Biotechnological Production Research Articles

The use of a complex of microorganisms in yogurt production technology

Fermented milk products are one of the most common foods among the population. This category of products includes yogurt, which is available in its various variations: with different acidity concentrations, with various additives, and different fat content. The article presents the results of studies on the influence of a complex of microorganisms on the quality indicators of yogurt (organoleptic, physicochemical). The purpose of the research was to study the effect of different volumes of symbiotic starter on the quality indicators of yogurt. The research was carried out in the laboratory of the Department of Biotechnology and Food Products of the Ural State Agrarian University.The Methods. Four yogurt samples were produced and analyzed: a control sample and three experimental ones. The finished products were assessed for organoleptic characteristics and mass fraction of protein (GOST 31981-2013). Acidity was set according to GOST 31976-2012.The Results. According to experts, sample No. 2 was recognized as the best, with a total volume of the complex of microorganisms of 70 ml per 150 g of the finished product. This sample had a uniform consistency, undisturbed clot, and no whey separation was observed; taste and smell – pure, fermented milk; color – milky white. All test samples had deviations in the protein mass fraction. Thus, in sample No. 1, MDB was 3.4%, in samples No. 2 and No. 3 – 3.6 and 3.8%. The obtained indicators were higher than the standard values by 0.2, 0.4 and 0.6%, respectively. Acidity in all samples was within normal limits.The Conclusion. Yogurt, with a complex of microorganisms can expand the line of functional products, and the addition of symbiotic starters will improve the properties of the product and classify it as functional.

Preliminary hydrothermal treatment and steam explosion of cellulosic feedstock for the subsequent biotechnological transformation: A review

The use of renewable sources of cellulosic feedstock to produce high value-added products is a relevant issue. Cellulosic feedstock constitutes a natural matrix comprising cellulose (38–50 %), lignin (10–25 %), and hemicel-luloses (23–32 %). In order to break it down, pretreatment involving the removal of hemicelluloses and lignin is required. This process can change the chemical composition and structure of cellulose while increasing porosity. This review article provides an analysis of data on the hydrothermal treatment and steam explosion of cellulosic feedstock (sunflower straw, lawn grass, poplar sawdust, hay, reed, aspen, giant reed, silage, etc.) intended to convert it into substrates for the synthesis of biotechnological products (protein, biohydrogen, biogas, levulinic acid, methane, lactic acid, ethanol, and succinic acid). Hydrothermal treatment involves treating raw materials at 160–240 °С in water under high pressure. Pressure keeps water in a liquid state. During steam explosion, feedstock is treated with steam at a moderate temperature and pressure for a certain amount of time. Then, the pressure is rapidly released, and the fibers of cellulosic feedstock expand. The effectiveness of hydrothermal treatment and steam explosion depends both on the type of feedstock (chemical composition, solids concentration, and properties of solids) as well as on the conditions of hydrothermal treatment and steam explosion.

Review on: Public Perception of Biotechnology on Genetically Modified Crops, Bio Policy and Intellectual Property Rights

Public discussion about genetically modified crops is strongly heavily influenced by debates over their risks and benefits. Supporters of biotechnology point to its potential to reduce hunger, prevent malnutrition, treat diseases, and improve overall health and quality of life. However, there is considerable opposition to biotechnology. Some critics argue that it poses risks to human health and the environment, while others oppose it on moral and ethical grounds. The transfer of genes between different species is often criticized as "playing God" or breaking the "Law of Nature." Biosafety on (GM) crops is a rapidly growing field that includes scientific research, ethical issues, and policy and regulatory frameworks to assess and manage risks to human and animal health, including food and feed safety, as well as environmental risks related to modern biotechnology products. Bio-policy refers to the rules, norms, and ethical considerations that govern the development, production, and use of biotechnology products. These policies differ by country and can be influenced by international agreements and organizations. A key international agreement (TRIPS) Agreement, which sets global standards for intellectual property protection. The Trade-Related Aspects of Intellectual Property Rights (TRIPS) Agreement requires governments to issue patents in technological sectors, including modern biotechnology, to protect innovations in this field. This ensures that biotechnological advancements, including GM crops, are protected under intellectual property laws, aiding their development and commercialization while addressing ethical and safety concerns.

