State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika

Moscow, Russia

State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika

Moscow, Russia
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Borshchevskaya L.N.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Gordeeva T.L.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Sineoky S.P.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Applied Biochemistry and Microbiology | Year: 2016

The expression of L-lactate dehydrogenase genes ldh1 (Bos taurus), ldhA (Homo sapiens), ldhA (Rhizopus oryzae), ldh1 (Lactobacillus plantarum), and ldh1 (Lactobacillus pentosus) in the cells of yeast Schizosaccharomyces pombe VKPM U-3106 has been investigated. The catalytic characteristics of the enzymes encoded by these genes have been compared, and the intensity of lactic acid synthesis by the recombinant strains obtained has been evaluated. The enzymatic activity of L-lactate dehydrogenases from L. plantarum and L. pentosus was the highest (approximately 2 to 2.5 times higher than that of the mammalian enzymes), and these enzymes therefore appear to have the highest potential for the development of lactic-acid producing strains of yeast S. pombe. © 2016, Pleiades Publishing, Inc.


Kotova L.N.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Serebrennikov V.M.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Glazunov A.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Applied Biochemistry and Microbiology | Year: 2016

The influence of population heterogeneity and medium pH factors on the effect of α-acetolactate (AL) overproduction that we found in L. lactis bv. diacetylactis B2103/74 culture was studied. The nature of the enzymatic system responsible for this effect (which is able to considerably block lactate dehydrogenase (LDH) by intercepting up to 80% of the NADH in it) is unknown; we designated this system as the heminindependent electron transferring system (HEMIETS). It was demonstrated that manifestation of the effect was masked by the culture heterogeneity and was especially strong in approximately neutral pH values (6.1–6.5). The presence in the population of at least three types of dissociants differing in the level of HEMIETS activity, and consequently AL synthesis, was established: an active dissociant (AL accumulation up to 30 mM), an inactive dissociant that had lost the HEMIETS (AL no more than 3 mM), and a dissociant with intermediate activity (AL 15–17 mM). It was demonstrated that the HEMIETS activity at a particular pH value had primary importance in the emerging general picture of pyruvate metabolism at different pH values and, finally, in overproduction. Thus, no overproduction effect was observed at a pH of 7.0 due to the low HEMIETS activity, and the pyruvate metabolism passed according to homolactic type. However, the HEMIETS activity was so high at pH values of 5.3–6.5 that this allowed it to take the function of the main regulator of NAD+/NADH oxidation–reduction balance and to give a minor role to LDH. The LDH- and acetolactate synthase pathway productivity depended on the HEMIETS activity at a particular pH, while the productivity of pyruvate dehydrogenase pathway directly depended on the external pH. The latter was relatively high in approximately neutral area (pH 6.0–6.5) and was noticeably decreased in neutral (pH 7.0) and weakly acid (pH ≤ 6.0) zones. © 2016, Pleiades Publishing, Inc.


Yuzbashev T.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Bondarenko P.Y.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Sobolevskaya T.I.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Yuzbasheva E.Y.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | And 6 more authors.
Biotechnology and Bioengineering | Year: 2016

Bio-based succinic acid production can redirect industrial chemistry processes from using limited hydrocarbons to renewable carbohydrates. A fermentation process that does not require pH-titrating agents will be advantageous to the industry. Previously, a Yarrowia lipolytica strain that was defective for succinate dehydrogenase was constructed and was found to accumulate up to 17.5 g L−1 of succinic acid when grown on glycerol without buffering. Here, a derivative mutant was isolated that produced 40.5 g L−1 of succinic acid in 36 h with a yield of 0.32 g g−1 glycerol. A combination approach of induced mutagenesis and metabolic evolution allowed isolation of another derivative that could utilize glucose efficiently and accumulated 50.2 g L−1 succinic acid in 54 h with a yield of 0.43 g g−1. The parent strain of these isolated mutants was used for [1,6-13C2]glucose assimilation analysis. At least 35% glucose was estimated to be utilized through the pentose phosphate pathway, while ≥84% succinic acid was formed through the oxidative branch of the tricarboxylic acid cycle. Biotechnol. Bioeng. 2016;113: 2425–2432. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.


PubMed | RAS N. D. Zelinsky Institute of Organic Chemistry and State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Type: Journal Article | Journal: Biotechnology and bioengineering | Year: 2016

Bio-based succinic acid production can redirect industrial chemistry processes from using limited hydrocarbons to renewable carbohydrates. A fermentation process that does not require pH-titrating agents will be advantageous to the industry. Previously, a Yarrowia lipolytica strain that was defective for succinate dehydrogenase was constructed and was found to accumulate up to 17.5gL(-1) of succinic acid when grown on glycerol without buffering. Here, a derivative mutant was isolated that produced 40.5gL(-1) of succinic acid in 36h with a yield of 0.32gg(-1) glycerol. A combination approach of induced mutagenesis and metabolic evolution allowed isolation of another derivative that could utilize glucose efficiently and accumulated 50.2gL(-1) succinic acid in 54h with a yield of 0.43gg(-1) . The parent strain of these isolated mutants was used for [1,6-(13) C2 ]glucose assimilation analysis. At least 35% glucose was estimated to be utilized through the pentose phosphate pathway, while 84% succinic acid was formed through the oxidative branch of the tricarboxylic acid cycle. Biotechnol. Bioeng. 2016;113: 2425-2432. 2016 Wiley Periodicals, Inc.


