Use Of Biotechnology Research Articles

Influence of a temperature shock on the synthesis efficiency of surfactants by <i>Rhodococcus pyridinivorans</i> 5Ap bacteria

It was found that the synthesis of surfactants by R. pyridinivorans 5Ap bacteria can be increased by exposing them to a short temperature shock (55 °C for 20 minutes) after 24 hour cultivation in a minimal medium containing molasses (3 %) and hexadecane (2 %) (9 % increase in the emulsification index). Gene activation encoding global regulators of cell metabolism, including those performing a protective function under stress, was recorded in this cultivation mode. In particular, the mRNA amount determining the synthesis of the alternative transcription factor SigH increased 90.8 times and containing in the promoters its binding sites fmdB cochaperone – 59.3 times, hsp22.5 chaperone – 81.1 times, and the trxB gene encoding thioredoxin reductase – 73.1 times. In addition, it was shown that the transcriptional activation of groEL1, groEL2 and dnaJ genes determining the synthesis of heat shock proteins was 2.2, 2.6 and 4.4 times, respectively. The obtained results suggest that an increase in the alternative factor sigma H synthesis, which activates protective cellular metabolism, as well as structural, heat shock proteins under short temperature stress, leads to an increased production of surfactants, which can be used to optimize the synthesis of these secondary metabolites for biotechnological use.

Overcoming Difficulties in Molecular Biological Analysis through a Combination of Genetic Engineering, Genome Editing, and Genome Analysis in Hexaploid Chrysanthemum morifolium.

Chrysanthemum is one of the most commercially important ornamental plants globally, of which many new varieties are produced annually. Among these new varieties, many are the result of crossbreeding, while some are the result of mutation breeding. Recent advances in gene and genome sequencing technology have raised expectations about the use of biotechnology and genome breeding to efficiently breed new varieties. However, some features of chrysanthemum complicate molecular biological analysis. For example, chrysanthemum is a hexaploid hyperploid plant with a large genome, while its genome is heterogeneous because of the difficulty of obtaining pure lines due to self-incompatibility. Despite these difficulties, an increased number of reports on transcriptome analysis in chrysanthemum have been published as a result of recent technological advances in gene sequencing, which should deepen our understanding of the properties of these plants. In this review, we discuss recent studies using gene engineering, genome editing, and genome analysis, including transcriptome analysis, to analyze chrysanthemum, as well as the current status of and future prospects for chrysanthemum.

Applications of Enzymes as Ecologic Alternatives in the Leather Industry

The aim of this paper is to study the applications of enzymes, as ecological alternatives in the leather industry. The research proposes to create ecologic leathers/furs with additional functions (non-toxic, with high-quality of soft, well-degreased surfaces) through the use of original enzyme-based biotechnologies, as new additives for leather/fur processing intended for everyday use. Another application of the enzymes studied in this paper is the development of a novel technology based on enzymes and membrane technique, for purifying wastewaters from the leather industry. We selected and analytically characterized lipases, proteases, and also new enzymes: lysozyme, tryptophan deaminase for use in leather industry. The goal was to obtain leathers/furs for everyday use, with high-performance characteristics, by processing them with enzymes – as new additives that have not been used so far in the leather industry as an alternative to the use of potentially polluting chemical materials. Complex and original technologies of ecological bioprocessing of leather/furs were used. The innovation consists in developing new biotechnologies for leather/fur with enzymatic additives in order to achieve ecologic leathers or fur articles. These biotechnologies provided advanced performances for surface quality: intense and bright colors, soft, well-degreased, and resistant to abrasion and water.

Chemical Zymogens: Specificity and Steroidal Control of Reactivation.