DMAD7 is a promising tool for targeted gene regulation in the methylotrophic yeast Komagataella phaffii

The methylotrophic yeast Komagataella phaffii is a popular host system for the pharmaceutical and biotechnological production of recombinant proteins. CRISPR-Cas9 and its derivative CRISPR interference (CRISPRi) offer a promising avenue to further enhance and exploit the full capabilities of this host. MAD7 and its catalytically inactive variant “dead” MAD7 (dMAD7) represent an interesting alternative to established CRISPR-Cas9 systems and are free to use for industrial and academic research. CRISPRi utilizing dMAD7 does not introduce double-strand breaks but only binds to the DNA to regulate gene expression. Here, we report the first use of dMAD7 in K. phaffii to regulate the expression of the enhanced green fluorescent protein (eGFP). A reduction of eGFP fluorescence level (up to 88 %) was achieved in random integration experiments using dMAD7 plasmids. Integration loci/events of investigated strains were assessed through whole genome sequencing. Additionally, RNA-sequencing experiments corroborated the whole genome sequencing results and showed a significantly reduced expression of eGFP in strains containing a dMAD7 plasmid, among others. Our findings conclusively demonstrate the utility of dMAD7 in K. phaffii through successfully regulating eGFP expression.

Biosynthesis of Strained Amino Acids by a PLP‐Dependent Enzyme through Cryptic Halogenation

AbstractAmino acids (AAs) are modular building blocks which nature uses to synthesize both macromolecules, such as proteins, and small molecule natural products, such as alkaloids and non‐ribosomal peptides. While the 20 main proteinogenic AAs display relatively limited side chain diversity, a wide range of non‐canonical amino acids (ncAAs) exist that are not used by the ribosome for protein synthesis, but contain a broad array of structural features and functional groups. In this communication, we report the discovery of the biosynthetic pathway for a new ncAA, pazamine, which contains a cyclopropane ring formed in two steps. In the first step, a chlorine is added onto the C4 position of lysine by a radical halogenase, PazA. The cyclopropane ring is then formed in the next step by a pyridoxal‐5′‐phosphate‐dependent enzyme, PazB, via an SN2‐like attack at C4 to eliminate chloride. Genetic studies of this pathway in the native host, Pseudomonas azotoformans, show that pazamine potentially inhibits ethylene biosynthesis in growing plants based on alterations in the root phenotype of Arabidopsis thaliana seedlings. We further show that PazB can be utilized to make an alternative cyclobutane‐containing AA. These discoveries may lead to advances in biocatalytic production of specialty chemicals and agricultural biotechnology.

Biosynthesis of Strained Amino Acids by a PLP-Dependent Enzyme through Cryptic Halogenation.

Amino acids (AAs) are modular building blocks which nature uses to synthesize both macromolecules, such as proteins, and small molecule natural products, such as alkaloids and non-ribosomal peptides. While the 20 main proteinogenic AAs display relatively limited side chain diversity, a wide range of non-canonical amino acids (ncAAs) exist that are not used by the ribosome for protein synthesis, but contain a broad array of structural features and functional groups. In this communication, we report the discovery of the biosynthetic pathway for a new ncAA, pazamine, which contains a cyclopropane ring formed in two steps. In the first step, a chlorine is added onto the C4 position of lysine by a radical halogenase, PazA. The cyclopropane ring is then formed in the next step by a pyridoxal-5'-phosphate-dependent enzyme, PazB, via an SN2-like attack at C4 to eliminate chloride. Genetic studies of this pathway in the native host, Pseudomonas azotoformans, show that pazamine potentially inhibits ethylene biosynthesis in growing plants based on alterations in the root phenotype of Arabidopsis thaliana seedlings. We further show that PazB can be utilized to make an alternative cyclobutane-containing AA. These discoveries may lead to advances in biocatalytic production of specialty chemicals and agricultural biotechnology.

