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Search results for: IMMOBILIZOWANE ENZYMY
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Disruption of homocitrate synthase genes in Candida albicans affects growth but not virulence
PublicationDwa geny, LYS21 i LYS22, kodujące izoformy syntazy homocytrynianowej, enzymu katalizującego pierwszy wyjątkowy etap w szlaku biosyntezy lizyny, zostały rozbite z komórkach Candida albicans z wykorzystaniem strategii ''SAT flipper''. Podwójny mutant lys21delta/lys22delta nie wykazywał aktywności syntazy homocytrynianowej, natomiast charakteryzował się auksotrofią wzrostową wobec lizyny w podłożach minimalnych, co można było odwrócić...
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Construction of glucosamine-6-phosphate synthase with an internal hexahistydyl fragment
PublicationL-Glutamine: D-fructose-6-phosphate aminotransferase (EC 2.6.1.16), known also as glucosamine-6-phosphate (GlcN-6-P) syntase, catalyzes the first committed step in the amino sugar biosynthetic pathway in prokaryotic and eukaryotic organisms. The enzyme carries out a complex reaction involving ammonia transfer and sugar phosphate isomerization. GlcN-6-P syntase is an important point of metabolic control of amino sugar biosynthesis...
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Unieruchamianie rekombinowanej beta-galaktozydazy w reakcjach katalizowanych transglutaminazą
PublicationTermostabilną beta-galaktozydazę wyizolowaną z Escherichia coli zawierającej gen enzymu z Pyrococcus woesei, sieciowano transgluta-minazą na żelu krzemionkowym. Otrzymany preparat wykazuje w 70 st.C aktywność hydrolizy ONPG wynoszącą 11,573 U/g nośnika. Optymalne pH i temperatura działania enzymu wynosi odpowiednio 5,5 oraz 95 st.C. Preparat unieruchomionego enzymu zachowuje około 70 % początkowej aktywności po 1 godzinie inkubacji...
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Structural features responsible for control of the eukaryotic GlcN-6-P synthase activity
PublicationGlucosamine-6-phosphate (GlcN-6-P) synthase is responsible for catalysis of the first and practically irreversible step in hexosamine metabolism. As UDP-GlcNAc, the final product of the hexosamine path, constitutes one of the essential substrates for assembly of bacterial and fungal cell walls, the enzyme is an interesting target for antimicrobial therapy.The structure of E. coli enzyme, known since 2001, forms a dimer of two identical...
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The psychrotrophic yeast Sporobolomyces roseus LOCK 1119 as a source of a highly active aspartic protease for the in vitro production of antioxidant peptides
PublicationA psychrotrophic yeast strain producing a cold-adapted protease at low temperature was classified as Sporobolomyces roseus. In standard YPG medium, S. roseus LOCK 1119 synthesized an extracellular protease with an activity of approximately 560 U/L. Optimization of medium composition and process temperature considerably enhanced enzyme biosynthesis; an approximate 70% increase in activity (2060 U/L). The native enzyme was purified...
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Beta-D-galaktozydazy - źródła, właściwości i zastosowanie
PublicationArtykuł przeglądowy opisujący mechanizm działania enzymu beta-D-galaktozydazy, właściwości beta-D-galaktozydaz pochodzących z różnych mikroorganizmów, zarówno mezofilnych jak i psychrofilnych oraz termofilnych, a także możliwości zastosowania enzymu w przemysłowych procesach hydrolizy laktozy w mleku, serwatce oraz w reakcji syntezy galaktooligosacharydów.
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Molecular modelling of transition state analogue inhibitors of glucosamine-6-P synthase and glucose-6-P isomerase.
PublicationStruktura kompleksu 2-amino-2-deoksy-D-glucitolo-6-P (ADGP) z centrum aktywnym syntazy GlcN-6-P z E. coli została wykorzystAna jako punkt wyjścia do modelowania molekularnego analogów ADGP. Używając programu GROMOS96 wygenerowano konformacje analogów o najniższych energiach wewnętrznych, które następnie ''dokowano'' w centrum aktywnym enzymu. Dokonano syntezy wybranych związków i określono parametry kinetyczne i termodynamiczne...
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A new cold-adapted beta-D-galactosidase from the Antarctic Arthrobacter sp. 32c - gene cloning, overexpression, purification and properties
PublicationThe development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production. In this article, we present a new β-D-galactosidase as a candidate to be applied in the above mentioned biotechnological...
