Pub Date : 2024-07-27DOI: 10.1134/s0003683824604384
A. A. Klimova, I. V. Grigoriev, D. V. Vasina, M. N. Anurova, V. A. Gushchin, N. P. Antonova
Abstract
In recent years modified bacteriophage lysins have widely been investigated for the purposes of development of antibacterial therapy. Thus, effective and precise methods for the quantitative analysis of these enzymes are in high demand. The enzyme-linked immunosorbent assay (ELISA) method has been developed for the detection of recombinant modified endolysin LysAm24-SMAP in biological samples. The optimal parameters for protein detection were determined, in particular, the influence of salt and the composition of the buffer system for preparation of the samples was studied. The applicability of the immunodetection system of the genetically engineered endolysin LysAm24-SMAP in various biological samples with enzyme concentrations from 0.4 ng/mL was demonstrated. In addition, the influence of matrix effects in samples of animal organs and tissue homogenates and producer strain lysates and their individual components during the analysis was assessed and it was shown that 0.65 M NaCl addition in the ELISA buffer is crucial for achieving correct results and reduces nonspecific interactions in the case of LysAm24-SMAP. The effectiveness of the developed system in the immunochemical control of the bacteriolytic enzyme was confirmed.
{"title":"Development of Immunoassay for Detection of Engineered Endolysin LysAm24-SMAP","authors":"A. A. Klimova, I. V. Grigoriev, D. V. Vasina, M. N. Anurova, V. A. Gushchin, N. P. Antonova","doi":"10.1134/s0003683824604384","DOIUrl":"https://doi.org/10.1134/s0003683824604384","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>In recent years modified bacteriophage lysins have widely been investigated for the purposes of development of antibacterial therapy. Thus, effective and precise methods for the quantitative analysis of these enzymes are in high demand. The enzyme-linked immunosorbent assay (ELISA) method has been developed for the detection of recombinant modified endolysin LysAm24-SMAP in biological samples. The optimal parameters for protein detection were determined, in particular, the influence of salt and the composition of the buffer system for preparation of the samples was studied. The applicability of the immunodetection system of the genetically engineered endolysin LysAm24-SMAP in various biological samples with enzyme concentrations from 0.4 ng/mL was demonstrated. In addition, the influence of matrix effects in samples of animal organs and tissue homogenates and producer strain lysates and their individual components during the analysis was assessed and it was shown that 0.65 M NaCl addition in the ELISA buffer is crucial for achieving correct results and reduces nonspecific interactions in the case of LysAm24-SMAP. The effectiveness of the developed system in the immunochemical control of the bacteriolytic enzyme was confirmed.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141782831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604414
L. R. Khakimova, O. V. Chubukova, Z. R. Vershinina
Abstract—
The results of study of the effects of the Pseudomonas sp. OBA 2.4.1 PGPB strain resistant to NiCl2 (up to 3 mM), Pb(CH3COO)2 (up to 5 mM), and glyphosate (up to 8 mg/mL) on Pisum sativum L. plants at different concentrations of the heavy metals and the herbicide are reported. It was found that this strain had a positive effect on the root length of pea seedlings in the presence of heavy metals indicating an increase in the plant resistance to nickel and lead stress. However, no such effect was observed in the experiment with glyphosate, which confirmed the high toxicity of this compound. The results obtained demonstrated that the Pseudomonas sp. OBA 2.4.1 strain promoted the growth of Pisum sativum L. under nickel and lead stress; these findings can further be used for the development of complex biopreparations intended both for the protection of agricultural plants from the effects of heavy metals and for remediation of contaminated soils.
