Biotechnological Strategies Research Articles

Synthetic organic antibiotics residues as emerging contaminants waste-to-resources processing for a circular economy in China: Challenges and perspective

Synthetic antibiotics have been known for years to combat bacterial antibiotics. But their overuse and resistance have become a concern recently. The antibiotics reach the environment, including soil from the manufacturing process and undigested excretion by cattle and humans. It leads to overburden and contamination of the environment. These organic antibiotics remain in the environment for a very long period. During this period, antibiotics come in contact with various flora and fauna. The ill manufacturing practices and inadequate wastewater treatment cause a severe problem to the water bodies. After pretreatment from pharmaceutical industries, the effluents are released to the water bodies such as rivers. Even after pretreatment, effluents contain a significant number of antibiotic residues, which affect the living organisms living in the water bodies. Ultimately, river contaminated water reaches the ocean, spreading the contamination to a vast environment. This review paper discusses the impact of synthetic organic contamination on the environment and its hazardous effect on health. In addition, it analyzes and suggests the biotechnological strategies to tackle organic antibiotic residue proliferation. Moreover, the degradation of organic antibiotic residues by biocatalyst and biochar is analyzed. The circular economy approach for waste-to-resource technology for organic antibiotic residue in China is analyzed for a sustainable solution. Overall, the significant challenges related to synthetic antibiotic residues and future aspects are analyzed in this review paper.

Alginate encapsulated nanobio-hybrid system enables improvement of photocatalytic biohydrogen production in the presence of oxygen

Bacterial encapsulation for the generation of fuels and other valuable chemicals through fermentation emerges as a novel biotechnological strategy that provides significant advantages over the use of free cells. In the present study, we established an optimal alginate encapsulated TiO2/Escherichia coli hybrid system by varying the initial concentrations of alginate (2–5%) and initial inoculum concentration (OD600 0.5–2.5) to form the hydrogel beads for efficient production of hydrogen in the presence of oxygen with the given visible-light intensity of 2000 W m−2. The maximum hydrogen production of 2.8 mmol H2/mmol glucose was obtained with hydrogel beads consisting of 4% sodium alginate and initial inoculum with a cell density of 1.5 OD600. Natural sunlight could also be used to produce hydrogen under the same optimized condition. The optimization of the encapsulated nano bio-hybrid system with respect to alginate and inoculum concentration allows for the enhancement of hydrogen production even in the presence of oxygen.

Utilization of Soybean Oil Waste for a High-Level Production of Ceramide by a Novel Phospholipase C as an Environmentally Friendly Process

Ceramide is a natural functional ingredient as food additive and medicine that has attracted extensive attention in the food, medical, and cosmetic industries. Here, we developed a biotechnological strategy based on a recombinant whole-cell biocatalyst for efficiently producing ceramide from crude soybean oil sediment (CSOS) waste. A novel phospholipase C (PLCac) from Acinetobacter calcoaceticus isolated from soil samples was identified and characterized. Furthermore, recombinant Komagataella phaffii displaying PLCac (dPLCac) on the cell surface was constructed as a whole-cell biocatalyst with better thermostability (30-60 °C) and pH stability (8.0-10.0) to successfully produce ceramide. After synergistical optimization of reaction time and dPLCac dose, the ceramide yield of hydrolyzing from CSOS using dPLCac was 51% (the theoretical maximum yield of converting sphingomyelin, ∼70%) and the relative yield was over 50% after seven consecutive 4 h batches under the optimized conditions. Our study provides a potentially promising strategy for the commercial production of ceramide.

Arbuscular mycorrhizal fungi species improve the fatty acids profile and nutrients status of soybean cultivars grown under drought stress.