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Characterization of recombinant homocitrate synthase from Candida albicans
PublicationLYS21 and LYS22 genes from Candida albicans encoding isoforms of homocitrate synthase (HCS), an enzyme catalyzing the first committed step in the L-lysine biosynthetic pathway, were cloned and expressed as NoligoHistagged fusion proteins in Escherichia coli. The purified gene products revealed HCS activity, i.e. catalyzed the condensation of α-ketoglutarate with acetyl-coenzyme A to yield homocitrate. The recombinant enzymes were purified...
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Heterogeneity of quaternary structure of glucosamine-6-phosphate deaminase from Giardia lamblia
PublicationThe oligoHis-tagged versions of glucosamine-6- phosphate deaminase from Giardia lamblia (GlmNagB-HisN, GlmNagB-HisC) were constructed and purified to hear homogeneity, and their kinetic and structural properties were compared to those of the wild-type enzyme (GlmNagB). Introduction of the oligoHis tag at the GlmNagB C-terminus resulted in almost complete loss of the catalytic activity, while the catalytic properties of GlmNagB-HisN...
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A new cold-active β-galactosidase from Arthrobacter sp. S3* - gene cloning, overexpression, purification and properties
PublicationA psychrotrophic bacterium producing a cold-active β-galactosidase was isolated from Spitsbergen soil and classified as Arthrobacter sp. S3*. The gene encoding β-galactosidase was isolated from the genomic DNA library, sequenced, cloned, expressed in Escherichia coli, purified by ion exchange chromatography and characterized. The Arthrobaster sp. S3* β-galactosidase is a homotrimeric enzyme composed of 74,4 kDa subunits. It is...
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Differential ability of cytostatics from anthraquinone group to generatefree radicals in three enzymatic systems: NADH dehydrogenase, NADPH cytochrome P450 reductase, and xantine oxidase.
PublicationZbadano szereg cytostatyków z grupy antrachinonu pod względem ich zdolności do pośredniczenia w przeniesieniu jednoelektronowym i generowaniu wolnych rodników tlenowych pod wpływem trzech oksydoreduktaz. Zdefiniowano czynniki strukturalne związków odpowiedzialne za ten proces katalizowany przez poszczególne enzymy.
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Extracellular proteolytic activity of Deinococcus geothermalis
PublicationProduction of extracellular protease by extremophilic bacteria Deinococcus geothermalis cultivated in liquid media containing 0.1% (w/v) of peptone K, 0.1% yeast extract and 0.2% marine salt reached a maximum in 14 h of the cell growth at 45°C and pH 8.0. The enzyme was purified by a two-step procedure using fractionation by a graded ammonium sulphate precipitation technique and gel filtration on Sephadex G-100 column. Protease...
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Docking simulations, Molecular properties and ADMET studies of novel Chromane6,7diol analogues as potential inhibitors of Mushroom tyrosinase
PublicationResearch on inhibition of tyrosinase enzyme has attained significant value, because tyrosinase inhibitors have potential applications in medicine, cosmetics (as whitening agents) and in agriculture (as bioinsecticides). Determination and elucidation of new tyrosinase inhibitors are not only beneficial for medical purposes, but their promising applications in improving food quality and nutritional...
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Fragmentomic analysis of biopeptides in silico released from milk proteins
PublicationThe fragmentomic-assisted method was employed to predict the biological potential of peptides derived from milk proteins hydrolyzed by papain and bromelain. Firstly, protein sequences were acquired from the BIOPEP-UWM database and then hydrolyzed by the above enzymes using a BIOPEP-UWM tool called “Enzyme(s) action”. The released peptides were defined as parent peptides and further analyzed for the presence of shorter peptidic...
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Acridinone antitumor agents, C-1305 and C-1299 are metabolized rather by flavin-containing monooxygenases and UDP-glucuronosyltransferases than cytochromes P450
PublicationStwierdzono, że to monooksygenazy flawinowe, a nie enzymy cytochromu P450, są głównymi enzymami frakcji mikrosomalnej, odpowiedzialnymi za detoksyfikacyjny metabolizm dwóch badanych pochodnych triazoloakrydonu. Dodatkowo C-1305 ulega sprzęganiu w czasie drugiej fazy metabolizmu z kwasem glukuronowym do postaci O-glukuronidu
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New source of the thermostable alpha-glucosidase suitable for single stepstarch processing
PublicationZbadana wewnątrzkomórkowa termostabilna alfa-glukozydaza z Thermus thermo-philus przejawia największą aktywność w 85 C przy pH 6.2. Aktywność enzymu po 5 h inkubacji w 75 C (pH 6.2) prawie nie ulega zmianie a okres połowicznej inaktywacji w 85 C wynosi 2 h. Właściwości te wskazują na przydatność wymienionego enzymu w jednoetapowej hydrolizie skrobi.