Biotechnological Production of Fungal Colorants

Da eine globale Transformation in Richtung zirkulärer Bioökonomie auf nachhaltige Prozesse und umweltfreundliche Produkte angewiesen ist, werden in dieser Arbeit Alternativen für aktuell eingesetzte Farbstoffe untersucht. Die biotechnologische Produktion von Farbstoffen aus Pilzen wird als Lösungsstrategie vorgeschlagen.Zwei verschiedene Spezies wurden auf ihr Potential als Farbstoff‐Produzenten untersucht: der Gemeine Schwefelporling (Laetiporus sulphureus) und der Zottige Schillerporling (Inonotus hispidus). Beide Arten sind bekannt dafür, sowohl im Fruchtkörper, als auch im Myzel in flüssiger Nährkultur farbige Moleküle zu synthetisieren. L. sulphureus produziert die orangen Laetiporsäuren und I. hispidus bildet das gelbe Hispidin, ein bekanntes Antioxidant mit vielen weiteren Bioaktivitäten.Die Kultivierung beider Spezies wurde in dieser Arbeit vom Schüttelkolben bis zum 4 bzw. 7 L Maßstab im Bioreaktor vergrößert und die Nährmedien optimiert. Durch den Vergleich vier verschiedener L. sulphureus Stämme wurde der potenteste Produktionsstamm ausgewählt und erzielte Ausbeuten von rund 1 g/L Laetiporsäure. Bei der Kultivierung von I. hispidus wurden verschiedene physikalische und chemische Einflussfaktoren zur Steigerung der Hispidin‐Ausbeute getestet. Der Einsatz von Belichtung und oxidativem Stress stimulierte die Farbstoffsynthese, genau wie dem Nährmedium zugesetzte Prekursoren. Alle Effektoren wurden in parallelen Kultivierungen in Bioreaktoren zweier verschiedener Bauarten getestet und eine Ausbeute von 5,5 g/L Hispidin erzielt. Verschiedene Produktaufarbeitungs‐Strategien wurden etabliert, von der klassischen Extraktion bis zum Zwei‐Phasen‐System. Die zuvor geringe Stabilität der Laetiporsäure‐Extrakte konnte durch Stickstoff und Lagerung bei niedrigen Temperaturen stabilisiert werden und das Potential für die Anwendung in Textilien, Kosmetika und Lebensmitteln wurde demonstriert.Zusammenfassend wurde in dieser Arbeit anhand von zwei Beispielen gezeigt, dass die biotechnologische Produktion von Pilzfarbstoffen möglich ist. Die Maßstabsvergrößerung von Labor bis zur Pilotanlage wurde berichtet und die Kombination von chemischen und physikalischen Einflussfaktoren sorgte für Ausbeuten im g/L Bereich, die einen entsprechenden Bioprozess wettbewerbsfähig mit konventionellen Prozessen für natürliche Farbstoffe machen.

Crecimiento micelial y producción de lacasa y peroxidasas por Pleurotus ostreatus y Agrocybe aegerita en fermentación líquida

Mycelial growth of Pleurotus ostreatus and Agrocybe aegerita in malt extract broth was evaluated. Laccase, manganese peroxidase (MnP), lignin peroxidase (LiP) and unspecific peroxygenase (UnP) activities were determined in both strains, using biochemical and polyacrylamide gel electrophoresis techniques. No strain showed MnP, LiP, and UnP activities. P. ostreatus had higher biomass production and laccase yield values than those shown by A. aegerita. P. ostreatus produced approximately twice as much biomass (Xmax) compared to A. aegerita (9.12 and 5.27 g/L, respectively). Maximum laccase activity (Emax) was nine-fold higher in P. ostreatus than in A. aegerita (106.5 and 12.23 U/L, respectively). P. ostreatus and A. aegerita showed 3 bands (estimated to be approximately 42, 55, 85 and 212 kDa) and one band (estimated to be approximately 35.2 kDa) with laccase activity, respectively. These results show that laccases are constitutively produced in both fungal species, whereas MnP, LiP and UnP are inducible enzymes. P. ostreatus has a great potential in the biotechnological production of laccases.

