A. N. Kalinina's research while affiliated with Kurchatov Institute and other places

Publications (28)

Article
The phyCg gene encoding a new phytase from C. gillenii was optimized, synthesized, cloned, and expressed in Pichia pastoris. Analysis of the amino acid sequence of the enzyme showed that it belongs to the histidine acid phosphatase family. The amino acid sequence of the PhyCg phytase has the highest homology (73.49%) with a phytase sequence from Ci...
Article
The isolation, heterologous gene expression, and characteristics of a new β-glucanase from Bacillus pumilus is described. The bgl1 gene from the Bacillus pumilus Bg57 VKPM В-13195 strain, which consists of 729 nucleotides, encodes a secreted endo-β-l,3(4)-D-glucanase (EC 3.2.1.6) containing 214 amino acids and 28 residues of the putative signal pep...
Article
In¹ order to increase the expression level of gene for recombinant phytase in Pichia pastoris, a synthetic sequence of the phyOp-mod gene, which encodes phytase from Obesumbacterium proteus, has been designed and optimized by its codon composition. The GC content was adjusted to 48.33%, and AT-rich segments were eliminated. The main characteristics...
Article
The expression of four genes of bacterial endo-β-l,3(4)-D-glucanases from B. pumilus Bg57 (VKPM В-13195), Paenibacillus polymyxa Bg23 (VKPM В-4056), Bacillus subtilis Bg11 (VKPM B-13178), and Bacillus amyloliquefaciens Bg76 (VKPM B-13184), which are members of the family of GH16 glycohydrolases, in the expression system of Pichia pastoris are compa...
Article
The heterologous expression, isolation, and characterization of a novel xylanase from Paenibacillus brasilensis are described. The xyl1 gene from the Paenibacillus brasilensis strain X1 VKPM B-13092, which consists of 639 nucleotides, encodes a secreted endo-1,4-β-xylanase (EC 3.2.1.8) containing 184 amino acids and 28 residues of the putative sign...
Article
Endo-1,4-β-xylanases from Bacillus pumilus VKPM В-7975, Paenibacillus polymyxa VKPM В-3015, Thermomyces lanuginosus VKPM F-224, and Schizophyllum commune VKPM F-433, which are members of a family of 11 glycohydrolases, have been obtained in the expression system of Pichia pastoris. Technologically valuable characteristics of the recombinant hydrola...
Article
Phytases are widely used to improve the properties of mixed fodder. Due to its high specific activity, PhyА-Cf phytase from Citrobacter freundii is of special interest for agrobiotechnology. In order to decrease the thermal sensitivity of the enzyme during fodder granulation at 60–80°C, the thermal stability of phytase was enhanced by site-directed...
Article
The co-expression of genes encoding β-glucanases from Paenibacillus jamilae and Rhizomucor miehei in the Komagataella kurtzmanii methylotrophic yeast system has been carried out. The recombinant strains that simultaneously produced both enzymes with a total activity of 1132 and 1464 units for 96 h of cultivation in flasks were obtained. The combina...
Article
A DNA sequence from Kosakonia sacchari that according to automated computer analysis is believed to correspond to a gene for histidine acid phytase has been selected from the GenBank database. The sequence was optimized for codon composition, synthesized, cloned and expressed in Pichia pastoris. Main characteristics of the purified recombinant enzy...
Article
The isolation, heterologous expression and characterization of a new thermostable β-glucanase from Paenibacillus jamilae is described. The bgl26 gene from the P. jamilae Bg1 VKPM B-13093 strain consisting of 714 nucleotides encodes endo-1,3-1,4-β-glucanase (EC 3.2.1.73) containing 213 amino acids and 24 residues of the putative signal peptide in N-...
Article
Effect of overexpression of the HAC1 genes from Pichia pastoris and Saccharomyces cerevisiae on the production of heterologous enzymes, Escherichia coli phytase and Paenibacillus brasilensis xylanase, in P. pastoris cells has been studied. Codon composition of the phytase and xylanase encoding genes was optimized, and the genes were expressed in P....