To investigate the effects of arbuscular mycorrhizal fungi (AMF) species on the absorption and distribution of mineral nutrients in soybean cultivars under drought stress, an experiment was carried out through a factorial method in the form of randomized complete blocks with six replicates in 2020. Experimental factors include: drought stress at three irrigation levels (well-watered [WW], medium stress [MS] and severe stress [SS], i.e., 100%, 70% and 40% FC), soybean cultivars at two levels (Sepideh and Williams), and mycorrhizae application at four levels (nonapplication, Funneliformis mosseae, Glomus hoi, Rhizophagus intraradices). The results indicated that drought stress increased the concentration of grain potassium (K) and sodium (Na), leaf K, stem and leaf Na, and decreased the concentrations of grain phosphorus (P), copper (Cu), and zinc (Zn), leaf P, stem and leaf Cu, and manganese (Mn). In addition, there was no significant difference in terms of concentrations of grain K. The highest reduction was observed under SS conditions (40% FC). G. hoi colonization increased the concentrations of grain P and Zn, leaf K and Cu and stem Mn under SS conditions (40% FC). In addition, compared to Sepideh cv., Williams cv. showed higher stress resistance. Seed oil content decreased in the plants exposed to drought stress. Severe drought treatments have a deleterious effect on seed fatty acid composition, resulting in enhanced linoleic, oleic and linolenic acids. AMF colonization is a useful tool for improving the plant nutrient uptake, fatty acid profile, efficiency of resource utilization and stabilizing yield, hence reducing the production risks of crops grown under drought stress conditions. It was concluded that AMF colonization should be employed to help alleviate the adverse effects of drought stress. AMF colonization is an effective biotechnological strategy that can alter nutrient uptake and fatty acid composition and enhance oil quality in soybean cultivars under drought conditions.

Natural 5-Aminolevulinic Acid: Sources, Biosynthesis, Detection and Applications.

5-Aminolevulinic acid (5-ALA) is the key precursor for the biosynthesis of tetrapyrrole compounds, with wide applications in medicine, agriculture and other burgeoning fields. Because of its potential applications and disadvantages of chemical synthesis, alternative biotechnological methods have drawn increasing attention. In this review, the recent progress in biosynthetic pathways and regulatory mechanisms of 5-ALA synthesis in biological hosts are summarized. The research progress on 5-ALA biosynthesis via the C4/C5 pathway in microbial cells is emphasized, and the corresponding biotechnological design strategies are highlighted and discussed in detail. In addition, the detection methods and applications of 5-ALA are also reviewed. Finally, perspectives on potential strategies for improving the biosynthesis of 5-ALA and understanding the related mechanisms to further promote its industrial application are conceived and proposed.

Carotenoid fortification of zucchini fruits using a viral RNA vector.

Carotenoids are health-promoting metabolites in livestock and human diets. Some important crops have been genetically modified to increase their content. Although the usefulness of transgenic plants to alleviate nutritional deficiencies is obvious, their social acceptance has been controversial. Here, we demonstrate an alternative biotechnological strategy for carotenoid fortification of edible fruits in which no transgenic DNA is involved. A viral RNA vector derived from zucchini yellow mosaic virus (ZYMV) was modified to express a bacterial phytoene synthase (crtB), and inoculated to zucchini (Cucurbita pepo L.) leaves nurturing pollinated flowers. After the viral vector moved to the developing fruit and expressed crtB, the rind and flesh of the fruits developed yellow-orange rather than green color. Metabolite analyses showed a substantial enrichment in health-promoting carotenoids, such as α- and β-carotene (provitamin A), lutein and phytoene, in both rind and flesh. Although this strategy is perhaps not free from controversy due to the use of genetically modified viral RNA, our work does demonstrate the possibility of metabolically fortifying edible fruits using an approach in which no transgenes are involved.

Classical and Next-Generation Vaccine Platforms to SARS-CoV-2: Biotechnological Strategies and Genomic Variants.