Activating and masking enzymatic activity on demand is of the highest importance in nature. It is achieved by chemical interconversion of enzymes and the corresponding zymogens through, for example, proteolytic processing or reversible phosphorylation, and affords on-demand activation of enzymes, controlled in space and/or time. In stark contrast, examples of chemical zymogens are very few, and in most cases these are based on disulfide chemistry, which is largely indiscriminate as to the nature of the activating thiol. In this work, we address an outstanding challenge of specificity of reactivation of chemical zymogens. We achieve this through engineering affinity between the chemical zymogen and the activator. Additional, higher-level control over zymogen reactivation is installed in a nature-mimicking approach using steroidal hormones. Taken together, the results of this study take a step towards establishing the specificity of reactivation of synthetic, chemical zymogens. We anticipate that the results of this study will contribute significantly to the development of chemical zymogens as tools for diverse use in chemical biology and biotechnology.

Phenolic compounds profile and antioxidant activity of purple passion fruit's pulp, peel and seed at different maturation stages

The purple passion fruit (Passiflora edulis Sims) is a fruit that has an intense and wrinkled purple peel when ripe. In the food industry, the passion fruit pulp is used in the production of juices and nectars, while the peel and seeds are considered waste, although they have great biotechnological potential due to their chemical composition, with emphasis on bioactive compounds. In this study, the profile of 13 phenolics, two alkaloids, the total phenolic content and the antioxidant activity, evaluated by different methods, of the pulp, peel and seed of purple passion fruit at three maturation stages, were evaluated. Fifteen metabolites in the phenolic and alkaloid profile were identified in the different fractions of the fruit, and by their variation, processes of synthesis and degradation along ripening were suggested. As for the content of phenolic compounds and antioxidant activity, the seed stood out compared to the peel and pulp. The results obtained suggest the health benefits of consuming the fruit pulp, and the potential for using the peel and seed as by-products that are also beneficial to health and as candidates for biotechnological use in industries and research centers.

Intellectual property rights in the context of biology, medicine and pharmacy: a look into the future

The article covers the study of some aspects of protecting the intellectual property rights in the areas of biology, medicine and pharmacy including biotechnological inventions. The rapid progress of science and technology, the generation and accumulation of knowledge, the possibilities to process large amounts of various data further stimulate the development of all social areas and unprecedentedly contribute to the emergence of innovations, in particular in the area of biotechnology. Intellectual property rights have a direct impact on the progress of medical and biological research, but increasingly require coordination with the ethical and moral principles of the society regarding the patentability of inventions in biology, medicine and pharmacy.The article analyzes the modern trends in the protection of intellectual property rights to biotechnology inventions at an international level, and forecasts the likely subsequent development in legal protection of intellectual property rights in the areas of biology, medicine and pharmacy in view of the ever increasing ethical and law controversies of medical and biological research in some areas, the emergence of big data, use of artificial intelligence, etc.The national legislation refers to the term «biotechnology» in the Law of Ukraine «On Environmental Protection», while not giving any legal definition of “biotechnology” in any laws. The Ukrainian intellectual property and health care legislation require reformation to get properly updated to respond to the ongoing challenges. There is a whole layer of gaps regarding the legal regulation of using biotechnology in Ukraine, and this extends to all areas where biotechnology is actually integrated. The use of biotechnology in biology, medicine and pharmacy requires a systemic and multi-aspect study in order to develop the appropriate legal norms.To ensure the stability of the pharmaceutical market, which is focused on the national manufacturer, it is necessary to create the proper conditions for the development of a high-tech research environment in Ukraine. It is necessary to improve nationallegislation in the field of biobanking and the legal regime of human biological material for the purpose of using it in research.

