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Wyniki wyszukiwania dla: MILK PROTEIN ALLERGENICITY
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Pilicides effectively hinder chaperone-usher interactions in all Gram-negative bacteria
PublikacjaThe chaperone-usher pathway of adhesive structures biogenesis is highly conserved in all Gram-negative bacteria. Pili are polymers of thousands protein subunits that possess conserved immunoglobuline-like structure denoted by lack of the seventh G strand. The effect of this structural defect is a hydrophobic acceptor cleft. The folding of protein subunits is strictly dependent on the action of specific periplasmic chaperone protein...
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Fusion of DNA-binding domain of Pyrococcus furiosus ligase with TaqStoffel DNA polymerase as a useful tool in PCR with difficult targets
PublikacjaThe DNA coding sequence of TaqStoffel polymer- ase was fused with the DNA-binding domain of Pyrococcus furiosus ligase. The resulting novel recombinant gene was cloned and expressed in E. coli. The recombinant enzyme was purified and its enzymatic features were studied. The fusion protein (PfuDBDlig-TaqS) was found to have enhanced processivity as a result of the conversion of the Taq DNA polymerase from a relatively low processive...
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A novel cold-active β-D-galactosidase with transglycosylation activity from the Antarctic Arthrobacter sp. 32cB - gene cloning, purification and characterization
PublikacjaA gene encoding a novel β-D-galactosidase from the psychrotolerant Antarctic bacterium Arthrobacter sp. 32cB was isolated, cloned and expressed in Escherichia coli. The active form of recombinant β-D-galactosidase consists of two subunits with a combined molecular weight of approximately 257 kDa. The enzyme's maximum activity towards o-nitrophenyl-β-D-galactopyranoside was determined as occurring at 28 °C and pH 8.0. However, it...
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Extracting functional groups of ALLINI to design derivatives of FDA‐approved drugs: Inhibition of HIV‐1 integrase
PublikacjaHIV‐1 integrase (IN) is crucial for integration of viral DNA into the host genome and a promising target in development of antiretroviral inhibitors. In this work, six new compounds were designed by linking the structures of two different class of HIV‐1 IN inhibitors (active site binders and allosteric IN inhibitors (ALLINIs)). Among newly designed compounds, INRAT10b was found most potent HIV‐1 IN inhibitor considering different...
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Synthesis and SAR Studies of Antibacterial Peptidyl Derivatives Based upon the Binding Site of Human Cystatin C
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Construction, purification, and functional characterization of His-tagged Candida albicans glucosamine-6-phosphate synthase expressed in Escherichia coli
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Purification to Homogeneity of Candida albicans Glucosamine-6-phosphate Synthase Overexpressed in Escherichia coli
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Analogues of Trypsin Inhibitor SFTI-1 with Disulfide Bridge Substituted by Various Length of Carbonyl Bridges
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A two-plasmid Escherichia coli system for expression of Dr adhesins
PublikacjaPraca opisuje konstrukcję wydajnego systemu produkcji fimbrii Dr. System składa się z dwóch plazmidów. Plazmid pACYCpBAD-DraC-C-His zawiera pod kontrolą promotora arabinozowego gen draC kodujący białko kanałotwórcze DraC. Plazmid pET-30b-sygDraBE zawiera pod kontrolą promotora T7lac geny draB i draE kodujące odpowiednio białko opiekuńcze DraB oraz adhezynę DraE. Oba plzamidy posiadają różne ori replikacji co pozwala na ich jednoczesne...