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Regulacja topologii DNA przez topoizomerazę typu II, czyli jak struktura DNA wpływa na funkcjonowanie genów
PublicationW wielu ważnych procesach jakie zachodzą w komórce zmienia się topologia DNA i struktura chromatyny. Enzym, topoizomeraza typu II, jest jednym z ważnych czynników, który reguluje zmiany topologiczne i strukturalne w jądrze, przez co ma bezpośredni lub pośredni wpływ na takie procesy jak uporządkowanie struktury chromatyny poprzez wiązanie pętli chromatyny do szkieletu jądrowego, transkrypcję i replikację DNA, reperację uszkodzeń...
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Characterization of glucoamylase immobilized on magnetic nanoparticles
PublicationMagnetic support was prepared by precipitation from an alkaline solution of divalent and trivalent iron ions and subsequently was modified with 3-aminopropyltriethoxysilane. FTIR analysis showed existence of a new Si-O-Fe bond in obtained particles. Scanning electronic microscopy images shows that the nanoparticles of all samples have particle size below 30 nm. Glucoamylase AMG 300L was immobilized onto the modified magnetic support...
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Systemy ekspresyjne białek cytochromu P450 w badaniach in vitro metabolizmu leków = Expression systems of cytochrome P450 proteins in studies of drug metabolism in vitro
PublicationBiałka cytochromu P450 to najważniejsze enzymy biorące udział w metabolizmie większości stosowanych w klinice leków, odpowiedzialne za ich aktywację lub detoksykację. Niektóre z dróg metabolizmu leku mogą być jednak odpowiedzialne za jego podwyższoną toksyczność. Nowe systemy ekspresyjne białek cytochromu P450 w komórkach ssaków, w tym człowieka, projektowane są w celu poznania roli metabolizmu w mechanizmie działania potencjalnych,...
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Synthesis of Trehalose by the Erythritol-Producing Yeast Yarrowia lipolytica Co-Displaying Maltooligosyltrehalose Synthase and Maltooligosyltrehalose Trehalohydrolase
PublicationIndustrial trehalose production faces economic challenges with costly enzyme preparations, prompting the exploration of eco-friendly alternatives. Here, we established a coupled functional sugar production line leveraging erythritolproducing cells as an innovative enzyme preparation for trehalose synthesis. The erythritol-producing Yarrowia lipolytica was modified to express a fusion protein consisting of maltooligosyltrehalose synthase...
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The extended version of restriction analysis approach for the examination of the ability of low-molecular-weight compounds to modify DNA in a cell-free system
PublicationOne of the primary requirements in toxicology is the assessment of ability of chemicals to induce DNA covalent modification. There are several well-established methods used for this purpose such as 32P-Postlabeling or HPLC-MS. However, all of these approaches have difficult to overcome limitations, which prevents their use in genotoxin screening. Here, we describe the simple protocol exploiting specificity of restriction enzymes...
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Fluorescent analogs of trypsin inhibitor SFTI-1 isolated from sunflower seeds-synthesis and applications
PublicationThis article describes the synthesis and enzymatic study of newly synthesized analogs of trypsin inhibitors SFTI-1 that were fluorescent labeled on their N-terminal amino groups. Two fluorescent derivatives of benzoxazole (3-[2-(4-diphenylaminophenyl) benzoxazol-5-yl]-L-alanine[(4NPh(2)) Ph] Box-Ala and 3-[2-(2', 4', 5'-trimethoxyphenyl) benzoxazol-5-yl]-l-alanine-[2,4,5-(OMe)3Ph] Box-Ala) were used as efficient fluorescent labels....
