparities. Other findings were the value of recognising the diversity in modern Māori identities, perspectives, and communities, as well as views on using synthetic biomolecules in medical devices and perceptions of biotechnology, and the potential for cultural over-engagement or misplaced focus in consultation. &#x0D; Conclusion: In this article, we describe our approach and experience of consultation led by a Māori lab-based scientist, and report unique perspectives of biotechnology from non-expert Māori academics for the first time. Direct-to-consumer POC testing may promote kaupapa Māori values such as tino rangatiratanga, whakawhanaungatanga and tikanga, which may help Māori overcome barriers to health care and testing, a key step in achieving improved health outcomes.

A Novel Strategy to Enhance Antioxidant Content in Saccharomyces Cerevisiae Based on Oxygen Pressure.

Antioxidant foods represent a potent lever to improve diets while creating value. Yet, their cultivation is often tied to a specific area and climate, limiting availability and increasing market cost. Therefore, microorganism-based antioxidant production emerges as a promising technology to solve these problems. In this view, a novel process was investigated for antioxidant accumulation in yeast culture. S. cerevisiae cells were exposed to various hyperbaric air conditions from 1 to 9 bar (A). Yeast cultures exhibited an increased reactive oxygen species content, which induced oxidative defense expression. After a few hours, reactive oxygen species levels decreased while antioxidant contents remained high, leading to a net increase in antioxidant power. At 6 bar (A), yeast achieved the highest net antioxidant power (phenolics content +48.3 ± 18.6 %, reducing power +120 ± 11.4 %) with an acceptable growth rate (0.27 h-1). Regarding time evolution, a 2 h exposure seems to be the optimum: cells have the lowest reactive oxygen species level while their antioxidant power is increased. From a biotechnological perspective, this finding highlights air pressure as an antioxidant-manipulating stress strategy. Moreover, the proposed process led to a patent that could potentially reduce energy and chemical consumption in such antioxidant accumulation processes.

Chlorella vulgaris acclimated cultivation under flashing light: An in-depth investigation under iso-actinic conditions

Chlorella vulgaris was cultivated under continuous and flashing light. Special care was taken in designing a culture device ensuring iso-actinic conditions, and a protocol ensuring acclimation. Continuous light cultures were carried out from 25 to 800 μmolPhotonPAR/m2/s. They served as control. Flashing light (square shape, duty cycle of 0.5) was applied with four frequencies: 0.1, 1, 10, and 100 Hz. The monitored outcomes were the culture growth rate, cell pigment content (chlorophylls, lutein, violaxanthin, and zeaxanthin), and photosynthetic apparatus (Fv/Fm and light curve). Acclimated cultures under continuous light showed photolimitation and photosaturation phases, without sign of photoinhibition, even under 800 μmolPhotonPAR/m2/s. Frequencies of 1 Hz and above induced little to no difference between cells acclimated under flashing light and their control (same amount of continuous light). Indeed, cells exhibited the same growth rate, similar absolute pigment composition, and light curve characteristics. They only differ by a lower chlorophyll b to chlorophyll a ratio at high intensity, suggesting an acclimation strategy favoring an increased number of photosystems instead of an increased light-harvesting capability. Conversely, cells cultivated at 0.1 Hz showed a lower growth rate and an incapacity to adapt efficiently to high incident illumination. From a biotechnological perspective, these results support the idea that high frequency flashing light, in the tested conditions, does not bring benefits with respect to the same average amount of continuous light. Consequently, it may ease the design of large scale photobioreactors, where light/dark cycles are inevitable.

Environmental and Biotechnological Perspectives on Hydrolases

Micro-organisms are able to survive in high salt concentrations because they have developed diverse biochemical, structural and physiological modifications, allowing the catalytic synthesis of proteins with interesting physicochemical and structural properties. When opposed to chemical catalysts, enzymes are protein-based catalysts that offer significant advantages. Enzymes are viewed as a greener alternative to conventional chemical catalysis in industrial processes since they are biodegradable, reusable, and do not produce excessive waste products. Enzymes are useful biological instruments for bioremediation that are both environmentally acceptable and bio based. An essential class of enzymes called hydrolases encourages the depolymerisation of macromolecules, which is crucial for the metabolism of nutrients. They have properties that make them desirable for biotechnological applications, in particular for environment protection, where hydrolaseproducing organisms can be used in the removal of contaminants through a process known as bioremediation. They are highly effective catalysts, frequently secreted into the surrounding medium, and the majority do not require cofactors. Hydrolases present an opportunity to investigate the conformational variation that underlies the wide range of biological functions that these enzymes perform because they are the most numerous and diverse class of enzymes. The best choice for eliminating pollutants from the environment may be microbial hydrolases. All facets of life contain these enzymes, which are crucial for key biological activities like the breakdown of cellulosic biomass, viral pathogenesis, antibacterial defence, and typical cellular functioning. The fact that certain lysosomal storage illnesses have been linked to deficits in these enzymes, as well as the potential industrial uses of highly effective glycoside hydrolases. The isolation of active enzymes against particular pollutants comes from microorganisms that have been exposed to contaminated locations and those contaminants specifically. For the bioremediation of contaminants, several enzymes extracted from various species have been utilised. A viable approach to finding more effective and affordable instruments for the clean-up of contaminants would be to identify novel enzymes and new subtypes with specified physicochemical properties. Keywords: Enzymes; Hydrolases; Catalyst; Microbes; Applicati