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Molecular Simulations of Rhodopsin Tetrameter
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Therapy-Related Changes in the Serum Proteome Patterns of Early Stage Breast Cancer Patients with Different Outcomes
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Molecular Modeling of the Neurohypophyseal Receptor/Atosiban Complexes
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Molecular Dynamics Study of the Internal Water Molecules in Vasopressin and Oxytocin Receptors
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Low-Molecular-Weight Aldehyde Inhibitors of Cathepsin G
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Pegylated Fluorescent Peptides as Substrates of Proteolytic Enzymes
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Designing of Substrates and Inhibitors of Bovine α-Chymotrypsin with Synthetic Phenylalanine Analogues in Position P1
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Novel method of expression and purification of hirudin based on pBAD TOPO, pTYB12 vectors and gene synthesis
PublikacjaW pracy przedstawiono nową, szybką i tanią metodę otrzymywania, ekspresji i oczyszczania białka hirudyny naturalnie izolowanego z ślinianek pijawki lekarskiej. Hirudyna pełni funkcje bezpośredniego inhibitora trombiny, głównego białka biorącego udział w tworzeniu skrzepów krwi. Właściwości te wykorzystywane są od wielu lat w klinice. Opracowana metoda pozwoliła uzyskać 17 mg/l hodowli E. coli białka o wysokiej czystości i aktywności...
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Extracellular secretion of Pseudoalteromonas sp. cold-adapted esterase in Escherichia coli in the presence of Pseudoalteromonas sp. components of ABC transport system
PublikacjaEnzymy lipolityczne, głównie lipazy i esterazy, stanowią cenną grupę biokatalizatorów wykorzystywanych w przemyśle. Coraz większe zainteresowanie tą klasą enzymów sprawiło, że poszukuje się alternatywnych źródeł ich pozyskiwania. W celu zbadania wpływu białek transportu ABC Pseudoalteromonas sp. 643A na ekspresję genu, sekrecję i aktywność esterazy EstA z tego mikroorganizmu w systemie ekspresyjnym E. coli skonstruowano układ do...
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Novel thermostable ssDNA-binding proteins from Thermus thermophilus and T. aquaticus-expression and purification
PublikacjaSklonowano w system ekspresyjny pET-30LIC Escherichia coli gen kodujący białka SSB-podobne Thermus thermophilus i T. aquaticus. Oczyszczono do homogenności białka SSB-podobne, a uzyskane preparaty umożliwiają wykonanie badań strukturalnych w/w białek.
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High yield expression and single step purification of Toxoplasma gondii SAG1, GRA1, GRA7 antigens in Escherichia coli.
PublikacjaPraca przedstawia metodę uzyskiwania antygenów rekombinowanych Toxoplasmagondii o wysokiej czystości, które można zastosować w immunodiagnostyce. Antygeny T. gondii eksprymowane w komórkach E. coli zawierały polihistydynowedomeny fuzyjne na końcu N i C białka. Pozwalało to na jednoetapowe oczyszczanie białek metodą chromatografii metalopowinowactwa na złożu Ni2+ -IDA-Sepharose. Immunoreaktywność antygenów rekombinowanych...
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High level expression, secretion and purification of the termostable aqualysin I from Thermus aquaticus YT-1 in Pichia pastoris.
PublikacjaOtrzymano aktywną proteazę serynową Thermus aquaticus z systemu nadprodukcji Escherichia coli. Oczyszczono enzym przy zastosowaniu chromatografii metalo-powinowactwa.
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Characterization of recombinant homocitrate synthase from Candida albicans
PublikacjaLYS21 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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Introduction of Pro and Its Analogues in the Conserved P1 Position of Trypsin Inhibitor SFTI-1 Retains Its Inhibitory Activity
PublikacjaA number of monocyclic SFTI-1 analogues modified in the conserved inhibitor P1 position by Pro, its L-hydroxyproline (Hyp) derivative as well as mimetics with different ring size were synthesized by the solid-phase method. Replacement of Ser6 by Pro, Hyp, and a four-member ring, L-azetidine-2-carboxylic acid (Aze), retained trypsin or chymotrypsin inhibitory activity. The determined association equilibrium constants of these analogues...
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New conjugates of tuftsin and muramyl dipeptide as stimulators of human monocytes-derived dendritic cells
PublikacjaMuramyl dipeptide (MDP) and tuftsin are known biologically active compound displaying a significant influence on various cell populations of innate immune response. MDP, as a fragment of bacterial cell wall, stimulates not only macrophages and monocytes, but also dendritic cells. In contrast, little is known about tuftsin influence on these cells. Therefore it seemed vital to access whether tuftsin or its derivatives conjugated...