Several coronaviruses (CoVs) have been identified as human pathogens, including the α-CoVs strains HCoV-229E and HCoV-NL63 and the β-CoVs strains HCoV-HKU1 and HCoV-OC43. SARS-CoV, MERS-CoV, and SARS-CoV-2 are also classified as β-coronavirus. New SARS-CoV-2 spike genomic variants are responsible for human-to-human and interspecies transmissibility, consequences of adaptations of strains from animals to humans. The receptor-binding domain (RBD) of SARS-CoV-2 binds to receptor ACE2 in humans and animal species with high affinity, suggesting there have been adaptive genomic variants. New genomic variants including the incorporation, replacement, or deletion of the amino acids at a variety of positions in the S protein have been documented and are associated with the emergence of new strains adapted to different hosts. Interactions between mutated residues and RBD have been demonstrated by structural modelling of variants including D614G, B.1.1.7, B1.351, P.1, P2; other genomic variants allow escape from antibodies generated by vaccines. Epidemiological and molecular tools are being used for real-time tracking of pathogen evolution and particularly new SARS-CoV-2 variants. COVID-19 vaccines obtained from classical and next-generation vaccine production platforms have entered clinicals trials. Biotechnology strategies of the first generation (attenuated and inactivated virus–CoronaVac, CoVaxin; BBIBP-CorV), second generation (replicating-incompetent vector vaccines–ChAdOx-1; Ad5-nCoV; Sputnik V; JNJ-78436735 vaccine-replicating-competent vector, protein subunits, virus-like particles–NVX-CoV2373 vaccine), and third generation (nucleic-acid vaccines–INO-4800 (DNA); mRNA-1273 and BNT 162b (RNA vaccines) have been used. Additionally, dendritic cells (LV-SMENP-DC) and artificial antigen-presenting (aAPC) cells modified with lentiviral vector have also been developed to inhibit viral activity. Recombinant vaccines against COVID-19 are continuously being applied, and new clinical trials have been tested by interchangeability studies of viral vaccines developed by classical and next-generation platforms.

Understanding the Modus Operandi of Class II KNOX Transcription Factors in Secondary Cell Wall Biosynthesis.

Lignocellulosic biomass from the secondary cell walls of plants has a veritable potential to provide some of the most appropriate raw materials for producing second-generation biofuels. Therefore, we must first understand how plants synthesize these complex secondary cell walls that consist of cellulose, hemicellulose, and lignin in order to deconstruct them later on into simple sugars to produce bioethanol via fermentation. Knotted-like homeobox (KNOX) genes encode homeodomain-containing transcription factors (TFs) that modulate various important developmental processes in plants. While Class I KNOX TF genes are mainly expressed in the shoot apical meristems of both monocot and eudicot plants and are involved in meristem maintenance and/or formation, Class II KNOX TF genes exhibit diverse expression patterns and their precise functions have mostly remained unknown, until recently. The expression patterns of Class II KNOX TF genes in Arabidopsis, namely KNAT3, KNAT4, KNAT5, and KNAT7, suggest that TFs encoded by at least some of these genes, such as KNAT7 and KNAT3, may play a significant role in secondary cell wall formation. Specifically, the expression of the KNAT7 gene is regulated by upstream TFs, such as SND1 and MYB46, while KNAT7 interacts with other cell wall proteins, such as KNAT3, MYB75, OFPs, and BLHs, to regulate secondary cell wall formation. Moreover, KNAT7 directly regulates the expression of some xylan synthesis genes. In this review, we summarize the current mechanistic understanding of the roles of Class II KNOX TFs in secondary cell wall formation. Recent success with the genetic manipulation of Class II KNOX TFs suggests that this may be one of the biotechnological strategies to improve plant feedstocks for bioethanol production.

Opportunities and challenges for integrating the development of sustainable polymer materials within an international circular (bio)economy concept

HighlightsThe production and consumption of commodity polymers have been an indispensable part of the development of our modern society. Owing to their adjustable properties and variety of functions, polymer-based materials will continue playing important roles in achieving the Sustainable Development Goals (SDG)s, defined by the United Nations, in key areas such as healthcare, transport, food preservation, construction, electronics, and water management. Considering the serious environmental crisis, generated by increasing consumption of plastics, leading-edge polymers need to incorporate two types of functions: Those that directly arise from the demands of the application (e.g. selective gas and liquid permeation, actuation or charge transport) and those that enable minimization of environmental harm, e.g., through prolongation of the functional lifetime, minimization of material usage, or through predictable disintegration into non-toxic fragments. Here, we give examples of how the incorporation of a thoughtful combination of properties/functions can enhance the sustainability of plastics ranging from material design to waste management. We focus on tools to measure and reduce the negative impacts of plastics on the environment throughout their life cycle, the use of renewable sources for their synthesis, the design of biodegradable and/or recyclable materials, and the use of biotechnological strategies for enzymatic recycling of plastics that fits into a circular bioeconomy. Finally, we discuss future applications for sustainable plastics with the aim to achieve the SDGs through international cooperation.Leading-edge polymer-based materials for consumer and advanced applications are necessary to achieve sustainable development at a global scale. It is essential to understand how sustainability can be incorporated in these materials via green chemistry, the integration of bio-based building blocks from biorefineries, circular bioeconomy strategies, and combined smart and functional capabilities.Graphic abstract