{"title":"Use of the Pseudomonas sp. OBA 2.4.1 Strain for Presowing Treatment of Pea Seeds (Pisum Sativum L.) in the Presence of Heavy Metals and Glyphosate","authors":"L. R. Khakimova, O. V. Chubukova, Z. R. Vershinina","doi":"10.1134/s0003683824604414","DOIUrl":"https://doi.org/10.1134/s0003683824604414","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">\u0000<b>Abstract</b>—</h3><p>The results of study of the effects of the <i>Pseudomonas</i> sp. OBA 2.4.1 PGPB strain resistant to NiCl<sub>2</sub> (up to 3 mM), Pb(CH<sub>3</sub>COO)<sub>2</sub> (up to 5 mM), and glyphosate (up to 8 mg/mL) on <i>Pisum sativum</i> L. plants at different concentrations of the heavy metals and the herbicide are reported. It was found that this strain had a positive effect on the root length of pea seedlings in the presence of heavy metals indicating an increase in the plant resistance to nickel and lead stress. However, no such effect was observed in the experiment with glyphosate, which confirmed the high toxicity of this compound. The results obtained demonstrated that the <i>Pseudomonas</i> sp. OBA 2.4.1 strain promoted the growth of <i>Pisum sativum</i> L. under nickel and lead stress; these findings can further be used for the development of complex biopreparations intended both for the protection of agricultural plants from the effects of heavy metals and for remediation of contaminated soils.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141786243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604438
A. A. Byvalov, L. G. Dudina, T. B. Kravchenko, S. A. Ivanov, I. V. Konyshev, N. A. Morozova, A. V. Chernyadiev, S. V. Dentovskaya
Abstract
The role of surface antigens of Yersinia pestis in reception of the phage L-413C was evaluated experimentally. Based on the methods of phage inactivation after its co-incubation with soluble or bead-bounded antigens, the importance of the plague microbe LPS in the phage reception was confirmed, and the inability to bind the capsular antigen F1, Ail protein, and two autotransporters YapF and YapM was shown. The native and recombinant PsaA, being solved, significantly inhibited the lytic activity of the phage in contrast to the bead-bound antigens. The knockout EV cells (ΔpsaA) are able to bind the phage particles as well as the wild strain. The use of three methods to evaluate the role of the PsaA antigen in phage L-413C reception gave contradictory results. On the one hand, the reactive domains of PsaA are able to interact with phage particles in solution. At the same time, these domains appear to determine nonspecific binding of the PsaA protein to the underlying bacterial cell structures or polystyrene microsphere, preventing phage adhesion.
{"title":"The Role of Yersinia pestis Antigens in the Reception of Plague Diagnostic Bacteriophage L-413C","authors":"A. A. Byvalov, L. G. Dudina, T. B. Kravchenko, S. A. Ivanov, I. V. Konyshev, N. A. Morozova, A. V. Chernyadiev, S. V. Dentovskaya","doi":"10.1134/s0003683824604438","DOIUrl":"https://doi.org/10.1134/s0003683824604438","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The role of surface antigens of <i>Yersinia pestis</i> in reception of the phage L-413C was evaluated experimentally. Based on the methods of phage inactivation after its co-incubation with soluble or bead-bounded antigens, the importance of the plague microbe LPS in the phage reception was confirmed, and the inability to bind the capsular antigen F1, Ail protein, and two autotransporters YapF and YapM was shown. The native and recombinant PsaA, being solved, significantly inhibited the lytic activity of the phage in contrast to the bead-bound antigens. The knockout EV cells (Δ<i>psaA</i>) are able to bind the phage particles as well as the wild strain. The use of three methods to evaluate the role of the PsaA antigen in phage L-413C reception gave contradictory results. On the one hand, the reactive domains of PsaA are able to interact with phage particles in solution. At the same time, these domains appear to determine nonspecific binding of the PsaA protein to the underlying bacterial cell structures or polystyrene microsphere, preventing phage adhesion.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141783001","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604360
A. Yu. Gulevich, A. Yu. Skorokhodova, V. G. Debabov
Abstract
The biosynthesis of butyric acid from glucose through inverted fatty acid β-oxidation by recombinant Escherichia coli strains was optimized. An increased yield of the target compound was achieved resulting from the plasmid expression of the atoB, fadB, and fadE/fabI genes in the core strain MG∆4 PL-tesB ΔyciA (MG1655 ∆ackA-pta, ∆poxB, ∆ldhA, ∆adhE, PL-SDϕ10-tesB, ∆yciA). The positive effect of enforced ATP hydrolysis on microaerobic conversion of the carbohydrate substrate to the final product by the recombinants was demonstrated. Activation of the futile cycle of pyruvate–phosphoenolpyruvate–pyruvate, due to the increased expression of the ppsA gene, ensured a marked increase in glucose consumption by the recombinants and led to an increase in the molar yield of butyric acid up to 39.5%. When the components of the H+-ATP synthase complex were uncoupled resulting from the deletion of atpFH genes, the molar yield of butyric acid from glucose demonstrated by the strain forming butyryl-CoA by the action of enoyl-ACP reductase FabI reached 46%.
