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Vol 61, No 5 (2025)
Articles
Microwave sensor system for biologically significant antigens detection (review)
Abstract
The possibility of combining recognizing (sensitive) biological components with various converters creates a huge variety of biosensor systems. A promising direction is to develop biosensors based on microwave resonators. The operation of microwave sensors is based on a change in the resonant frequency of an electromagnetic resonator upon contact with the biological object under study . This review shows the possibilities of microwave sensor systems and discusses the various analytic options in the determination of biomolecules by using antibiotics and disease markers, including glucose and antibodies, as examples. The presented results show the promise of microwave sensor systems for biomolecule determination and for their further use in biomedicine.
Applied Biochemistry and Microbiology. 2025;61(5):443-457
443-457
The Role of Adenylate Cyclase and сAMP in Controlling the Virulence of Animal Bacterial Pathogensrole Phytopathogens and Plant Mutuals
Abstract
According to the information available today, all types of microorganisms have common mechanisms for regulating the activity of virulence factors by the secondary messenger cAMP. They have been best studied in human and animal pathogens. At the same time, microorganisms that differ in specialization and habitat conditions, such as phytopathogens and mutualists, have mechanisms controlled by cAMP and adenylate cyclases that are fundamentally different from those in animal pathogens. The level of study of these processes in microorganisms of different specializations is uneven. The review attempts to systematize the available literature data and conduct a comparative analysis.
Applied Biochemistry and Microbiology. 2025;61(5):458-467
458-467
Glycation of leghemoglobin by methylglyoxal in comparison with other hemoglobins and influence on their peroxidase activity
Abstract
Non-enzymatic glycation is an irreversible posttranslational pro tein modification, which leads to a violation of physico-chemical properties and functions. Glycation most often affects lysine and arginine residues. Since hemoglobins contain many lysine residues (average 9%), they are often target s for glycating agents glyoxal and methylglyoxal (MG). A comparative study of the susceptibility for glycation of leghemoglobin (Lb) from bean nodules (Vicia faba L.), myoglobins (Mb) from sperm whale muscles and horse heart, and hemoglobins (Hb) from bovine and human erythrocytes was carried out. The level of glycation was defined by the autofluorescence of protein-bound advanced glycation end products (AGEs). The glycation level of Lb was 2.5 times higher than of sperm whale Mb and human Hb and 5 times higher than of horse Mb and bovine Hb. Lb glycation level depended on the presence of oxygen in the medium. Under microaerobic conditions, amount of AGEs formed was 3 times lower than in oxygen-containing environment, and the degradation of heme group was also slower. Glycation also affected the peroxidase activity of hemoproteins. Initial rate of Lb peroxidase reaction was 6 times higher than of myoglobins and 10–13 times higher than of hemoglobins. Glycation decreased the rate of Lb and hemoglobins peroxidase reaction, while for myoglobins it did not change or increased depending on incubation time with MG.
Applied Biochemistry and Microbiology. 2025;61(5):468-477
468-477
Impact of Neurotransmitters on the Photosynthetic Pigment Content of the Green Microalga Haematococcus lacustris (Strains IPPAS H-239 and BM-1).
Abstract
The present work deals with the effects of the neurotransmitters serotonin, histamine, dopamine, norepinephrine, and acetylcholine at concentrations of 0.1—10 m M on chlorophyll and carotenoid contents of Haematococcus lacustris strains IPPAS H-239 and BM-1. In strain H. lacustris BM-1, all tested neurotransmitters except serotonin were found to stimulate carotenoid formation with an increase in carotenoid content in the cells. The stimulatory effect was quite significant with acetylcholine and especially histamine and less manifest with dopamine and norepinephrine. Carotenoid formation by strain IPPAS H-239 was only stimulated by acetylcholine and, to a lesser extent, by norepinephrine. The other neurotransmitters inhibited carotenoid formation. The total chlorophyll a and b content increased in the presence of all tested neurotransmitters except serotonin in strain BM-1. As for strain IPPAS H-239, its chlorophyll content was increased by acetylcholine and norepinephrine, whereas histamine and serotonin lowered the chlorophyll content. It is suggested that the tested neurotransmitters influence the vegetative cell – palmelloid cell – encysted cell transition, fixing it at the intermediate brown palmelloid stage characterized by significant chlorophyll and carotenoid contents.
Applied Biochemistry and Microbiology. 2025;61(5):478-486
478-486
Optimization of transformation conditions by electroporation for Mycobacterium abscessus
Abstract
Efficient transformation of mycobacteria, in particular M. abscessus , is significantly complicated by the specific structure of their cell wall. The most widely used and effective method of introducing plasmid and phage DNA into mycobacterial cells is electroporation. The efficiency of electroporation is significantly affected by many factors, such as the nature of the DNA, the selective marker, growth supplements, the parameters of the electrical impulse, the species and the strain of the recipient mycobacterium. Although conditions for efficient electroporation for the slow-growing pathogen M. tuberculosis and the fast-growing saprophyte M. smegmatis have been described in details, recommendations for M. abscessus are scattered and even contradictory. Here it was established that efficient transformation of M. abscessus ATCC 19977 with the replicative vector pMV261 by electroporation is possible when using a logarithmic growth phase culture in a fairly wide range of optical density values OD 600 = 0.8–4.2, while cooling has little effect on the transformation frequency. A critical parameter is the mass of the introduced DNA. It has been established that the number of transformants obtained per 1 µg of DNA increases proportionally to the square of its mass. In case of introducing less than 0.5 μ g of plasmid DNA the efficiency of electroporation is insufficient.