Diskusi Mengenai Pemanfaatan Kombucha Bunga Telang Kepada Siswi SMAN 5 Cilegon Banten Sebagai Minuman Probiotik Pengendali Emosi Ketika Datang Bulan

Puberty is a transition from childhood to adolescence. Menstruation is a painful condition experienced by every woman in quite severe conditions which can result in obstacles to every daily routine. Pain during menstruation can ideally trigger an emotional increase that is quite uncontrollable, so that apart from needing spiritual self-control, physical control is needed by providing fermented biotechnology products in the form of telang flower kombucha which has been widely proven as a probiotic drink that can improve the system. immunity, inhibits microbial growth, wards off free radicals, prevents the growth of cancer cells. Teenagers who study at SMAN 5 Cilegon, especially those involved in KIR (Youth Scientific Work) activities in the field of biology, have learned a lot about the potential of butterfly pea flower kombucha in health, pharmaceutical, agricultural and livestock aspects from a series of presentations on community service materials in 2023, so that there is In collaboration with schools and biology teachers, KIR supervisors have facilitated the use of butterfly pea flower kombucha as a probiotic drink to control emotions during menstruation. This has been proven based on bivariate analysis through the results of the respective T tests both before intervention and post intervention, obtaining a p value of less than 0.005, indicating that H0 is rejected while Ha is accepted, so there is a significant influence on telang flower kombucha after being given in controlling emotions when period.

Visualizing Metabolism in Biotechnologically Important Yeasts with dDNP NMR Reveals Evolutionary Strategies and Glycolytic Logic.

Saccharomyces cerevisiae has long been a pillar of biotechnological production and basic research. More recently, strides to exploit the functional repertoire of nonconventional yeasts for biotechnological production have been made. Genomes and genetic tools for these yeasts are not always available, and yeast genomics alone may be insufficient to determine the functional features in yeast metabolism. Hence, functional assays of metabolism, ideally in the living cell, are best suited to characterize the cellular biochemistry of such yeasts. Advanced in cell NMR methods can allow the direct observation of carbohydrate influx into central metabolism on a seconds time scale: dDNP NMR spectroscopy temporarily enhances the nuclear spin polarization of substrates by more than 4 orders of magnitude prior to functional assays probing central metabolism. We use various dDNP enhanced carbohydrates for in-cell NMR to compare the metabolism of S. cerevisiae and nonconventional yeasts, with an emphasis on the wine yeast Hanseniaspora uvarum. In-cell observations indicated more rapid exhaustion of free cytosolic NAD+ in H. uvarum and alternative routes for pyruvate conversion, in particular, rapid amination to alanine. In-cell observations indicated that S. cerevisiae outcompetes other biotechnologically relevant yeasts by rapid ethanol formation due to the efficient adaptation of cofactor pools and the removal of competing reactions from the cytosol. By contrast, other yeasts were better poised to use redox neutral processes that avoided CO2-emission. Beyond visualizing the different cellular strategies for arriving at redox neutral end points, in-cell dDNP NMR probing showed that glycolytic logic is more conserved: nontoxic precursors of cellular building blocks formed high-population intermediates in the influx of glucose into the central metabolism of eight different biotechnologically important yeasts. Unsupervised clustering validated that the observation of rapid intracellular chemistry is a viable means to functionally classify biotechnologically important organisms.

Application of fed-batch strategy to fully eliminate the negative effect of lignocellulose-derived inhibitors in ABE fermentation

BackgroundInhibitors that are released from lignocellulose biomass during its treatment represent one of the major bottlenecks hindering its massive utilization in the biotechnological production of chemicals. This study demonstrates that negative effect of inhibitors can be mitigated by proper feeding strategy. Both, crude undetoxified lignocellulose hydrolysate and complex medium supplemented with corresponding inhibitors were tested in acetone–butanol–ethanol (ABE) fermentation using Clostridium beijerinckii NRRL B-598 as the producer strain.ResultsFirst, it was found that the sensitivity of C. beijerinckii to inhibitors varied with different growth stages, being the most significant during the early acidogenic phase and less pronounced during late acidogenesis and early solventogenesis. Thus, a fed-batch regime with three feeding schemes was tested for toxic hydrolysate (no growth in batch mode was observed). The best results were obtained when the feeding of an otherwise toxic hydrolysate was initiated close to the metabolic switch, resulting in stable and high ABE production. Complete utilization of glucose, and up to 88% of xylose, were obtained. The most abundant inhibitors present in the alkaline wheat straw hydrolysate were ferulic and coumaric acids; both phenolic acids were efficiently detoxified by the intrinsic metabolic activity of clostridia during the early stages of cultivation as well as during the feeding period, thus preventing their accumulation. Finally, the best feeding strategy was verified using a TYA culture medium supplemented with both inhibitors, resulting in 500% increase in butanol titer over control batch cultivation in which inhibitors were added prior to inoculation.ConclusionProperly timed sequential feeding effectively prevented acid-crash and enabled utilization of otherwise toxic substrate. This study unequivocally demonstrates that an appropriate biotechnological process control strategy can fully eliminate the negative effects of lignocellulose-derived inhibitors.Graphical