Antioxidant Potential and Enhancement of Bioactive Metabolite Production in In Vitro Cultures of Scutellaria lateriflora L. by Biotechnological Methods.

Studies carried out using three different in vitro assays and a biological setting (Escherichia coil) demonstrated the antioxidant activity of Scutellaria lateriflora microshoot extract. Moreover, the extract exhibited no toxicity in a brine shrimp lethality bioassay. These results indicated that microshoots are a rich, safe source of antioxidants, which encouraged us to enhance their production in vitro. In agar and agitated cultures, two biotechnological strategies were applied: feeding the cultures with the biogenetic precursors of the phenolics—phenylalanine and tyrosine, and eliciting them with methyl jasmonate. Specific Scutellaria flavonoids and verbascoside were analysed by HPLC. Feeding with precursors (1 g/L) in agar cultures decreased the production of the metabolites. In agitated cultures, different concentrations of precursors (1.0–2.5 g/L) and the elicitor (10; 50; 100 µM) were tested. Additionally, parallel feeding with the precursor and elicitor in a concentration of 50 µM were applied. The best strategy for total flavonoid and verbascoside production was phenylalanine feeding (1.5 g/L), max. 3765 and 475 mg/100 g DW, respectively, after 7 days. This is the first report documenting the high antioxidant production in S. lateriflora microshoots after feeding with phenylalanine. Moreover, for the first time, bioreactor cultures were successfully maintained, obtaining attractive results (max. total flavonoid content 2348 and verbascoside 485 mg/100 g DW).

A fungal protease named AsES triggers antiviral immune responses and effectively restricts virus infection in arabidopsis and Nicotiana benthamiana plants.

Plants have evolved complex mechanisms to fight against pathogens. Among these mechanisms, pattern-triggered immunity (PTI) relies on the recognition of conserved microbe- or pathogen-associated molecular patterns (MAMPs or PAMPs, respectively) by membrane-bound receptors. Indeed, PTI restricts virus infection in plants and, in addition, BRI1-associated kinase 1 (BAK1), a central regulator of PTI, plays a role in antiviral resistance. However, the compounds that trigger antiviral defences, along with their molecular mechanisms of action, remain mostly elusive. Herein, we explore the role of a fungal extracellular subtilase named AsES in its capacity to trigger antiviral responses. In this study, we obtained AsES by recombinant expression, and evaluated and characterized its capacity to trigger antiviral responses against Tobacco mosaic virus (TMV) by performing time course experiments, analysing gene expression, virus movement and callose deposition. The results of this study provide direct evidence that exogenous treatment with recombinant AsES increases a state of resistance against TMV infection, in both arabidopsis and Nicotiana benthamiana plants. Also, the antiviral PTI response exhibited by AsES in arabidopsis is mediated by the BAK1/SERK3 and BKK1/SERK4 co-receptors. Moreover, AsES requires a fully active salicylic acid (SA) signalling pathway to restrict the TMV movement by inducing callose deposition. Additionally, treatment with PSP1, a biostimulant based on AsES as the active compound, showed an increased resistance against TMV in N. benthamiana and tobacco plants. AsES is a fungal serine protease which triggers antiviral responses relying on a conserved mechanism by means of the SA signalling pathway and could be exploited as an effective and sustainable biotechnology strategy for viral disease management in plants.