New insights into the carotenoid biosynthesis in Torreya grandis kernels

Torreya grandis is a characteristic rare economic tree species in subtropical mountainous areas. The nut has a variety of bioactive compounds, such as polyphenols and vitamins. However, the carotenoids compositions and its regulatory mechanisms underlying carotenoid biosynthesis in kernels remain unknown. Here, the major carotenoids in the kernel were analyzed. The result showed that lutein and β-carotene were the predominant carotenoids in the kernel, while low levels of α-carotenoid and zeaxanthin were detected. Lutein and β-carotene were decreased during the maturation of kernel. Expression analysis by RNA-seq and qRT-PCR indicated that the expressions of TgCYP97A3 and TgLCYB were also reduced during the maturation of kernel. The contents of lutein or β-carotene were obviously increased in tobacco transiently overexpressing TgCYP97A3 or TgLCYB. Moreover, DPPH radical scavenging activity and FRAP were also significantly enhanced. In addition, several MYB and WRKY transcription factors TgMYBS3, TgMYB48 and TgWRKY11 were identified to positively regulate the TgCYP97A3 expression, while TgMYB48, TgWRKY2 and TgWRKY11 could upregulate the TgLCYB expression. The illustration of carotenoids biosynthesis and its molecular mechanism in kernels not only provides a basis for understanding carotenoids biosynthesis in kernels, but also enables the use of molecular biotechnology to develop new health products rich in carotenoids based on T. grandis nuts.

Concise review on seaweed photosynthesis: From physiological bases to biotechnological applications

Seaweeds play an important role for the environment, since they are photosynthetic organisms, contributing to the cycling of nutrients and to the protection and feeding of several animals. The potential for biotechnological applications of marine macroalgae biomass is enormous, and to attend the demand of the current market, the knowledge of the physiological characteristics of these organisms is essential to ensure a high productivity in cultivation systems. In this review, we contribute to the knowledge about the photosynthetic characteristics of seaweeds, describing relevant aspects of marine macroalgae and photosynthesis, and the biotechnological use of these organisms.

Glycosylation and charge distribution orchestrates the conformational ensembles of a biotechnologically promissory phytase in different pHs – a computational study

Phytases [myo-inositol(1,2,3,4,5,6) hexakisphosphate phosphohydrolases] are phytate-specific phosphatases not present in monogastric animals. Nevertheless, they are an essential supplement to feeding such animals and for human special diets. It is crucial, hence, the biotechnological use of phytases with intrinsic stability and activity at the acid pHs from gastric environments. Here we use Metadynamics (METADY) simulations to probe the conformational space of the Aspergillus nidulans phytase and the differential effects of pH and glycosylation in this same space. The results suggest that strategic combinations of pH and glycosylation affect the stability of native-like conformations and alternate these structures from a metastable to a stable profile. Furthermore, the protein segments previously reported as more thermosensitive in phytases from this family present a pivotal role in the conformational changes at different conditions, especially H2, H5-7, L8, L10, L12, and L17. Also, the glycosylations and the pH-dependent charge balance modulate the mobility and interactions at these same regions, with consequences for the surface solvation and active site exposition. Finally, although the glycosylations have stabilized the native structure and improved the substrate docking at all the studied pHs, the data suggest a higher phytate receptivity at catalytic poses for the unglycosylated structure at pH 6.5 and the glycosylated one at pH 4.5. This behavior agrees with the exact change in optimum pH reported for this enzyme, expressed on low or high glycosylating systems. We hope the results and insights presented here will be helpful in future approaches for rational engineering of technologically promising phytases and intelligent planning of their heterologous expression systems and conditions for use. Communicated by Ramaswamy H. Sarma

Plants, Cells, Algae, and Cyanobacteria In Vitro and Cryobank Collections at the Institute of Plant Physiology, Russian Academy of Sciences-A Platform for Research and Production Center.

