State Research Institute for Genetics and Selection of Industrial Microorganisms

Moscow, Russia

State Research Institute for Genetics and Selection of Industrial Microorganisms

Moscow, Russia

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PubMed | Shumakov Research Institute of Transplantation and Artificial Organs, Moscow State University and State Research Institute for Genetics and Selection of Industrial Microorganisms
Type: | Journal: Doklady. Biochemistry and biophysics | Year: 2016

The study of the stimulating effect of the microgels (MGs) based on recombinant 1F9 spidroin on the regeneration of the deep skin wound in mice was carried out. The use of spidroin MGs was shown to increase significantly the quality of healing compared to the control. The introduction of the MG in the wound edges led to recovery of all the structural elements of the skin: the epidermis, the dermis, including vascular and nervous network, in the periphery of the wound underlying muscles, and skin appendages (sebaceous and sweat glands and hair follicles) was revealed.


PubMed | okhin Russian Cancer Research Center and State Research Institute for Genetics and Selection of Industrial Microorganisms
Type: Journal Article | Journal: Molecular biotechnology | Year: 2016

The modified asparaginase Was79 was derived from the recombinant wild-type L-asparaginase of Wolinella succinogenes. The Was79 contains the amino acid substitutions V23Q and K24T responsible for the resistance to trypsinolysis and the N-terminal heparin-binding peptide KRKKKGKGLGKKR responsible for the binding to heparin and tumor K562 cells in vitro. When tested on a mouse model of Fischer lymphadenosis L5178Y, therapeutic efficacy of Was79 was significantly higher than that of reference enzymes at all single therapeutic doses used (125-8000IU/kg). At Was79 single doses of 500-8000IU/kg, the complete remission rate of 100% was observed. The Was79 variant can be expressed intracellularly in E. coli as a less immunogenic formyl-methionine-free form at high per cell production levels.


Debabov V.G.,State Research Institute for Genetics and Selection of Industrial Microorganisms
Applied Biochemistry and Microbiology | Year: 2010

Deep desulfurization of oil and its fractions is currently performed by hydration at high temperature and hydrogen pressure, which makes the process rather expensive. Searches for alternative modes for desulfurization, among which is biodesulfurization, are intensely in progress. In this review, the following subjects are discussed: microorganisms capable of desulfurizing petroleum products, mechanisms of their activity, achievements in the field of process development, and disadvantages of the method. The existing level of knowledge is insufficient for immediate implementation of an industrial biotechnological process for sulfur elimination from oil and motor fuel and it can only be regarded as a medium-term (10-15 years) prospect. © 2010 Pleiades Publishing, Ltd.


Seregina T.A.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Shakulov R.S.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Debabov V.G.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Mironov A.S.,State Research Institute for Genetics and Selection of Industrial Microorganisms
Applied Biochemistry and Microbiology | Year: 2010

Multistage construction of an E. coli strain containing no foreign genes which is capable of producing butyrate has been carried out. At the first stage, deletions in gene fadR encoding a protein repressor of an operon for fatty acid degradation and gene aceF responsible for the synthesis of pyruvate dehydrogenase were introduced in the strain MG1655 genome. Then, a mutant obtained from the above strain by induced mutagenesis and capable of growth on ethanol as a sole carbon source under aerobic conditions was selected. It was shown that growth of the mutant on ethanol is provided by two mutations. One of them (a substitution: 257G → A) is located in the regulatory region of gene adhE that controls the synthesis of alcohol-dehydrogenase; the other, containing a substitution Glu568 → Lys, affects the structural portion of the gene. As a result of the consequent mutagenesis of the obtained strain and selection on indicating media, variants capable of growing on butyrate and butanol as sole carbon sources and putatively bearing mutations in gene atoC (encoding transcriptional activator of atoDAB operon) were selected. At the last stage of the work, gene atoB, encoding the synthesis of the thiolase II enzyme, was placed under the control of a constitutive promoter Ptet, and the functional allele of gene aceF was introduced. The resulting E. coli strain (ΔfadR, adhE, atoC, Ptet-atoB) accumulates 800 mg/l of butyrate upon growth on glucose-containing medium under semi-anaerobic (oxygen limited) conditions. Introduction of an additional deletion in gene ldhA encoding lactate dehydrogenase in the strain genome leads to a further growth of a butyrate production up to 1.3 g/l. © 2010 Pleiades Publishing, Ltd.