Applied Biochemistry and Microbiology. 2025;61(5):487-493
487-493
Antimicrobial Potential Micromycete Emericellopsis sp. E102 and the Influence of Cultivation Conditions on the Biosynthesis of Antibiotics
Abstract
In recent decades, the interest in Emericellopsis genus as producers of bioactive molecules has increased significantly due to the isolation of new compounds with potential pharmaceutical applications. Evaluation of the spectrum of antibiotic activity has allowed us to choose a promising producer of new antibacterial compound, Emericellopsis sp. E102 strain derived from saline soils. Strain E102, based on molecular and phylogenetic constructions, is allocated to a separate clade within the marine clade of Emericellopsis and is presumably a new species. The ethyl acetate extract of E102 strain demonstrated significant efficacy in a concentration of 1.000 μ g/mL, resulting in inhibition zones measuring 20.3– 30.0 mm against Escherichia coli ATCC 25922; Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 700603, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212. The high-performance liquid chromatography analysis identified compound with monoisotopic mass of the substance 724.5 g/mol. Using the Chemcalc.org service, the most probable gross formulas of the required component were determined. Based on the presented calculations, there is a high probability that the substance has a sterane framework.
Applied Biochemistry and Microbiology. 2025;61(5):494-503
494-503
Study of Localization of Endogenous N-Phenyl-2-Naphthylamine and Phthalates in Pea Seedlings (Pisum sativum L.)
Abstract
The content of negative allelopathic substances, N-phenyl-2-naphthylamine ( N-PNA ) and phthalates, was studied in morphologically and physiologically different root sites, in the epicotile of etiolated pea seedlings ( Pisum sativum L.) and in the callus obtained from their epicotiles. The detection of N-PNA, o -phthalic acid, and phthalates in callus cells is evidence of the independent biosynthesis of these compounds in pea plant cells. Based on the changes in the content of N-PNA and phthalates caused by 10 –4 M naphthalene, it was shown that the main loci of biosynthesis of these compounds in pea seedlings include the growing apical part of the root and the epicotyle. It has been suggested that legumes, characterized by the ability to metabolize these substances with the participation of endophytic bacteria capable of degrading PAHs, can be used for remediation of soils contaminated with PAH pollutants.
Applied Biochemistry and Microbiology. 2025;61(5):504-510
504-510
Low Temperatures Stimulate Alternative Splicing of the CPK26 Gene in Vitis amurensis Grapes
Abstract
Alternative splicing ( AS ) is a non-canonical gene splicing process that allows a single gene to synthesise multiple protein isoforms and enhance a variety of protein functions. In this study, the involvement of AS in the generation of plant resistance to abiotic stresses was investigated using the VaCPK26 calcium-dependent protein kinase ( CPK ) gene, which is responsible for the resistance of Vitis amurensis Rupr. grapes to soil salinity and drought. The level of VaCPK26 transcription in grape leaves was studied under the influence of different environmental factors. Under low temperature exposure, in addition to the full-length VaCPK26 transcript, a short-spliced VaCPK26s1 transcript was obtained that lacked the 2nd exon of the 7 that make up the full-length VaCPK26 . Recombinant VaCPK26 increased the resistance of grape cells to salt stress and drought, and overexpression of the spliced VaCPK26s1 transcript in V. amurensis grape cell cultures had no effect on resistance to the stresses tested. These results show that AS can lead to the loss of properties of spliced transcripts characteristic of the original full-length form, which is important for a complete understanding of the biological functions of CPK and alternative splicing.
Applied Biochemistry and Microbiology. 2025;61(5):511-520
511-520
Competitive Polarization Fluorescence Aptamer Analysis of Aflatoxin B1: Influence of Salt Composition of the Reaction Medium on the Interaction of Reagents and Detection Limit
Abstract
The work examines the interaction of an aptamer (a synthetic receptor based on single-stranded DNA) having a loop structure with a specific ligand – aflatoxin B1 (AFB1), a common toxic contaminant of food products. The effects of mono- and divalent cations (Cs +, Li + , Na + , K +, Sr 2+, Ba 2+ , Mg 2+, Ca 2+) in different concentrations on the reactivity of the aptamer were studied. The fluorescence anisotropy (FA) of the fluorescein-labeled AFB1 derivative was recorded, reflecting both binding to the aptamer and the mobility of the aptamer – labeled AFB1 complex in a given reaction medium. It was shown that the recorded changes in FA differ significantly depending on the cation present in the reaction medium and its concentration. The effect on FA of monovalent cations was observed in the range of 400 mM–2.5 M, for divalent cations it is more pronounced and corresponds to concentrations from 6 to 200 mM. According to the degree of FA changes, the cations form the rows Cs + << K + ≤ ≤ Li + < Na + and Ba 2+ < Sr 2+ < Ca 2+ < Mg 2+. Comparison of the characteristics of the competitive determination of AFB1 for the traditional reaction medium and the medium with a 50-fold (from 20 mM to 1.0 M) increase in the concentration of magnesium acetate demonstrated that the given increase caused a 12-fold decrease in the detection limit – to 2.5 ± 0.4 nM. The results obtained allow considering choice of cation content as an effective tool for creating highly sensitive aptamer-based analytical systems.
Applied Biochemistry and Microbiology. 2025;61(5):521-528
521-528