RICE BRAN; A POTENTIAL COMMODITY

Rice crop is cultivated in large amount worldwide and the outward layer of rice grain is bran that is the most important rice by-product. Rice bran and its oil carry bioactive components in large amount and these compounds are used to treat number of diseases and health defects, it is also used on industrial scale like in cosmetics. Bioactive components like γ-Oryzanol, flavonoids, phytic acid, phenolics, phytic acid are present in rice bran. Rice wheat has incredible potential in the food business, particularly in the assembling of useful nourishments, such as active baking products. The fortified rice or its components are used as additives in number of food matrices such as pizza, cookies, bread, beverages, tuna oil, powdered beef and milk for practical and nutritious purposes. Rice bran can be used to develop low fat and high fibrous products of bakery. It is also used in meat emulsion and industrial batter mixtures. In incorporation to the physical wellbeing that are provided by rice bran. Rice bran used as a high fiber diet can provide texture, firmness, emulsifying properties, gelling and certain stabilizing properties. Nourishments covered with a strong layer of rice regularly retain low fat during browning however the modest quantity of fat discovered normally in rice wheat fiber also go about as a flavor transporter. Boiled rice contains a lot of amino acids that are important for humans and because of it is used as a new nutritional constituent. Rice bran contains about ten- fifteen percent increased protein. The protein composition construct on rice bran can help control the disorders associated with a protein diet. In addition, unique anticancer and hypoallergenic properties make rice bran protein higher than cereal protein. Owing to its noteworthy nutritive and helpful esteem, rice bran may upgrade well‐being, as well as decrease the hazard of malady, giving wellbeing benefits and progressing quality of life. Hence, rice bran can be considered a super nourishment and utilitarian nourishment. In any case, the genuine potential of utilitarian bran can be created utilizing unused biotechnological strategy.

Approaches and determinants to sustainably improve crop production

AbstractPlant scientists and farmers are facing major challenges in providing food and nutritional security for a growing population, while preserving natural resources and biodiversity. Moreover, this should be done while adapting agriculture to climate change and by reducing its carbon footprint. To address these challenges, there is an urgent need to breed crops that are more resilient to suboptimal environments. Huge progress has recently been made in understanding the physiological, genetic and molecular bases of plant nutrition and environmental responses, paving the way towards a more sustainable agriculture. In this review, we present an overview of these progresses and strategies that could be developed to increase plant nutrient use efficiency and tolerance to abiotic stresses. As illustrated by many examples, they already led to promising achievements and crop improvements. Here, we focus on nitrogen and phosphate uptake and use efficiency and on adaptation to drought, salinity and heat stress. These examples first show the necessity of deepening our physiological and molecular understanding of plant environmental responses. In particular, more attention should be paid to investigate stress combinations and stress recovery and acclimation that have been largely neglected to date. It will be necessary to extend these approaches from model plants to crops, to unravel the relevant molecular targets of biotechnological or genetic strategies directly in these species. Similarly, sustained efforts should be done for further exploring the genetic resources available in these species, as well as in wild species adapted to unfavourable environments. Finally, technological developments will be required to breed crops that are more resilient and efficient. This especially relates to the development of multiscale phenotyping under field conditions and a wide range of environments, and use of modelling and big data management to handle the huge amount of information provided by the new molecular, genetic and phenotyping techniques.

Cholesterol Lowering Biotechnological Strategies: From Monoclonal Antibodies to Antisense Therapies. A Pre-Clinical Perspective Review.

In recent years, the increase in available genetic information and a better understanding of the genetic bases of dyslipidemias has led to the identification of potential new avenues for therapies. Additionally, the development of new technologies has presented the key for developing novel therapeutic strategies targeting not only proteins (e.g., the monoclonal antibodies and vaccines) but also the transcripts (from antisense oligonucleotides (ASOs) to small interfering RNAs) or the genomic sequence (gene therapies). These pharmacological advances have led to successful therapeutic improvements, particularly in the cardiovascular arena because we are now able to treat rare, genetically driven, and previously untreatable conditions (e.g, familial hypertriglyceridemia or hyperchylomicronemia). In this review, the pre-clinical pharmacological development of the major biotechnological cholesterol lowering advances were discussed, describing facts, gaps, potential future steps forward, and therapeutic opportunities.