Antioxidant and Antimicrobial Properties of Selected Red Seaweeds from Central Portugal

Throughout the ages, macroalgae have provided humankind with elements beneficial to human health, and often with bioactive abilities. Yet, while today we fully acknowledge such potential, especially that of the most widely known species, an even greater number of species remain unaknowledged. This holds particularly true for the highly diverse phylum Rhodophyta (red seaweeds) and, therefore, the present study aims to unveil the antioxidant and the antimicrobial potential of twelve red seaweed species collected in central Portugal. Results obtained from the antioxidant assays ABTS and TPC highlighted the high scavenging capacity of the coralline algae Corallina officinalis, Ellisolandia elongata and Amphiroa rigida, and the high phenolic content of Porphyra umbilicalis, whereas the antimicrobial analyses through MIC determination emphasized the activities of Sphaerococcus coronopifolius and Mesophyllum lichenoides against, respectively, Bacillus subtilis and Saccharomyces cerevisiae. This study raised awareness of the bioactive potential waiting to be discovered regarding less known Rhodophyta species, such as Amphiroa rigida and Mesophyllum lichenoides. Therefore, we believe this study provides extra steps in pinpointing Rhodophyta species with bioactive potential, encouraging further studies tailored toward a biotechnological perspective, and, ultimately, influencing current perspectives regarding the exploration of seaweeds.

Exploring Potentials for Bioresource and Bioenergy Recovery from Vinasse, the “New” Protagonist in Brazilian Sugarcane Biorefineries

Vinasse management in biorefineries bears a burden for sugarcane industries. Despite its high potassium-related fertilizer potential, a series of negative environmental impacts is expected to occur in long-term soil applications of vinasse through fertirrigation. Conversely, a high biodegradable organic content characterizes vinasse as a potential substrate for bioresource and bioenergy recovery from numerous (bio)technological perspectives. This review presents the alternative approaches proposed for sugarcane vinasse management in Brazil, with special attention dedicated to the role of anaerobic digestion as the core conversion step. The suitability of applying phase separation, i.e., the separation of fermentation from methanogenesis in sequential reactors, is discussed in detail. Laboratory and full-scale experiences were considered to discuss the energetic potential of sugarcane vinasse through biogas generation. With a national installed capacity of up to 1603 MW, energy from vinasse could replace half of the coal-derived electricity in Brazil. Meanwhile, investing in vinasse fermentation to obtain soluble organic metabolites could provide more than 10 g L−1 of (iso)butyrate. This is the first review addressing the potential use of sugarcane vinasse in anaerobic biorefineries that discusses applications far beyond conventional biogas production, and encourages the rational use of vinasse as a raw material for bioprocesses, either in short- or long-term scenarios.

Bacillales: From Taxonomy to Biotechnological and Industrial Perspectives.

For a long time, the genus Bacillus has been known and considered among the most applicable genera in several fields. Recent taxonomical developments resulted in the identification of more species in Bacillus-related genera, particularly in the order Bacillales (earlier heterotypic synonym: Caryophanales), with potential application for biotechnological and industrial purposes such as biofuels, bioactive agents, biopolymers, and enzymes. Therefore, a thorough understanding of the taxonomy, growth requirements and physiology, genomics, and metabolic pathways in the highly diverse bacterial order, Bacillales, will facilitate a more robust designing and sustainable production of strain lines relevant to a circular economy. This paper is focused principally on less-known genera and their potential in the order Bacillales for promising applications in the industry and addresses the taxonomical complexities of this order. Moreover, it emphasizes the biotechnological usage of some engineered strains of the order Bacillales. The elucidation of novel taxa, their metabolic pathways, and growth conditions would make it possible to drive industrial processes toward an upgraded functionality based on the microbial nature.

Functional genomics of the lactic acid bacterium Limosilactobacillus fermentum LAB-1: metabolic, probiotic and biotechnological perspectives

A genome-based systematic analysis was conducted to characterize the metabolic, probiotic, fitness, and safety properties of Limosilactobacillus fermentum LAB-1, a lactic acid bacterium demonstrating strong antimicrobial effects against clinical pathogens. Gene functional characterization revealed a large number of genes for carbohydrate metabolism and a heterofermentative system for carbon dissimilation. Genes for intact pyruvate oxidation, pentose phosphate, and PRPP biosynthetic pathways were identified. Substantial carbohydrate-active enzymes and transporters were also predicted. Metabolic reconstruction revealed complete sets of enzymes for arginine, lysine, methionine, threonine, proline, and ornithine biosynthesis. The bacterium harbors a diverse range of peptidases, and a large variety of peptide and amino acid uptake systems. It encodes restriction-modification and CRISPR-Cas systems for protection against phage infections and carries a wide spectrum of stress proteins for adaptation in the gut and industrial conditions. Genes related to the biosynthesis of B-group and K vitamins were identified allowing its application for novel bio-enriched food production. Other beneficial traits of probiotic and industrial importance such as production of flavor compounds, exopolysaccharide, acetoin, and butanediol were identified. Three antimicrobial peptides were predicted which showed >98% sequence-identity to experimentally validated bacteriocins. Negative traits such as transmissible antibiotic resistance, pathogenicity or virulence appeared to be absent suggesting the strain to be considered safe. The genome analysis will allow precisely targeted laboratory research and full exploitation of the probiotic potentials towards functional-food, biotechnology and health-related applications.