Antioxidant Activity in Pharmaceutical Biotechnology Products in The Form of Formulations and Preparations of Telang Flower Kombucha Hand Soap (Clitoria ternatea L)

Free radicals need to be handled and even prevented so that they are not exposed to the skin, especially the hands. One solution to preventing the exposure of free radicals to the skin of the hands is to wash your hands with herbal soap which is made from the active ingredient of telang flower kombucha. Telang flower kombucha was designed as an active ingredient in the formulation and hand washing soap preparation in this research has been proven as a pharmaceutical biotechnology product in warding off free radicals. This research was designed by making a formulation and preparation of telang flower kombucha hand washing soap at concentrations of 20%, 30% and 40% then measuring it using UV-Vis spectrophotometry. Based on statistical analysis, especially post hoc analysis of pharmaceutical biotechnology products in the form of formulations and preparations of telang flower kombucha hand washing soap at a concentration of 40%, it is significantly different from 20% and 30% as a source of antioxidants.

Engineering Bifunctional Galactokinase/Uridyltransferase Chimera for Enhanced UDP-d-Xylose Production.

The biotechnological production of uridine diphosphate-d-xylose (UDP-d-xylose), the glycosyl donor in enzymatic for d-xylose, is an important precursor for advancing glycoengineering research on biopharmaceuticals such as heparin and glycosaminoglycans. Leveraging a recently discovered UDP-xylose salvage pathway, we have engineered a series of bifunctional chimeric biocatalysts derived from Solitalea canadensis galactokinase/uridyltransferase, facilitating the conversion of d-xylose to UDP-d-xylose. This study elucidates the novel assembly of eight fusion protein constructs, differing in domain orientations and linker peptide lengths, to investigate their functional expression in Escherichia coli, resulting in the synthesis of the first bifunctional enzyme that orchestrates a direct transformation from d-xylose to UDP-d-xylose. Fusion constructs with a NH2-GSGGGSGHM-COOH peptide linker demonstrated the highest expression and catalytic tenacity. For the highest catalytic conversion from d-xylose to UDP-d-xylose, we established an optimum pH of 7.0 and a temperature optimum of 30 °C, with an optimal fusion enzyme concentration of 3.3 mg/mL for large-scale UDP-d-xylose production. Insights into ATP and ADP inhibition further helped to optimize the reaction conditions. Testing various ratios of unfused galactokinase and uridyltransferase biocatalysts for UDP-xylose synthesis from d-xylose revealed that a 1:1 ratio was optimal. The K cat/K m value for the NH2-GSGGGSGHM-COOH peptide linker showed a 10% improvement compared with the unfused counterparts. The strategic design of these fusion enzymes efficiently routes for the convenient and efficient biocatalytic synthesis of xylosides in biotechnological and pharmaceutical applications.

Screening of Probiotic Candidates Bacteria as Biocontrol of Aeromonas Hydrophila Pathogen Isolated from Mina Padi Cultivation Area

The agricultural activities in rice-farming system can have an impact on fish farming and the characteristics of bacterial community, especially pathogenic bacteria. Utilization of probiotics as environmentally friendly biotechnology products can be used to improve environmental quality and suppress the presence of pathogenic bacteria. This study aimed to select bacteria as probiotic agents from aquaculture ponds with the Rice-fish farming system. A total of 22 bacterial isolates were isolated from the water and sediment contained in the culture ponds. Based on the screening results, 15 isolates were confirmed as general non-pathogenic bacteria (Aeromonas sp.), 9 isolates had antibacterial activity against Aeromonas hyrophylla and 4 isolates showed high antibiotic sensitivity and were able to synergize. The results showed the Proteus mirabilis, Proteus penneri, Kurthia gibsonii and Bacillus cereus strains. Bacillus cereus strain LB8 has antibacterial activity that can inhibit the pathogen Aeromonas hydrophila with an inhibition zone of 8mm and has a very high sensitivity to antibiotics. These four isolates are able to work together synergistically and can be used as consortium probiotic bacterial agents to suppress the growth of pathogens.