Exploring an enhanced rhizospheric phenomenon for pluricontaminated soil remediation: Insights from tripartite metatranscriptome analyses

Phytoremediation involving the use of microorganisms with tolerant plant species represents a new frontier for on-site remediation of pluricontaminated soils. In this study, the effectiveness of a biotechnological strategy, involving the use of Festuca arundinacea and a pool of microorganisms, was assessed by a mesocosm experiment and an in-depth rhizospheric metatranscriptomic analysis. The chemical profile of mesocosm soil at the end of the experiment (240 days) showed that the decrease of trace elements such as Cd, Hg, Pb, Sn, Tl, V and Zn in the soil was enhanced by our biological combination. Additionally, also the organic pollutants (PAHs and PCBs) were strongly reduced up to 40.5%. About two million transcripts were identified and used for taxonomic and functional profiling. Transcripts read counts, tripartite among plant, bacteria and fungi were identified and quantified to provide an overview of the complex soil community composition. We observed that Actinobacteria and fungi abundance might be involved in remediation success. Functional analyses showed that Trehalose Biosynthesis and the antioxidant activity might have played a key-role in metaorganism effective interactions. The biotechnological approach remodeled the transcriptional profile toward organic pollutant degradation and heavy metal stress response.

Galactinol synthase 1 improves cucumber performance under cold stress by enhancing assimilate translocation.

Cucumber (Cucumis sativus L.) predominantly translocates raffinose family oligosaccharides (RFOs) in the phloem and accumulates RFOs in leaves. Galactinol synthase (GolS) catalyzes the critical step of RFO biosynthesis, and determining the functional diversity of multiple GolS isoforms in cucumber is of scientific significance. In this study, we found that all four isoforms of CsGolS in the cucumber genome were upregulated by different abiotic stresses. β-Glucuronidase staining and tissue separation experiments suggested that CsGolS1 is expressed in vascular tissues, whereas the other three CsGolSs are located in mesophyll cells. Further investigation indicates that CsGolS1 plays double roles in both assimilate loading and stress response in minor veins, which could increase the RFO concentration in the phloem sap and then improve assimilate transport under adverse conditions. Cold-induced minor vein-specific overexpression of CsGolS1 enhanced the assimilate translocation efficiency and accelerated the growth rates of sink leaves, fruits, and whole plants under cold stress. Finally, our results demonstrate a previously unknown response to adverse environments and provide a potential biotechnological strategy to improve the stress resistance of cucumber.

Advances in storage of poultry semen

Semen cryopreservation is a key biotechnological strategy used to preserve and protect genetic resources, which are subject to increasingly serious reductions in some species, and to protect animal biodiversity. Assisted reproductive techniques, however, are still not utilized to the same extent in avian species to the extent that occurs in mammals. The reasons for this situation are described in this review. The content of this paper is focused on current poultry preservation systems, published since 2010, and new strategies that are very promising for preserving avian genetic resources. Two major types of storage technologies which are utilized for avian sperm preservation, liquid storage and cryopreservation, are emphasized. The issues on which there is a focus includes supplementation of avian extenders with various compounds prior to the preservation period, use of cryoprotectants and fertility results when there were in vitro sperm evaluations. Results from recent studies indicate there are opportunities to improve the quality of bird semen after preservation. It is obvious that cryo-diluent composition may be the most important factor for development of efficacious cryopreservation methods for avian semen.