Zubasheva M.V.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Ganushkina L.A.,Moscow Medical Academy | Smirnova T.A.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Azizbekyan R.R.,State Research Institute for Genetics and Selection of Industrial Microorganisms
Applied Biochemistry and Microbiology | Year: 2010

The optimum conditions for growth, sporulation, and crystal-formation in four isolated crystal-forming strains of Bacillus laterosporus were determined. It was shown that culture broth and pellets of bacterial culture liquid possess larvicidal activity against larvae of mosquitoes A. stephensi and A. aegypti. The protein nature of crystal was shown. Crystals are monocomponent containing a protein with MM of 68 or 130 kDa. Purified protein crystals demonstrated larvicidal activity. Specific larvicidal activity of crystals of various strains essentially differed. High larvicidal activity of B. laterosporus strains allows for them to be recommended as producers of antimosquito biological preparations. © 2010 Pleiades Publishing, Ltd.


Kotova V.Y.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Manukhov I.V.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Zavilgelskii G.B.,State Research Institute for Genetics and Selection of Industrial Microorganisms
Applied Biochemistry and Microbiology | Year: 2010

We have constructed hybrid plasmids pColD and pIbpA in which transcription of reporter genes luxCDABE from Photohabdus luminescens occurred from induced promoters: SOS-promoter of gene cdaA (pColD-CA23) and heat shock gene ibpA (E. coli MG1655 genome), respectively. Main parameters were measured: threshold concentration of an inducer and maximal response and minimal time of response for E. coli lux-biosensors bearing hybrid plasmids pColD and pRecA (SOS-response); pIbpA and pGrpe (heat shock) and pKatG and pSoxS (oxidative stress). Mitomicyn C, cis-diaminedichloroplatinum (SOS-response), ethanol and pentachlorophenol (heat shock), and hydrogen peroxide and methyl viologen (oxidative stress) were used as luminescence inducers. It was shown that the SOS-biosensor pColD is superior in main characteristics to the earlier constructed biosensor pRecA, and the heat biosensor pIbpA is more efficient than the earlier constructed pGrpE. Lux-biosensors (pKatG) and (pSoxS) are high sensitive to hydrogen peroxide and methyl viologen, respectively. © 2010 Pleiades Publishing, Ltd.


Kulakova O.G.,Russian National Research Medical University | Tsareva E.Y.,Russian National Research Medical University | Lvovs D.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Favorov A.V.,Johns Hopkins University | And 2 more authors.
Pharmacogenomics | Year: 2014

Various diseases require the selection of preferable treatment out of available alternatives. Multiple sclerosis (MS), an autoimmune inflammatory/neurodegenerative disease of the CNS, requires long-term medication with either specific disease-modifying therapy (DMT) - IFN-β or glatiramer acetate (GA) - which remain the only first-line DMTs in all countries. A significant share of MS patients are resistant to treatment with one or the other DMT; therefore, the earliest choice of preferable DMT is of particular importance. A number of conventional pharmacogenetic studies performed up to the present day have identified the treatment-sensitive genetic biomarkers that might be specific for the particular drug; however, the suitable biomarkers for selection of one or another first-line DMT are remained to be found. Comparative pharmacogenetic analysis may allow the identification of the discriminative genetic biomarkers, which may be more informative for an a priori DMT choice than those found in conventional pharmacogenetic studies. The search for discriminative markers of preferable first-line DMT, which differ in carriage between IFN-β responders and GA responders as well as between IFN-β nonresponders and GA nonresponders, has been performed in 253 IFN-β-treated MS patients and 285 GA-treated MS patients. A bioinformatics algorithm for identification of composite biomarkers (allelic sets) was applied on a unified set of immune-response genes, which are relevant for IFN-β and/or GA modes of action, and identical clinical criteria of treatment response. We found the range of discriminative markers, which include polymorphic variants of CCR5, IFNAR1, TGFB1, DRB1 or CTLA4 genes, in different combinations. Every allelic set includes the CCR5 genetic variant, which probably suggests its crucial role in the modulation of the DMT response. Special attention should be given to the (CCR5 d+ IFNAR1 G) discriminative combination, which clearly points towards IFN-β treatment choice for carriers of this combination. As a whole the comparative approach provides an option for the identification of prognostic composite biomarkers for a preferable medication among available alternatives. © 2014 Future Medicine Ltd.