Yuzbashev T.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Larina A.S.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Vybornaya T.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Yuzbasheva E.Y.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | And 2 more authors.
Fungal Biology | Year: 2015

The vast number of repetitive genomic elements was identified in the genome of Rhizopus oryzae. Such genomic repeats can be used as homologous regions for integration of plasmids. Here, we evaluated the use of two different repeats: the short (575bp) rptZ, widely distributed (about 34 copies per genome) and the long (2053 bp) rptH, less prevalent (about 15 copies). The plasmid carrying rptZ integrated, but did so through a 2256-bp region of homology to the pyrG locus, a unique genomic sequence. Thus, the length of rptZ was below the minimal requirements for homologous strand exchange in this fungus. In contrast, rptH was used efficiently for homologous integration. The plasmid bearing this repeat integrated in multicopy fashion, with up to 25 copies arranged in tandem. The latter vector, pPyrG-H, could be a valuable tool for integration at homologous sequences, for such purposes as high-level expression of proteins. © 2015 The British Mycological Society.


Mordkovich N.N.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Manuvera V.A.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Veiko V.P.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Debabov V.G.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Applied Biochemistry and Microbiology | Year: 2012

A DNA fragment containing a promoter-operator and structural parts of the uridine phosphorylase gene from Shewanella oneidensis MR-1 was cloned. Cross-heterological expression of the udp genes from Sh. oneidensis MR-1 and Escherichia coli under the control of authentic regulatory regions is shown. The UDP protein accumulates in an active form in the cytoplasmic fraction of cells. The recombinant UDP protein from Sh. oneidensis MR-1 obtained by heterological expression was isolated and characterized. E. coli udp gene promoter activity was observed during heterological expression in Sh. oneidensis MR-1 cells under both aerobic and anaerobic conditions. © 2012 Pleiades Publishing, Ltd.


Kurochkina V.B.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Sklyarenko A.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Satarova J.E.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Yarotsky S.V.,State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Bioprocess and Biosystems Engineering | Year: 2011

The article deals with experimental determination of ionization constants and solubility for the compounds (target products, initial β-lactams, acylating agents and by-products) involved in enzymatic synthesis of some therapeutically used aminopenicillins and aminocephalosporins, namely ampicillin, amoxicillin, cephalexin, cephadroxil, cephaloglycin, cefaclor, cefprozil, cefatrizine. Methodology of investigations and the evaluation of experimental data for the determination of ionization constants and solubility of the different type electrolytes are presented. Applications of the methods based on acid-base potentiometric titration and on determination of solubility-pH dependence of assayed substances are discussed. The original data on ionization constants and solubility of amoxicillin, cefprozil, cefatrizine, cephadroxil and initial β-lactams for production of cefaclor, cefprozil and cefatrizine, as well as solubility of by-product d-(-)-p-hydroxyphenylglycine are presented. Experimentally determined parameters and constants available in the literature for all abovementioned aminopenicillins and aminocephalosporins are collected. These data might be used for choice of the conditions of both processes: the enzymatic synthesis and the isolation of the product from reaction mixture. © 2011 Springer-Verlag.


PubMed | Moscow Institute of Physics and Technology and State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Type: | Journal: Microbiological research | Year: 2016

The antirestriction proteins ArdA ColIb-P9, Arn T4 and Ocr T7 specifically inhibit type I and type IV restriction enzymes and belong to the family of DNA-mimic proteins because their three-dimensional structure is similar to the double-helical B-form DNA. It is proposed that the DNA-mimic proteins are able to bind nucleoid protein H-NS and alleviate H-NS-silencing of the transcription of bacterial genes. Escherichia coli lux biosensors were constructed by inserting H-NS-dependent promoters into a vector, thereby placing each fragment upstream of the promoterless Photorhabdus luminescens luxCDABE operon. It was demonstrated that the DNA-mimic proteins ArdA, Arn and Ocr activate the transcription of H-NS-dependent promoters of the lux operon of marine luminescent bacteria (mesophilic Aliivibrio fischeri and psychrophilic Aliivibrio logei), and the dps gene from E. coli. It was also demonstrated that the ArdA antirestriction protein, the genes of which are located on transmissive plasmids ColIb-P9, R64, PK101, decreases levels of H-NS silencing of the PluxC promoter during conjugation in the recipient bacteria.


PubMed | State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Type: Journal Article | Journal: Bioprocess and biosystems engineering | Year: 2011

The article deals with experimental determination of ionization constants and solubility for the compounds (target products, initial -lactams, acylating agents and by-products) involved in enzymatic synthesis of some therapeutically used aminopenicillins and aminocephalosporins, namely ampicillin, amoxicillin, cephalexin, cephadroxil, cephaloglycin, cefaclor, cefprozil, cefatrizine. Methodology of investigations and the evaluation of experimental data for the determination of ionization constants and solubility of the different type electrolytes are presented. Applications of the methods based on acid-base potentiometric titration and on determination of solubility-pH dependence of assayed substances are discussed. The original data on ionization constants and solubility of amoxicillin, cefprozil, cefatrizine, cephadroxil and initial -lactams for production of cefaclor, cefprozil and cefatrizine, as well as solubility of by-product D-(-)-p-hydroxyphenylglycine are presented. Experimentally determined parameters and constants available in the literature for all abovementioned aminopenicillins and aminocephalosporins are collected. These data might be used for choice of the conditions of both processes: the enzymatic synthesis and the isolation of the product from reaction mixture.


PubMed | State Research Institute of Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Type: Journal Article | Journal: FEMS microbiology letters | Year: 2012

The mercury-resistance transposon Tn5053 inhibits restriction activity of the type I restriction-modification endonuclease EcoKI in Escherichia coli K12 cells. This is the first report of antirestriction activity of a non-conjugative transposon. The gene (ardD) coding for the antirestriction protein has been cloned. The ardD gene is located within the tniA gene, coding for transposase, on the complementary strand. The direction of transcription is opposite to transcription of the tniA gene.

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