Systematic Review - State of the Art: strategies for vigor evaluation and production of high-vigor soybean seeds

Abstract: A systematic approach was employed with the objective of compiling updated and more promising information on the quality of soybean seeds, traits, treatments and analysis techniques. Among the 6,899 academic materials retrieved between 2017 and 2022, 57 of them were included in the study, separated into three groups: Group 1 - Theoretical materials addressing traits related to seed vigor, which can be used in biotechnological strategies and improvement of different species; Group 2 - Practical materials that apply techniques of treatment of soybean seeds; and Group 3 - Practical materials that apply techniques to evaluate vigor in soybean seeds. From the approach used, it was possible to observe that several genes, proteins and QTLs are associated with seed vigor. Seed treatment techniques focus on the use of biological and physical products, but there is a lack of studies validating these benefits in the field. Among the methods for evaluating the vigor of soybean seeds, the techniques of image analysis (visible light, LIBS, NIR, FTIR, FT-NIR, HSI-NIR, FHSI, Hyperspectral, Fluorescence spectral) and the use of software (Vigor-S®, SVIS®, SAPL®, IJCropSeed, Ilastik, VideometerLab®, MATLAB, ENVI®) stand out.

Overcoming the limitations of cytokines to improve cancer therapy.

Cytokines are pleiotropic soluble proteins used by immune cells to orchestrate a coordinated response against pathogens and malignancies. In cancer immunotherapy, cytokine-based drugs can be developed potentiating pro-inflammatory cytokines or blocking immunosuppressive cytokines. However, the complexity of the mechanisms of action of cytokines requires the use of biotechnological strategies to minimize systemic toxicity, while potentiating the antitumor response. Sequence mutagenesis, fusion proteins and gene therapy strategies are employed to enhance the half-life in circulation, target the desired bioactivity to the tumor microenvironment, and to optimize the therapeutic window of cytokines. In this review, we provide an overview of the different strategies currently being pursued in pre-clinical and clinical studies to make the most of cytokines for cancer immunotherapy.

Proteomic analysis reveals the metabolic versatility of Amycolatopsis sp. BX17: A strain native from milpa agroecosystem soil

Amycolatopsis sp. BX17 is an actinobacterium isolated from milpa soils, which antagonizes the phytopathogenic fungus Fusarium graminearum. Metabolites secreted by the actinobacterium cultured in glucose-free medium inhibited 100% of the mycelial growth of F. graminearum RH1, while the inhibition rate was 65% in medium supplemented with 20 g/L glucose. With the aim of studying how the metabolism of strain BX17 is modulated by glucose as the main carbon source, media with 0 and 20 g/L glucose were selected to analyze the intracellular proteins by quantitative label-free proteomic analysis. Data are available via ProteomeXchange with identifier PXD028644. Proteins identified in bacteria cultured in medium without glucose were involved in glutamate metabolism, the Krebs cycle and the shikimate pathway, suggesting that amino acids are metabolized to synthesize antifungal compounds. In glucose-containing medium, carbon flux was directed mainly toward the synthesis of energy and cell growth. This study shows the metabolic versatility of Amycolatopsis BX17, and strengthens its potential use in designing biotechnological strategies for phytopathogen control. SignificanceAmycolatopsis BX17 is a bacterium isolated from milpa agroecosystems that antagonizes the phytopathogenic fungus Fusarium graminearum. Currently, there is scarce information about the metabolism involved in the biosynthesis of antifungal agents by this genus. We used a label-free proteomic approach to identify the differences in metabolic routes for antifungal biosynthesis in Amycolatopsis BX17 grown in media with 0 and 20 g/L glucose. Taken together the results suggest that the BX17 strain could be synthesizing the antifungal metabolite(s) from the Shikimate pathway through the synthesis and degradation of the amino acid tyrosine, which is a known precursor of glycopeptides with antibiotic and antifungal activity. While the lower antifungal activity of the metabolites secreted by Amycolatopsis BX17 when grown in a medium with glucose as the main carbon source, may be correlated with a lower synthesis of antifungal compounds, due to the directing of carbon flux toward metabolic pathways involved with energy synthesis and cell growth. Likewise, it is possible that the bacteria synthesize other compounds with biological activity, such as glycopeptides with antibiotic activity. These findings are relevant because they represent the first stage to understand the metabolic regulation involved in the biosynthesis of antifungal metabolites by the genus Amycolatopsis. Finally, improving our understanding of the metabolic regulation involved in the biosynthesis of antifungal metabolites is essential to design of strategies in agricultural biotechnology for phytopathogen control.