{"title":"Optimization of the Biosynthesis of Butyric Acid from Glucose through the Inverted Fatty Acid β-Oxidation Pathway by Recombinant Escherichia coli Strains","authors":"A. Yu. Gulevich, A. Yu. Skorokhodova, V. G. Debabov","doi":"10.1134/s0003683824604360","DOIUrl":"https://doi.org/10.1134/s0003683824604360","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The biosynthesis of butyric acid from glucose through inverted fatty acid β-oxidation by recombinant <i>Escherichia coli</i> strains was optimized. An increased yield of the target compound was achieved resulting from the plasmid expression of the <i>atoB</i>, <i>fadB,</i> and <i>fadE</i>/<i>fabI</i> genes in the core strain MG∆4 P<sub>L</sub>-<i>tesB</i> Δ<i>yciA</i> (MG1655 ∆<i>ackA-pta</i>, ∆<i>poxB</i>, ∆<i>ldhA</i>, ∆<i>adhE</i>, P<sub>L</sub>-SD<sub>ϕ10</sub>-<i>tesB</i>, ∆<i>yciA</i>). The positive effect of enforced ATP hydrolysis on microaerobic conversion of the carbohydrate substrate to the final product by the recombinants was demonstrated. Activation of the futile cycle of pyruvate–phosphoenolpyruvate–pyruvate, due to the increased expression of the <i>ppsA</i> gene, ensured a marked increase in glucose consumption by the recombinants and led to an increase in the molar yield of butyric acid up to 39.5%. When the components of the H<sup>+</sup>-ATP synthase complex were uncoupled resulting from the deletion of <i>atpFH</i> genes, the molar yield of butyric acid from glucose demonstrated by the strain forming butyryl-CoA by the action of enoyl-ACP reductase FabI reached 46%.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141782994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604451
D. N. Zuev, E. I. Cherkasova, K. V. Apryatina, S. D. Zaitsev, L. A. Smirnova
Abstract
Grafted copolymers of chitosan–vinylpyrrolidone, water-soluble at a pH of 6.8–7.5, were obtained. A technique has been developed for obtaining an aggregatively stable system of platinum nanoparticles in copolymer solutions with an average size of ~4 nm. The thermophysical and structural characteristics of the powdered composition of a platinum nanoparticle-copolymer are investigated. An in vitro comparison of the antitumor activity of solutions of the developed composition and cisplatin at the same platinum concentration was performed. It was found that with respect to the culture of HeLa Kyoto and A431 cancer cells, the composition is five and two times less effective than cisplatin, respectively. Along with this, the biocompatibility of the composition is 17 times higher than that of cisplatin, which allows its use at elevated concentrations and the development of an antitumor agent with platinum nanoparticles commensurate in effectiveness with cisplatin.
{"title":"Platinum Nanoparticles in Aqueous Solutions of a Chitosan–Vinylpyrrolidone Copolymer: Synthesis and Biological Activity","authors":"D. N. Zuev, E. I. Cherkasova, K. V. Apryatina, S. D. Zaitsev, L. A. Smirnova","doi":"10.1134/s0003683824604451","DOIUrl":"https://doi.org/10.1134/s0003683824604451","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>Grafted copolymers of chitosan–vinylpyrrolidone, water-soluble at a pH of 6.8–7.5, were obtained. A technique has been developed for obtaining an aggregatively stable system of platinum nanoparticles in copolymer solutions with an average size of ~4 nm. The thermophysical and structural characteristics of the powdered composition of a platinum nanoparticle-copolymer are investigated. An in vitro comparison of the antitumor activity of solutions of the developed composition and cisplatin at the same platinum concentration was performed. It was found that with respect to the culture of HeLa Kyoto and A431 cancer cells, the composition is five and two times less effective than cisplatin, respectively. Along with this, the biocompatibility of the composition is 17 times higher than that of cisplatin, which allows its use at elevated concentrations and the development of an antitumor agent with platinum nanoparticles commensurate in effectiveness with cisplatin.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141782996","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s000368382460444x
S. O. Rogozhkin, A. S. Gerasimov
Abstract
CRM197 (Cross Reacting Material 197) is an inactive form of the C. diphtheriae exotoxin used as a carrier protein for the development and production of conjugated polysaccharide vaccines and immunotherapeutic drugs. However, the development of these research areas is not possible without an efficient and cost-effective technology to produce CRM197 of the proper quality. In this study, we developed a highly efficient method to produce recombinant CRM197 as a fusion with SUMO protein, yielding more than three grams per liter in the form of inclusion bodies. We examined the significant effect of the type of expression vector, the heterologous gene expression conditions, and cultivation on its solubility. Using a combination of reduced cultivation temperature and the promoter of the gene encoding the heat shock protein CspA, we achieved an increase in the solubility level of SUMO-CRM197 of more than 30%, with an overall biosynthesis level of more than two grams per liter. Coexpression of the target gene with the DsbC disulfide isomerase gene allowed us to obtain the target protein completely in the soluble state with a yield of more than 1.4 grams per liter. The results obtained may become the basis for the development of a promising domestic technology for the production of CRM197.