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Glucosamine-6-phosphate synthase with an oligoHis insert
PublicationGlucosamine-6-phosphate (GlcN-6-P) synthase known also as L-Glutamine: D-fructose-6-phosphate aminotransferase (EC 2.6.1.16), catalyzes the first committed step in the amino sugar biosynthetic pathway in prokaryotic and eukaryotic organisms. The final product of this pathway is an activated precursor of numerous macromolecules containing amino sugars, including chitin and mannproteins in fungi, peptydoglican and lipopolysaccharides...
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Synergic Effect of Naphthylated Carbon Nanotubes and Gold Nanoparticles on Catalytic Performance of Hybrid Films Containing Bilirubin Oxidase for the Dioxygen Reduction
PublicationTo improve the direct electron transfer from a multicopper oxidase MvBOd to the glassy carbon electrode surface the naphthylated multiwall carbon nanotubes (Napht-MWCNTs) are ombined with gold nanoparticles (AuNPs) to form a conductive network at the electrode and to increase its working surface. Such approach allowed to eliminate the necessity of employing mediators to reach elevated current values. The enzyme stability was improved...
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Characterization of immobilized Escherichia coli cells transformed by β-galactosidase gene from Pyrococcus woesei
PublicationThermostable β-galactosidase from Escherichia coli transformant containing theenzyme gene from Pyrococcus woesei was immobilized on alginate gel.The benefits of using whole bacterial cells not only exclude expensive, laborious proteinisolation and purification but also stabilize enzymes by cytosol components. Increase inproductivity of enzyme can be achieved by cells permeabilization. To increase permeability ofcytoplasmic membrane...
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Catalytic topoisomerase II inhibitors in cancer therapy.
PublicationDNA topoizomeraza II jest jednym z głównych celów molekularnych dla leków stosowanych w terapii przeciwnowotworowej. Wszystkie inhibitory tego enzymu są zdolne do zakłócania w jeden z etapów cyklu katalitycznego topoizomerazy II. Związki, które stabilizują kowalencyjny kompleks DNa i topoizomerazy II (znany jako kompleks rozszczepialny) są nazywane truciznami topoizomerazy, inne związki, które hamują aktywność enzymu na innych...
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Comparison of oil yield and quality obtained by different extraction procedures from salmon (Salmo salar) processing byproducts
PublicationThe content and composition of lipids in different byproducts (skins, heads, and backbones) from mechanically processed farmed Atlantic salmon were determined and compared with that obtained from wild salmon. Three different procedures were used to establish the optimal conditions of oil extraction (at high temperature −95°C, “cold” extraction at temperature not exceeding 15°C and enzyme assisted with Alcalase). “Cold” extraction...
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Role of cholesterol in substrate recognition by -secretase
Publication-Secretase is an enzyme known to cleave multiple substrates within their transmembrane domains, with the amyloid precursor protein of Alzheimer’s Disease among the most prominent examples. The activity of -secretase strictly depends on the membrane cholesterol content, yet the mechanistic role of cholesterol in the substrate binding and cleavage remains unclear. In this work, we used all-atom molecular dynamics simulations to examine...
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Molecular characterization of a cryptic plasmid from the psychrotrophic antarctic bacterium Pseudoalteromonas sp. 643A
PublicationW ostatnich latach znacznie wzrosło zainteresowanie organizmami zimnolubnymi, głównie z powodu ich enzymów o biotechnologicznym znaczeniu. Jednak, aby wydajnie produkować takie enzymy w skali przemysłowej użyteczne jest stworzenie rekombinantowych systemów ekspresyjnych, które pozwolą na ich opłacalną produkcję. W osiągnięciu tego celu mogą pomóc odpowiednie wektory, konstruowane na bazie plazmidów pozyskiwanych z bakterii psychrofilnych/psychrotrofowych....
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Sławomir Milewski prof. dr hab. inż.
PeopleSławomir Milewski, born in 1955 in Puck, Poland, graduated in 1979 in Chemistry from the Faculty of Chemistry, Gdańsk University of Technology (GUT). In 1984 was employed at the Department of Pharmaceutical Technology and Biochemistry. In 1985 he got his PhD, in 1994 became a DSc (habilitation) and in 2002 got the professorship in chemical sciences. Currently he is a full professor and Head of the Department of Pharmaceutical Technology...
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Isolation and some properties of the thermostable beta-galactosidase of Pyrococcus woesei expressed in Escherichia coli
PublicationOpracowano metodę izolowania oraz zbadano właściwości enzymu otrzymanego ze szczepu Escherichia coli zawierającego gen kodujący termostabilną beta-ga-laktozydazę z Pyrococcus woesei.