Article
A method is proposed for the rapid identification of the taxa of five industrially valuable yeast groups: Pichia pastoris, Yarrowia lipolytica, Schizosaccharomyces pombe, Saccharomyces cerevisiae, and Kluyveromyces lactis. The specific primers selected based on nucleotide sequences of genes encoding rRNA were used in PCR. The method was effective i...
Article
A stepwise screening of bacterial xylanase producers isolated from forest soil in Moscow oblast has been performed, and the taxa of the microorganisms have been identified based on an analysis of the 16S RNA gene sequences. An approach to the selection of the most active producers is suggested, and this selection has been carried out, resulting in...
Article
Comparative analysis of the efficiency of the expression of the Escherichia coli, Obessumbacterium proteus, and Citrobacter freundii phytase genes in Pichia pastoris methylotrophic yeast has been carried out. A simple model with a genetic construct based on the pPICά vector and the plate test with a modified diagnostic medium containing methanol (i...
Article
Описана изоляция, гетерологическая экспрессия и характеристика новой ксиланазы из Paenibacillus brasilensis. Ген xyl1 из штамма P. brasilensis X1 ВКПМ В-13092, состоящий из 639 нуклеотидов, кодирует эндо-1,4-β-ксиланазу (EC 3.2.1.8) из 184 аминокислот и 28 остатков предполагаемого сигнального пептида в N-концевой области. Нуклеотидная последователь...
Article
In order to increase the level of expression of recombinant phytases in the Pichia pastoris yeast, a synthetic sequence of the phyOp-mod gene that encodes phytase from Obesumbacterium proteus has been developed and optimized by codon composition. The GC content was adjusted to 48.33, and the AT-rich segments were eliminated. The main characteristic...
Article
The expression of the genes for endo-1,4-β-xylanase from Bacillus pumilus VKPM â-7975, Paenibacillus polymyxa VKPM â-3015, Thermomyces lanuginosus VKPM F-224, and Schizophyllum commune VKPM F-433, members of the family of 11 glycohydrolases, in the expression system of Pichia pastoris has been assessed. Such technologically valuable characteristics...
Article
Фитазы широко используются для улучшения питательных свойств комбикормов. Благодаря высокой удельной активности фитаза PhyА-Cf из Citrobacter freundii представляет особый интерес для агробиотехнологии. Для снижения инактивации фермента во время гранулирования корма при 60-80 °С была увеличена термостабильность фитазы PhyА-Cf методом сайт-направленн...
Article
The isolation, heterological gene expression and characteristics of a new β-glucanase from Bacillus pumilus has been described for the first time. A gene bgl1 from the Bacillus pumilus Bg57 VKPM В-13195 consisting of 729 nucleotides encodes a secreted endo- β-l,3(4)-d-glucanase ( EC 3.2.1.6) that contains 214 amino acids and 28 residues of the puta...
Article
В экспрессионной системе Pichia pastoris проведено сравнение четырех бактериальных эндо-β- l,3(4)-D-глюканаз из Bacillus pumilus Bg57 ВКПМ В-13195, Paenibacillus polymyxa Bg23 ВКПМ В-4056, Bacillus subtilis Bg11 ВКПМ B-13178 и B. amyloliquefaciens Bg76 ВКПМ B-13184, относящихся к семейству GH16 гликозилгидролаз. Были исследованы такие технологическ...
Article
A stepwise screening of bacterial producers of xylanases isolated from the forest soil in Moscow oblast, and also the taxonomic identification of the microorganisms based on the analysis of the 16S RNA gene sequences have been performed. An approach to the selection of the most active producers was suggested, and this selection was carried out resu...
Article
A method for a rapid taxonomy identification of five industrially valuable yeast groups, Pichia pastoris, Yarrowia lipolytica, Schizosaccharomyces pombe, Saccharomyces cerevisiae and Kluyveromyces lactis, has been suggested. The specific primers that were selected on the basis of nucleotide sequences of genes encoding rRNA were used in PCR. The met...