Ex situ collections of algae, cyanobacteria, and plant materials (cell cultures, hairy and adventitious root cultures, shoots, etc.) maintained in vitro or in liquid nitrogen (-196 °C, LN) are valuable sources of strains with unique ecological and biotechnological traits. Such collections play a vital role in bioresource conservation, science, and industry development but are rarely covered in publications. Here, we provide an overview of five genetic collections maintained at the Institute of Plant Physiology of the Russian Academy of Sciences (IPPRAS) since the 1950-1970s using in vitro and cryopreservation approaches. These collections represent different levels of plant organization, from individual cells (cell culture collection) to organs (hairy and adventitious root cultures, shoot apices) to in vitro plants. The total collection holdings comprise more than 430 strains of algae and cyanobacteria, over 200 potato clones, 117 cell cultures, and 50 strains of hairy and adventitious root cultures of medicinal and model plant species. The IPPRAS plant cryobank preserves in LN over 1000 specimens of in vitro cultures and seeds of wild and cultivated plants belonging to 457 species and 74 families. Several algae and plant cell culture strains have been adapted for cultivation in bioreactors from laboratory (5-20-L) to pilot (75-L) to semi-industrial (150-630-L) scale for the production of biomass with high nutritive or pharmacological value. Some of the strains with proven biological activities are currently used to produce cosmetics and food supplements. Here, we provide an overview of the current collections' composition and major activities, their use in research, biotechnology, and commercial application. We also highlight the most interesting studies performed with collection strains and discuss strategies for the collections' future development and exploitation in view of current trends in biotechnology and genetic resources conservation.

Fundamentals of bio-electrochemical sensing

The utilization of bio-functionalities such as biorecognition or catalysis derived them their name biosensors. Bio-electrochemical sensing is a new discipline that combines the advantages of biological detection and electrochemical transduction. Bio-electrochemical sensors are devices that use biological materials such as enzymes, antibodies, DNA, or cells as receptors to detect target analytes in a variety of samples. Electrodes convert biological interactions into electrical impulses, which can be studied using various electrochemical techniques. Bio-electrochemical sensors have demonstrated significant promise for use in clinical diagnostics, environmental monitoring, food safety, and biotechnology. Biosensors have received numerous applications in recent years because they are fast, simple, and inexpensive for practical applications. In this article, we cover the basic principles, design strategies, immobilization and regeneration techniques, along with the advantages and applications of bio-electrochemical sensors. Finally, this article discusses the rationale for developing electrochemical biosensors in the context of the various bio-receptors that can be applied.

Bacterial Toxin-Antitoxin Systems' Cross-Interactions-Implications for Practical Use in Medicine and Biotechnology.

Toxin-antitoxin (TA) systems are widely present in bacterial genomes. They consist of stable toxins and unstable antitoxins that are classified into distinct groups based on their structure and biological activity. TA systems are mostly related to mobile genetic elements and can be easily acquired through horizontal gene transfer. The ubiquity of different homologous and non-homologous TA systems within a single bacterial genome raises questions about their potential cross-interactions. Unspecific cross-talk between toxins and antitoxins of non-cognate modules may unbalance the ratio of the interacting partners and cause an increase in the free toxin level, which can be deleterious to the cell. Moreover, TA systems can be involved in broadly understood molecular networks as transcriptional regulators of other genes' expression or modulators of cellular mRNA stability. In nature, multiple copies of highly similar or identical TA systems are rather infrequent and probably represent a transition stage during evolution to complete insulation or decay of one of them. Nevertheless, several types of cross-interactions have been described in the literature to date. This implies a question of the possibility and consequences of the TA system cross-interactions, especially in the context of the practical application of the TA-based biotechnological and medical strategies, in which such TAs will be used outside their natural context, will be artificially introduced and induced in the new hosts. Thus, in this review, we discuss the prospective challenges of system cross-talks in the safety and effectiveness of TA system usage.

Multi-Stimuli-Responsive and Mechano-Actuated Biomimetic Membrane Nanopores Self-Assembled from DNA.

In bioinspired design, biological templates are mimicked in structure and function by highly controllable synthetic means. Of interest are static barrel-like nanopores that enable molecular transport across membranes for use in biosensing, sequencing, and biotechnology. However, biological ion channels offer additional functions such as dynamic changes of the entire pore shape between open and closed states, and triggering of dynamic processes with biochemical and physical stimuli. To better capture this complexity, this report presents multi-stimuli and mechano-responsive biomimetic nanopores which are created with DNA nanotechnology. The nanopores switch between open and closed states, whereby specific binding of DNA and protein molecules as stimuli locks the pores in the open state. Furthermore, the physical stimulus of high transmembrane voltage switches the pores into a closed state. In addition, the pore diameters are larger and more tunable than those of natural templates. These multi-stimuli-responsive and mechanically actuated nanopores mimic several aspects of complex biological channels yet offer easier control over pore size, shape and stimulus response. The designer pores are expected to be applied in biosensing and synthetic biology.