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Marta Wanarska dr hab. inż.
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Oxygen biosensor based on carbon nanotubes directly grown on graphitic substrate
PublicationFilms of carbon nanotubes were synthesized on the graphitic substrates by CVD method in the presence of catalyst. Two different catalysts were used, that is nickel and iron, supported on graphitic substrates. The as-obtained nanotubes were analyzed by SEM imaging with EDX and Raman spectroscopy. The results revealed their multi-walled morphology. Free radical reaction was carried out in order to chemically functionalize synthesized...
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In silico design of telomerase inhibitors.
PublicationTelomerase is a reverse transcriptase enzyme involved in DNA synthesis at the end of linear chromosomes. Unlike in most other cells, telomerase is reactivated most cancerous cells and, therefore, has become a promising new anticancer target. Despite extensive research, direct telomerase inhibitors have yet not been introduced to the clinics because of the complexity of this enzyme. Structures of this protein from simple organisms...
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Regulacja aktywności syntazy glukozamino-6-fosforanu z Candida albicans – badania metodami obliczeniowymi
PublicationNiniejsza praca przedstawia wyniki badań, prowadzonych metodami obliczeniowymi, poświęconych enzymowi syntazie GlcN-6-P pochodzącemu z drożdży C. albicans. Enzym ten odpowiedzialny jest między innymi za biosyntezę kluczowych składników ściany komórkowej grzybów i bakterii, ma również wpływ na regulację metabolizmu glukozy. Przedstawione badania stanowią jeden z etapów poszukiwania molekularnego mechanizmu inhibicji, której eukariotyczna...
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High level expression, secretion and purification of the termostable aqualysin I from Thermus aquaticus YT-1 in Pichia pastoris.
PublicationOtrzymano aktywną proteazę serynową Thermus aquaticus z systemu nadprodukcji Escherichia coli. Oczyszczono enzym przy zastosowaniu chromatografii metalo-powinowactwa.
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Comparative molecular dynamics study of dimeric and monomeric forms of HIV-1 protease in ligand bound and unbound state
PublicationHuman immunodeficiency virus type 1 protease (HIV-1 PR) is a viral-encoded enzyme that forms a homodimer. HIV-1 PR is essential for replication and assembly of the virus and inactivation of HIV-1 PR enzyme causes production of immature, noninfectious viral particles and thus HIV-1 PR is an attractive target in anti-AIDS drug design. In our current work, we performed molecular dynamics (MD) calculations (500 ns) for two different...
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Modulation of UDP-glucuronidation by acridinone antitumor agents C-1305 and C-1311 in HepG2 and HT29 cell lines, despite slight impact in noncellular systems.
PublicationBackground Among the studied antitumor acridinone derivatives developed in our laboratory, 5-dimethylaminopropylamino-8-hydroxytriazoloacridinone (C-1305) and 5-diethylaminoethylamino-8-hydroxyimidazoacridinone (C-1311) exhibited cytotoxic and antitumor properties against several cancer types and were selected to be evaluated in preclinical and early-phase clinical trials. In the present work, we investigated the impact of C-1305...
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Active Site Architecture and Reaction Mechanism Determination of Cold Adapted beta-D-galactosidase from Arthrobacter sp. 32cB
PublicationArthbetaDG is a dimeric, cold-adapted beta-D-galactosidase that exhibits high hydrolytic and transglycosylation activity. A series of crystal structures of its wild form, as well as its ArthbetaDG_E441Q mutein complexes with ligands were obtained in order to describe the mode of its action. The ArthbetaDG_E441Q mutein is an inactive form of the enzyme designed to enable observation of enzyme interaction with its substrate. The...
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Plasmids pMET2M1 and pMET15M1 with the SAT1 Flipper knockout cassette construction
Open Research DataThe goal of the study was to obtain plasmids containing the SAT1 Flipper knockout cassette along with upstream and downstream gene fragments (MET2, MET15) and to introduce a cleavage site for restriction enzymes into them.
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Structure of EstA esterase from psychrotrophic Pseudoalteromonas sp. 643A covalently inhibited by monoethylphosphonate.
PublicationThe crystal structure of the esterase EstA from the cold-adapted bacteriumPseudoalteromonas sp. 643A was determined in a covalently inhibited form at aresolution of 1.35 A˚. The enzyme has a typical SGNH hydrolase structureconsisting of a single domain containing a five-stranded beta-sheet, with threehelices at the convex side and two helices at the concave side of the sheet, and isornamented with a couple of very short helices...