Bioproduction of xylitol by Candida tropicalis 13803 from pistachio shell hydrolysate obtained through MW-HPCO2 system

Objective: Biotechnological xylitol production from secondary agricultural residues is a promising approach for a sustainable and environmental purpose. Lignocellulosic biomass is a significant feedstock for biofuel and biochemical production. Its accessibility, cost-effectiveness, renewability, and environmental friendliness make it an attractive alternative to fossil fuels and other conventional sources of energy and chemicals. Materials and methods: In this study, the conversion of xylan to xylose in a pistachio shell was provided with a novel technology of a microwave-assisted high-pressure CO2/H2O system. Xylose rich pistachio shell hydrolysate was utilized by Candida tropicalis ATCC 13803 for xylitol production. Different concentrations of xylose (50, 100, and 150 g/L) were employed for xylitol production in shake-flask. Results and conclusion: HMF and furfural were completely removed from xylose-rich hydrolysate by activated charcoal. The improvement in yeast performance was limited with increasing xylose concentration. The highest xylitol produced by C. tropicalis from pistachio shell hydrolysate (65.15 g/L) and the maximum yield of xylitol 0.66 g/g with 100 g/L xylose were obtained in shake-flask whereas xylitol produced at 50 g/L and 150 g/L xylose were 0.65 and 0.37 g/g, respectively. Volumetric productivity at 100 g/L of xylose was 1.28 times and 1.84 times higher compared to xylose concentrations of 50 g/L and 150 g/L, respectively. Xylitol production performance (71.73 g/L) of detoxified pistachio shell hydrolysate at 100 g/L of xylose was almost identical to pure xylose. However, the yeast was not able to consume xylose at 150 g/L resulting in no xylitol production.

Natural biosynthesis, pharmacological applications, and sustainable biotechnological production of ornamental plant-derived anthocyanin: beyond colorants and aesthetics.

Flowers have long been admired for their aesthetic qualities and have even found their way to be included in the human diet. Among the many chemical compounds found in flowers, anthocyanins stand out for their versatile applications in the food, cosmetic, and nutraceutical industries. The biosynthetic pathway of anthocyanins has been thoroughly studied in certain flower species, leading to the detection of key regulatory genes that can be controlled to enhance the production of anthocyanins via biotechnological methods. Nevertheless, the quantity and form of anthocyanins found in natural sources differ, bothqualitativelyandquantitatively, depending on the ornamental plant species. For this reason, research on in vitroplantcultures has been conducted for years in an attempt to comprehend how these essential substances are produced. Different biotechnological systems, like in vitro plant cell, organ, and tissue cultures, and transgenic approaches, have been employed to produce anthocyanins under controlled conditions. However, multiple factors influence the production of anthocyanins and create challenges during large-scale production. Metabolic engineering techniques have also been utilized for anthocyanin production in microorganisms and recombinant plants. Although these techniques are primarily tested at lab- and pilot-scale, limited studies have focused on scaling up the production. This review analyses the chemistry and biosynthesis of anthocyanin along with the factors that influence the biosynthetic pathway. Further emphasis has been given on strategies for conventional and non-conventional anthocyanin production along with their quantification, addressing the prevailing challenges, and exploring ways to ameliorate the production using the in vitro plant cell and tissue culture systems and metabolic engineering to open up new possibilities for the cosmetic, pharmaceutical, and food industries.

Induction of point and structural mutations in engineered yeast Saccharomyces cerevisiae improve carotenoid production.