Recombinant Endopeptidases IdeS and IdeZ and Their Potential Application.

Endopeptidases IdeS and IdeZ (streptococcal virulence factors that specifically cleave IgG heavy chains) are of particular interest because of their potential use in biotechnology, medicine, and veterinary. Genes encoding these enzymes were cloned and expressed in Escherichia coli heterologous expression system (ideS was cloned from a Streptococcus pyogenes collection strain; ideZ from Streptococcus zooepidemicus was synthesized). The 6His-tag was introduced into the amino acid sequence of each endopeptidase, and IdeS and IdeZ were purified by metal affinity chromatography to an apparent homogeneity (according to polyacrylamide gel electrophoresis). Purified enzymes were active against human and animal IgGs; their specificity toward human IgGs was confirmed by polyacrylamide gel electrophoresis. Recombinant IdeZ was used for immunological analysis of equine strangles infection (diagnostics and determination of the titer of specific antibodies in blood). Hence, IdeZ can be used in veterinary and sanitary microbiology to diagnose infections caused by Streptococcus equi and S. zooepidemicus in addition to its application in medicine and biotechnology.

Forgotten Clones: The Birth of Cloning and the Biological Revolution. By NathanCrowe. Pittsburgh, PA: University of Pittsburgh Press. 2021. pp. 336. $60 (Hardcover).

Forgotten Clones: The Birth of Cloning and the Biological Revolution. By NathanCrowe. Pittsburgh, PA: University of Pittsburgh Press. 2021. pp. 336. $60 (Hardcover).

Associations of somatotropin (GH) gene polymorphism with lactation performance of cows

The research was conducted in the laboratory of molecular genetic expertise in 2018 and 2019 within the framework of the state assignment on the topic "Development of genetic-selection and theoretical foundations for the conservation and effective use of the gene pool of cattle in the Ural region with the use of modern biotechnologies". The object of the study is the cattle of Holsteinized black-and-white breed. The main task is to analyze the degree of influence of the somatotropin (GH) gene polymorphism on lactation performance indicators. In the course of the study, DNA from 270 cows was isolated, the polymorphism of the somatotropin gene was determined based on the obtained data, and the relationship between different genotypes and indicators of live weight, milk yield, protein mass fraction and fat mass fraction in milk was established. The results were biometrically processed using IBM SPSS Statistics 23 software program. It was revealed that LL is the main genotype of the GH gene among the representatives of the Holsteinized black-and-white breed (frequency of occurrence 77.0%). The proportion of LV and VV genotypes in the population is 21.9 and 1.1%, respectively. During the observation period, cows carrying the LL genotype showed higher milk yields in the first and third lactations. The difference with their peers was 202-2334 kg (p ≤ 0.05). A tendency towards increased protein content in milk was recorded in animals with the VV genotype. In the first lactation, the difference with LL and LV genotype carriers was 0.03 and 0.06%, respectively. In the third lactation, individuals with the VV genotype surpassed their female counterparts in protein content by 0.07 and 0.12%, respectively.