{"title":"Bacterial Expression of Crm197: Investigation and Optimization of Gene Expression Factors for Effective Production in E. coli","authors":"S. O. Rogozhkin, A. S. Gerasimov","doi":"10.1134/s000368382460444x","DOIUrl":"https://doi.org/10.1134/s000368382460444x","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>CRM197 (<b>C</b>ross <b>R</b>eacting <b>M</b>aterial 197) is an inactive form of the <i>C. diphtheriae</i> exotoxin used as a carrier protein for the development and production of conjugated polysaccharide vaccines and immunotherapeutic drugs. However, the development of these research areas is not possible without an efficient and cost-effective technology to produce CRM197 of the proper quality. In this study, we developed a highly efficient method to produce recombinant CRM197 as a fusion with SUMO protein, yielding more than three grams per liter in the form of inclusion bodies. We examined the significant effect of the type of expression vector, the heterologous gene expression conditions, and cultivation on its solubility. Using a combination of reduced cultivation temperature and the promoter of the gene encoding the heat shock protein CspA, we achieved an increase in the solubility level of SUMO-CRM197 of more than 30%, with an overall biosynthesis level of more than two grams per liter. Coexpression of the target gene with the DsbC disulfide isomerase gene allowed us to obtain the target protein completely in the soluble state with a yield of more than 1.4 grams per liter. The results obtained may become the basis for the development of a promising domestic technology for the production of CRM197.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141783000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604426
L. E. Makarova, Yu. A. Markova, Yu. V. Zaytseva, A. A. Bychkova, I. V. Gorbenko, Yu. M. Konstantinov, I. A. Vasiliev, A. S. Morits, P. A. Bizikov
Abstract
Previously it was shown that endophytic bacteria had the ability to move out of the roots of pea plant seedlings (Pisum sativum L.) into the rhizosphere. In this study, six distinct bacterial strains were isolated from the root growth medium during the cultivation of seedlings in an aqueous medium. By analyzing the nucleotide sequence of 16S rRNA genes, the taxonomic position of these strains was established. Their morphological and cultural parameters were assessed, and the activity of hydrolytic enzymes (pectinase, cellulase, protease) and the IAA-producing capability were examined. It has been observed that the quantity of endophytic bacteria that appears on the root surface during the growth of pea seedlings significantly surpasses the quantity present in the root tissues. It is assumed that hydrolytic enzymes such as pectinase and cellulase are involved in the release of bacteria into the external environment, causing the destruction of carbohydrate structures in plant cell walls. The metabolic parameters established in the studied strains, and the significance of these endophytic bacteria for the host plant after their exit from the roots into the rhizosphere are under discussion.