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Antifungal Activity of Homoaconitate and Homoisocitrate Analogs
PublicationThirteen structural analogs of two initial intermediates of the L-alpha-aminoadipate pathway of L-lysine biosynthesis in fungi have been designed and synthesized, including fluoro- and epoxy-derivatives of homoaconitate and homoisocitrate. Some of the obtainedcompounds exhibited at milimolar range moderate enzyme inhibitory properties against homoaconitase and/or homoisocitrate dehydrogenase of Candida albicans. The structural...
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Imidazoacridinone antitumor agent, c-1311, as a selective mechanism-based inactivator of human recombinant cytochrome P450 1A2
PublicationPrzedmiotem badań była przeciwnowotworowa pochodna imidazoakrydonu, C-1311, która pod nazwą Symadex R, znajduje się aktualnie w II fazie badań klinicznych. Wcześniejsze wyniki pozwoliły stwierdzić, że związek ten jest selektywnym inhibitorem niektórych enzymów cytochromowych. Podjęto zatem próbę wyjaśnienia mechanizmu inhibicji izoenzymów cytochromu P450 przez C-1311. Wykazano, że utrata aktywności katalitycznej obu izoenzymów...
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Beta-galactosidases from a psychrotolerant Arthrobacter isolates and their potential use
PublicationEnzymes from extremophiles, especially psychrophiles and psychrotolerant microorganisms are currently intensively studied because of their unusual structural and functional properties. One of these enzymes is a cold-adapted - galactosidase (EC 3.2.1.23) which catalyzes the hydrolysis of - 1,4-glycosidic bonds in -galactosides. The most common - galactoside is lactose, a disaccharide which makes up 2-7% of milk. The majority of...
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Desensitization of glucosamine-6-phosphate synthase to inhibition by UDP-GlcNAc
PublicationGlucosamine-6-phosphate (GlcN-6-P) synthase, known also as L-Glutamine: D-fructose-6-phosphate amidotransferase, catalyzes the first committed step in the pathway leading to the ultimate formation of UDP-GlcNAc. The final product of this pathway is an activated precursor of numerous macromolecules containing amino sugars, including chitin and mannproteins in fungi, peptidoglycan and lipopolysaccharides in bacteria, and glycoproteins...
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Structural studies of a cold-adapted dimeric Beta-D-galactosidase from Paracoccus sp. 32d
PublicationThe crystal structure of a novel dimeric [beta]-D-galactosidase from Paracoccus sp. 32d (Par[beta]DG) was solved in space group P212121 at a resolution of 2.4 Å by molecular replacement with multiple models using the BALBES software. This enzyme belongs to glycoside hydrolase family 2 (GH2), similar to the tetrameric and hexameric [beta]-D-galactosidases from Escherichia coli and Arthrobacter sp. C2-2, respectively. It is the second...
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Inserts amplification for knockout cassette construction
Open Research DataThe goal of the study was to obtain optimal conditions for amplification of upstream and downstream genes fragments and introduce a cleavage site for restriction enzymes into them thanks to which it will be possible to clone fragments into a plasmid containing elements of the knockout cassette. The constructed cassettes will enable the removal of the...
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Increased concentration of Taq DNA polymerase as a solution for GC-rich templates from clinical and environmental samples
PublicationDNA polymerase is an enzyme which plays crucial role in replication and DNA repair. It found application in PCR (polymerase chain reaction) where catalyses process of in vitro DNA synthesis. To meet the demands posed by mod- ern diagnostic, molecular biology or genetic engineering it is necessary to improve DNA polymerases to obtain new or better features useful in these fields. So far implemented modifications in majority are...
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Catalytic Mechanism of Non-Target DNA Cleavage in CRISPR-Cas9 Revealed by Ab Initio Molecular Dynamics
PublicationCRISPR-Cas9 is a cutting-edge genome editing technology, which uses the endonuclease Cas9 to introduce mutations at desired sites of the genome. This revolutionary tool is promising to treat a myriad of human genetic diseases. Nevertheless, the molecular basis of DNA cleavage, which is a fundamental step for genome editing, has not been established. Here, quantum–classical molecular dynamics (MD) and free energy methods are used...