Identification and characterization of a phosphinothricin N-acetyltransferase from Enterobacter LSJC7

Phosphinothricin (PPT) is a widely used and non-selective herbicide. PPT-resistance genes, especially PPT N-acetyltransferase genes, have been used in the development of transgenic PPT-resistant crops. However, there are only a limited number of available PPT-resistance genes for use in plant biotechnology. In this study, we found that Enterobacter LSJC7 is highly resistant to PPT and can acetylate PPT to N-acetyl phosphinothricin (Ac-PPT). Furthermore, a novel PPT N-acetyltransferase gene, named LsarsN, was identified from LSJC7. When LsarsN was expressed in E. coli AW3110, it confered resistance to PPT. Ac-PPT was detected in both the culture medium and cells of AW3110 expressing the LsarsN-pET22b plasmid. The purified LsArsN protein also showed strong N-acetylation ability in vitro, and its enzymatic kinetic curve was fitted with the Michaelis-Mentan equation. Compared with wild-type LsArsN, both R72A and R74A mutants showed significantly lower PPT N-acetylation ability. In summary, our results systematically characterized LsArsN with strong ability for PPT N-acetylation, which lays the groundwork for future research into the use of this novel gene, LsarsN, to create PPT-resistant crops.

Review: Environment of the ovulatory follicle: modifications and use of biotechnologies to enhance oocyte competence and increase fertility in cattle.

The oocyte is the basis of life, supporting development from a fertilized cell to an independent multicellular organism. The oocyte's competence to drive the first cell cycles postfertilization are critical to embryonic survival and subsequent successful pregnancy. Coupled with the complex processes of follicle assembly, activation, differentiation, growth, and terminal maturation, oocyte developmental competence is gradually acquired during oocyte growth and meiotic maturation. Most reproduction management technologies and interventions are centered around these highly coordinated processes, targeting the ovarian follicle and the oocyte within. Thus, our objective was to highlight key aspects of oocyte and follicle development in cattle, and to discuss recent advances in oocyte and follicle-centered reproductive biotechnologies.

Biotechnological Potential of Hydrolytic Prokaryotic Component in Soils

The phylogenetic and functional diversity of a prokaryotic complex with biotechnological potential (carrying out the destruction of biopolymers, hydrocarbons; capable of synthesizing secondary metabolites; nitrogen fixation process) in soil and associated ecosystems has been studied. In order to identify the specifics of the development of metabolically active prokaryotes with biotechnological potential, the patterns of their distribution and the dependence of functional activity on the main environmental factors have been established. The study used molecular biological and bioinformatic approaches. The range of samples studied included modern soils (Volgograd, Tula, Moscow regions, Siberia and the Northern part of Central Kamchatka), relict habitats (Volgograd Region and Central Kamchatka) and permafrost soils of Antarctica (King George Island). The impact of anthropogenic and abiogenic loads on the development of the prokaryotic community was considered. In soils exposed to anthropogenic or abiogenic loads, along with a decrease in the diversity and abundance of prokaryotes, an increase in the number of genes marking the ability of a community to biodegradate xenobiotics, as well as genes encoding nitrogen transformations and the level of metabolism of cofactors and vitamins was found. The bacterial complex is capable of nitrification with high oil contamination of the soil, and its role also increases in the lower layers of the soil profile. Archaea play a leading role in the nitrification process in undisturbed soils. The revealed patterns indicate a high metabolic potential of the prokaryotic component of the objects under consideration and open up opportunities for biotechnological use of strains isolated from relict habitats.

New Records of Tetraselmis sp. Strains with Biotechnological Potential Isolated from Greek Coastal Lagoons

In the pursuit of sustainable sources for food, energy, and health products, microalgae have gained attention. In the present study, the lagoonal system of the Nestos River delta was selected as a sampling point in order to search for opportunistic and robust species. Two new strains of Tetraselmis are described with regards to their taxonomic features (as observed using light and transmission electron microscopy and molecular phylogenetics) and their biochemical properties (total lipid, total protein, and total carbohydrate content, photosynthetic pigments, and antioxidant capacity). The studied strains were identified as representatives of Tetraselmis verrucosa f. rubens. Furthermore, both strains exhibited an interesting biochemical profile coupled with high growth rates and promising antioxidant activity, without the use of enhancement and induction culture methods, warranting further investigation and showing potential for biotechnological use.