{"title":"Phylogeny and Characterization of Endophytic Bacteria Isolated from the Rhizosphere of Pea Seedlings (Pisum Sativum L.)","authors":"L. E. Makarova, Yu. A. Markova, Yu. V. Zaytseva, A. A. Bychkova, I. V. Gorbenko, Yu. M. Konstantinov, I. A. Vasiliev, A. S. Morits, P. A. Bizikov","doi":"10.1134/s0003683824604426","DOIUrl":"https://doi.org/10.1134/s0003683824604426","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>Previously it was shown that endophytic bacteria had the ability to move out of the roots of pea plant seedlings (<i>Pisum sativum L</i>.) into the rhizosphere. In this study, six distinct bacterial strains were isolated from the root growth medium during the cultivation of seedlings in an aqueous medium. By analyzing the nucleotide sequence of 16S rRNA genes, the taxonomic position of these strains was established. Their morphological and cultural parameters were assessed, and the activity of hydrolytic enzymes (pectinase, cellulase, protease) and the IAA-producing capability were examined. It has been observed that the quantity of endophytic bacteria that appears on the root surface during the growth of pea seedlings significantly surpasses the quantity present in the root tissues. It is assumed that hydrolytic enzymes such as pectinase and cellulase are involved in the release of bacteria into the external environment, causing the destruction of carbohydrate structures in plant cell walls. The metabolic parameters established in the studied strains, and the significance of these endophytic bacteria for the host plant after their exit from the roots into the rhizosphere are under discussion.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141782999","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604359
A. N. Berlina, N. I. Smirnova, N. S. Komova, K. V. Serebrennikova, A. V. Zherdev, B. B. Dzantiev
Abstract
The influence of organic solvents such as methanol and acetonitrile on the results of butachlor immunoassay in samples of rice and rice products was studied. The schemes of enzyme immunoassay using: (a) native antiserum containing specific antibodies to butachlor and antispecies antibodies labeled with horseradish peroxidase, and (b) biotinylated specific antibodies and streptavidin labeled with peroxidase are considered. The close values of IC10 (0.77 and 0.68 ng/mL, respectively) and working range (2.6–165 and 2.4–192 ng/mL, respectively) were established for the two schemes, when analyzing in a mixture of phosphate buffer and methanol in a percentage ratio of 85 : 15, respectively. For the second scheme, the detection of butachlor in butachlor in rice-containing products is shown at 80-132% of the added analyte. A comparison of sample preparation methods allows us to recommend: (a) extraction of butachlor with hexane and re-dissolution of the dry residue in a buffer with 15% methanol or (b) preparation of a methanol extract followed by 6-fold dilution with a buffer solution.
{"title":"Influence of Organic Solvents on the Results of Immunoenzyme Determination of Herbicide Butachlor: Selection of Sample Preparation Modes","authors":"A. N. Berlina, N. I. Smirnova, N. S. Komova, K. V. Serebrennikova, A. V. Zherdev, B. B. Dzantiev","doi":"10.1134/s0003683824604359","DOIUrl":"https://doi.org/10.1134/s0003683824604359","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The influence of organic solvents such as methanol and acetonitrile on the results of butachlor immunoassay in samples of rice and rice products was studied. The schemes of enzyme immunoassay using: (a) native antiserum containing specific antibodies to butachlor and antispecies antibodies labeled with horseradish peroxidase, and (b) biotinylated specific antibodies and streptavidin labeled with peroxidase are considered. The close values of IC10 (0.77 and 0.68 ng/mL, respectively) and working range (2.6–165 and 2.4–192 ng/mL, respectively) were established for the two schemes, when analyzing in a mixture of phosphate buffer and methanol in a percentage ratio of 85 : 15, respectively. For the second scheme, the detection of butachlor in butachlor in rice-containing products is shown at 80-132% of the added analyte. A comparison of sample preparation methods allows us to recommend: (a) extraction of butachlor with hexane and re-dissolution of the dry residue in a buffer with 15% methanol or (b) preparation of a methanol extract followed by 6-fold dilution with a buffer solution.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141783002","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604335
I. V. Kravchenko, V. A. Furalyov, E. S. Pshennikova, E. V. Kostyleva, A. S. Sereda, E. I. Kurbatova, N. V. Tsurikova, A. N. Fedorov, V. O. Popov
Abstract
The effect of four enzyme preparations (EP), Bacillolysin, Agroprot, Protozyme, and Protozyme C (Russia), on the protein and peptide profiles of the protein isolate isolated from peas of the Focor variety, as well as on its smell and taste, was investigated in this work. It was shown that enzyme treatment can improve the odor characteristics of the isolate. Thus, it was possible to reduce significantly the severity of the bean and herbal smell. At the same time, enzyme treatment also improved the taste of the isolate: it was possible to reduce significantly the severity of disturbing flavors such as leguminous, astringent, bitter, and herbal. The results obtained allowed us to select EP (fungal acid aspartic protease) to improve the organoleptic parameters of pea protein isolates intended for the production of meat and dairy product analogs.