Article
A comparative analysis of the efficiency of the Escherichia coli, Obessumbacterium proteus and Citrobacter freundii phytase gene expression in a Pichia pastoris methylotrophic yeast has been carried out. The level of the expression was assessed using a suggested for the first time simple model using a genetic construct based on the pPICά vector and...
Article
Full-text available
An efficient and inexpensive spectrophotometric method has been developed for the determination of lactic acid in the individual state and in food and biological and cultural liquids. The method is based on the spectrophotometric determination of the colored product of the reaction of lactate ions with iron(III) chloride at 390 nm. The optimum cond...
Article
A method for the taxonomic identification of seven closely related bacterial species of the Bacillus subtilis group (B. subtilis, B. amyloliquefaciens, B. licheniformis, B. vallismortis, B. atrophaeus, B. sonorensis, and B. mojavensis) using specific primers selected on the basis of the gyrA gene sequences was developed. The effectiveness of this m...

Citations

... Currently, the production of phytase by engineering strains has been achieved via high-density fermentation of recombinant P. pastoris and the specific activity with sodium phytate as a substrate is 1452 U/mg [1]. However, during traditional batch fermentation, continuous production is difficult to achieve because of frequent feeding and drawing [2,3]. ...
... Similar activation effects by Mn 2+ ,Co 2+ , Zn 2+ , Ca 2+ , Mg 2+ of 10 mM has been described for xylanase from A. fumigatus JCM 10,253 [61]. The activation effect of Co 2+ or Mn 2+ on GH 10 xylanase Xyn27, AfXylA10 and 11 xylanase Xyl1 with reported [62][63][64], while they inhibited Caldicoprobacter algeriensis XynBCA [4]. It is pointed out that the effect of metal ions on enzyme activity is closely related to xylanase structure. ...
... Recently, several studies on soil proteomes have discovered new microorganisms that produce new glucanases. However, most of these studies are still restricted to the chemical and molecular identification of enzymes (Zhou et al. 2016;Angelov et al. 2017;Borshchevskaya et al. 2019;Edison and Pradeep 2020). However, dos Gonçalves et al. (2020) identified a β-glucanase in mangrove soils, which is thermostable and active at basic pH. ...
... The phytase activity of the selected transformants was determined on a diagnostic medium with CaCl 2 , according to the method Gordeeva et al. 2018. The transformants were grown at 30 • C for 24 h. ...
... To this end, Bedford (2000) explored the use of genetic engineering to improve exogenous enzyme thermal stability and adaptation to the high temperatures of the process of feed preparation. Gordeeva et al. (2019) increased the thermal stability of enzymes used in feedstuffs by sitedirected saturation mutagenesis. Genetic engineering has been widely used to improve enzyme properties to promote their better application. ...
... It exhibits maximum activity at pH 4.9, thermal stability up to 60 • C (at pH 3.0) and a maximum activity of~340 FYT/mg. There are reports about production of this enzyme in Pichia pastoris including its artificially modified derivative with enhanced thermal stability [22][23][24]. ...
... The expression vector pPIC9 [12], which contains the inducible AOX1 promoter, the S. cervisiae factor signal sequence, and the selective marker HIS4 was used. The phyS3-mod gene with an optimized composition was synthesized at the Kurchatov Institute NRC of GOSNIIGENETIKA with the method in [13]. ...
... The fermented broth further had undergone downstream processing to separate the produced lactic acid from fermentation broth media as suggested by [24]. The microbe produced lactic acids were estimated through the spectrophotometric method [25] and the Acidity test [26]. Two Best LA Producing bacteria were identified from the fermentation experiment. ...
... The number of copies per genome of this ribosomal RNA operon varies between and within bacterial species, ranging from 6 to 15 for Bacillus sp. The developed BSG method, generating PCR amplicons with a size range from 137 to 165 bp, is the first realtime PCR method that was designed to specifically target the B. subtilis group in order to control the GMM contaminations in microbial fermentation products [28,30,32,33,39,40] (Tables 1 and S1). ...