β-Carotene is an attractive compound and that its biotechnological production can be achieved by using engineered Saccharomyces cerevisiae. In a previous study, we developed a technique for the efficient establishment of diverse mutants through the introduction of point and structural mutations into the yeast genome. In this study, we aimed to improve β-carotene production by applying this mutagenesis technique to S. cerevisiae strain that had been genetically engineered for β-carotene production. Point and structural mutations were introduced into β-carotene-producing engineered yeast. The resulting mutants showed higher β-carotene production capacity than the parental strain. The top-performing mutant, HP100_74, produced 37.6mg/L of β-carotene, a value 1.9 times higher than that of the parental strain (20.1mg/L). Gene expression analysis confirmed an increased expression of multiple genes in the glycolysis, mevalonate, and β-carotene synthesis pathways. In contrast, expression of ERG9, which functions in the ergosterol pathway competing with β-carotene production, was decreased in the mutant strain. The introduction of point and structural mutations represents a simple yet effective method for achieving mutagenesis in yeasts. This technique is expected to be widely applied in the future to produce chemicals via metabolic engineering of S. cerevisiae.

PHBV cycle of life using waste as a starting point: from production to recyclability

Global concern about plastic pollution is forcing new policies and modifications of human consumption as well as promoting new research lines aiming at the replacement of non-degradable plastics with other polymers more environmentally friendly. Addressing food waste and promoting circular economy strategies, among other approaches, are crucial in reducing environmental impacts and fostering sustainability in several sectors like the agri-food industry. The European Union’s Circular Economy Action Plan is a significant initiative in this direction. Biotechnological processes, especially the valorisation of agri-food waste to produce highly marketed biomolecules like poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) using microorganisms as cellular factories, offer promising avenues for achieving these goals. PHBV is a biodegradable polymer firstly characterised as an isolated biopolymer from bacterial biomass. This biopolymer shows interesting physicochemical properties making possible immense potential in various applications due to its biocompatibility and sustainability, thus revealing it as a good candidate to replace plastics produced by chemical synthesis from petroleum (which are highly recalcitrant and consequently pollutants). This review critically analyses the PHBV synthesis and end-of-life scenarios from their synthesis using chemical and biological pathways, through the forms of biotechnological operation and production, to the forms described until the moment of recycling.

Isolation and characterization of a phage collection against Pseudomonas putida.

The environmental bacterium, Pseudomonas putida, possesses a broad spectrum of metabolic pathways. This makes it highly promising for use in biotechnological production as a cell factory, as well as in bioremediation strategies to degrade various aromatic pollutants. For P. putida to flourish in its environment, it must withstand the continuous threats posed by bacteriophages. Interestingly, until now, only a handful of phages have been isolated for the commonly used laboratory strain, P. putida KT2440, and no phage defence mechanisms have been characterized. In this study, we present a new Collection of Environmental P. putida Phages from Estonia, or CEPEST. This collection comprises 67 double-stranded DNA phages, which belong to 22 phage species and 9 phage genera. Our findings reveal that most phages in the CEPEST collection are more infectious at lower temperatures, have a narrow host range, and require an intact lipopolysaccharide for P. putida infection. Furthermore, we show that cryptic prophages present in the P. putida chromosome provide strong protection against the infection of many phages. However, the chromosomal toxin-antitoxin systems do not play a role in the phage defence of P. putida. This research provides valuable insights into the interactions between P. putida and bacteriophages, which could have significant implications for biotechnological and environmental applications.

Distribution and Level of Bioactive Monoacylglycerols in 12 Marine Microalgal Species.

Microalgae are currently considered an attractive source of highly valuable metabolites potentially exploitable as anticancer agents, nutraceuticals and cosmeceuticals and for bioenergy purposes. Their ease of culturing and their high growth rates further promote their use as raw material for the production of specialty products. In the present paper, we focused our attention on specific glycerol-based lipid compounds, monoacylglycerols (MAGs), which displayed in our previous studies a selective cytotoxic activity against the haematological U-937 and the colon HCT-116 cancer cell lines. Here, we performed a quali/quantitative analysis of MAGs and total fatty acids (FAs) along with a profiling of the main lipid classes in a panel of 12 microalgal species, including diatoms and dinoflagellates. Our results highlight an inter- and intraspecific variability of MAG profile in the selected strains. Among them, Skeletonema marinoi (strain FE7) has emerged as the most promising source for possible biotechnological production of MAGs.