{"title":"The Effect of Various Domestically Produced Proteolytic Enzyme Preparations on the Organoleptic Properties of Pea Protein Isolates","authors":"I. V. Kravchenko, V. A. Furalyov, E. S. Pshennikova, E. V. Kostyleva, A. S. Sereda, E. I. Kurbatova, N. V. Tsurikova, A. N. Fedorov, V. O. Popov","doi":"10.1134/s0003683824604335","DOIUrl":"https://doi.org/10.1134/s0003683824604335","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Abstract</h3><p>The effect of four enzyme preparations (EP), Bacillolysin, Agroprot, Protozyme, and Protozyme C (Russia), on the protein and peptide profiles of the protein isolate isolated from peas of the Focor variety, as well as on its smell and taste, was investigated in this work. It was shown that enzyme treatment can improve the odor characteristics of the isolate. Thus, it was possible to reduce significantly the severity of the bean and herbal smell. At the same time, enzyme treatment also improved the taste of the isolate: it was possible to reduce significantly the severity of disturbing flavors such as leguminous, astringent, bitter, and herbal. The results obtained allowed us to select EP (fungal acid aspartic protease) to improve the organoleptic parameters of pea protein isolates intended for the production of meat and dairy product analogs.</p>","PeriodicalId":466,"journal":{"name":"Applied Biochemistry and Microbiology","volume":null,"pages":null},"PeriodicalIF":0.8,"publicationDate":"2024-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141782995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-07-27DOI: 10.1134/s0003683824604372
N. D. Murtazina, L. S. Sharapova, N. P. Yurina
Abstract
The chaperone system of the cell is the first line of defense in plants under stress. In the present work, the effect of heat stress on the levels of cytoplasmic HSP70 and chloroplast HSP70B chaperones in three Cucurbita species (C. maxima Duchesne, C. pepo L., and C. moschata, Duchesne) differing in their stress resistance was studied. A correlation between the levels of cytoplasmic HSP70 and chloroplast HSP70B chaperones and pumpkin species under heat stress was demonstrated. Under stress, a significant increase in the chaperone levels was observed in the cells of the C. maxima pumpkin, namely, the level of the cytoplasmic HSP70 increased by 3.6 times, and the level of chloroplast HSP70B increased by two times. Heat stress caused a 1.7-fold increase in the level of the cytoplasmic chaperone HSP70 in the cells of the C. pepo pumpkin, while no significant change in the level of the HSP70B protein was observed. However, heat stress led to a decrease in the levels of both HSP70 and HSP70B compared to untreated plants in the C. moschata pumpkin. The dynamics of changes in the levels of cytoplasmic and chloroplast chaperones under heat stress is similar. It should be noted that the constitutive levels of HSP70 and HSP70B under normal conditions are higher in C. moschata and C. pepo compared to C. maxima. The analysis of the obtained data revealed an interesting pattern: high constitutive levels of HSPs result in an insignificant induction of HSPs, and, vice versa, low constitutive levels of these proteins correlate with the high induction rate of these proteins after exposure to heat stress. These findings are important in understanding the mechanisms of plant stress resistance and may be useful for selection and development of highly resistant productive varieties of agriculturally important plants.
摘要 细胞伴侣系统是植物在胁迫下的第一道防线。本文研究了热胁迫对三种抗逆性不同的葫芦科植物（C. maxima Duchesne、C. pepo L. 和 C. moschata, Duchesne）细胞质 HSP70 和叶绿体 HSP70B 合子水平的影响。结果表明，在热胁迫下，细胞质 HSP70 和叶绿体 HSP70B 合子的水平与南瓜品种之间存在相关性。在胁迫条件下，观察到 C. maxima 南瓜细胞中的伴侣素水平显著增加，即细胞质 HSP70 水平增加了 3.6 倍，叶绿体 HSP70B 水平增加了 2 倍。热胁迫导致 C. pepo 南瓜细胞中细胞质伴侣蛋白 HSP70 的水平增加了 1.7 倍，而 HSP70B 蛋白的水平没有发生显著变化。然而，与未处理的植物相比，热胁迫导致 C. moschata 南瓜中 HSP70 和 HSP70B 的水平下降。在热胁迫下，细胞质和叶绿体伴侣蛋白水平的动态变化相似。值得注意的是，在正常条件下，C. moschata 和 C. pepo 中 HSP70 和 HSP70B 的组成水平要高于 C. maxima。对所获数据的分析揭示了一种有趣的模式：高组成水平的 HSPs 导致 HSPs 的诱导不明显，反之亦然，这些蛋白质的低组成水平与暴露于热胁迫后这些蛋白质的高诱导率相关。这些发现对于了解植物抗逆性的机制非常重要，可能有助于选育高抗逆性的重要农业植物品种。
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