Cheperegin S.E.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Efremov B.D.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Kozlov D.G.,State Research Institute for Genetics and Selection of Industrial Microorganisms
Protein Expression and Purification | Year: 2010

In vivo and in vitro experiments showed that human serum albumin (HSA) co-precipitated with components of the commonly used yeast peptone dextrose (YPD) growth medium in aqueous solutions at pH <5. Yeast extract was found to be the primary component of YPD responsible for HSA precipitation. Among yeast extract constituents, RNAs are likely to be most important for HSA precipitation. HSA precipitation at pH <5 was reversible, so that HSA was easily re-solubilized by increasing pH above 6 with completely retained immunoreactivity. The co-precipitation and re-solubilization of HSA were solely pH-dependent and occurred almost instantly at room temperature. Practical aspects of the observed HSA co-precipitation are discussed. © 2010 Elsevier Inc. All rights reserved.


Moisenovich M.M.,Moscow State University | Pustovalova O.L.,Moscow State University | Yu Arhipova A.,Moscow State University | Vasiljeva T.V.,Moscow State University | And 6 more authors.
Journal of Biomedical Materials Research - Part A | Year: 2011

The goal of this study was to generate porous scaffolds from the genetically engineered protein, an analogue of Nephila clavipes spidroin 1 (rS1/9) and to assess the properties of new rS1/9 scaffolds essential for bioengineering. The salt leaching technique was used to make the rS1/9 scaffolds of interconnected macroporous structure with spontaneously formed micropores. The tensile strength of scaffolds was 18 ± 5 N/cm2. Scaffolds were relatively stable in a phosphate buffer but degraded in oxidizing environment after 11 weeks of incubation. Applicability of the recombinant spidroin 1 as a substrate for cell culture was demonstrated by successful 3T3 cells growth on the surface of rS1/9 films (270 ± 20 cells/mm2 vs. 97 ± 8 cells/mm2 on the glass surface, p < 0.01). The 3T3 fibroblasts readily proliferated within the rS1/9 scaffold (from initially plated 19 ± 2 cells/mm3 to 3800 ± 304 cells/mm 3 after 2 weeks). By this time, cells were uniformly distributed between the surface and deeper layers (27% ± 8% and 33% ± 4%, respectively; p > 0.05), whereas the initial distribution was 58% ± 7% and 11% ± 8%, respectively; p < 0.05). The rS1/9 scaffolds implanted subcutaneously into Balb/c mice were well tolerated. Over a 2-month period, the scaffolds promoted an ingrowth of de novo formed vascularized connective tissue elements and nerve fibers. Thus, scaffolds made of the novel recombinant spidroin 1 analogue are potentially applicable in tissue engineering. © 2010 Wiley Periodicals, Inc.


Lavrov K.V.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Larikova G.A.,State Research Institute for Genetics and Selection of Industrial Microorganisms | Yanenko A.S.,State Research Institute for Genetics and Selection of Industrial Microorganisms
Applied Biochemistry and Microbiology | Year: 2013

The enzymatic synthesis of N-substituted acrylamides (N-isopropyl acrylamide and N, N-dimethylaminopropyl acrylamide) was demonstrated for the first time. The Rhodococcus erythropolis 37 strain, exhibiting acylamidase activity, was used as a source of enzyme, and water-dissolved acrylamide and isopropylamine/dimethylaminopropylamine served as substrates. The optimum conditions for the synthesis of acrylamide N-substitutes were determined using N-isopropyl acrylamide. The yield of the product was maximum at pH 9.5-10.5, substrate (acrylamide/isopropylamine) ratio within the range from 1.3: 1 to 2: 1, and absolute substrate concentrations of 8.0 (acrylamide) and 4.0% (isopropylamine). These conditions allowed for the synthesis of 22 g/L of N-isopropyl acrylamide. © 2013 Pleiades Publishing, Inc.

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