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DNA-reactive anticancer imidazoacridinone C-1311 is a new inhibitor of hypoxia-inducible factor 1 alpha, vascular endothelial growth factor and tumor angiogenesis
PublikacjaHypoxia-inducible factor 1 (HIF-1) plays a critical role for tumor adaptation to hypoxia and promotes angiogenesis. Antitumor imidazoacridinone C-1311 is a DNA reactive topoisomerase II and FLT3 receptor tyrosine kinase inhibitor selected for phase II clinical trials for breast cancer. Here, we demonstrate the new mechanism of C-1311 action involving HIF-1a, vascular endothelial growth factor (VEGF) and angiogenesis as additional...
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Mechanism of Osmolyte Stabilization–Destabilization of Proteins: Experimental Evidence
PublikacjaIn this work, we investigated the influence of stabilizing (N,N,N-trimethylglycine) and destabilizing (urea) osmolytes on the hydration spheres of biomacromolecules in folded forms (trpzip-1 peptide and hen egg white lysozyme─hewl) and unfolded protein models (glycine─GLY and N-methylglycine─NMG) by means of infrared spectroscopy. GLY and NMG were clearly limited as minimal models for unfolded proteins and should be treated with...
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Triazoloacridone C-1305 impairs XBP1 splicing by acting as a potential IRE1α endoribonuclease inhibitor
PublikacjaInositol requiring enzyme 1 alpha (IRE1α) is one of three signaling sensors in the unfolding protein response (UPR) that alleviates endoplasmic reticulum (ER) stress in cells and functions to promote cell survival. During conditions of irrevocable stress, proapoptotic gene expression is induced to promote cell death. One of the three signaling stressors, IRE1α is an serine/threonine-protein kinase/endoribonuclease (RNase) that...
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Scarabaeidae as human food – A comprehensive review
PublikacjaRising global population and sustainable protein demand have sparked interest in unique food sources. Entomophagy, or insect consumption, presents a solution and Scarab beetles, part of the Scarabaeidae family, offer a novel food option. The comprehensive review underscores their potential as human food, with strong nutrition, low environmental impact, and the ability to ease strain on conventional agriculture. Nutritional analysis...
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Preparation and characterization of porous scaffolds from chitosan-collagen-gelatin composite
PublikacjaNovel porous scaffolds composed of chitosan, collagen and gelatin were prepared and characterized. For preparing scaffolds gelatin and collagen isolated from fish skins with various physicochemical properties were used. In order to reduce preparation solubility glutaraldehyde in the amount of 1%, w/w relative to the total biopolymers weight in solution was used. All obtained biomaterials showed a homogeneous porosity. Protein polymer...
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Hydration of Simple Model Peptides in Aqueous Osmolyte Solutions
PublikacjaThe biology and chemistry of proteins and peptides are inextricably linked with water as the solvent. The reason for the high stability of some proteins or uncontrolled aggregation of others may be hidden in the properties of their hydration water. In this study, we investigated the effect of stabilizing osmolyte–TMAO (trimethylamine N-oxide) and destabilizing osmolyte–urea on hydration shells of two short peptides, NAGMA (N-acetyl-glycine-methylamide)...
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Early Stages of RNA-Mediated Conversion of Human Prions
PublikacjaPrion diseases are characterized by the conversion of prion proteins from a PrPC fold into a disease-causing PrPSC form that is self-replicating. A possible agent to trigger this conversion is polyadenosine RNA, but both mechanism and pathways of the conversion are poorly understood. Using coarse-grained molecular dynamic simulations we study the time evolution of PrPC over 600 μs. We find that both the D178N mutation and interacting...
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Hopf bifurcation in time‐delayed gene expression model with dimers
PublikacjaWe study a mathematical model of gene transcription and protein synthesis with negative feedback. We consider a system of equations taking into account the formation of dimers (i.e., complex formed by two protein monomers), the way in which dimers bind to DNA and time delay in translation process. For the model consisting of three ordinary differential equations with time delay, we derive conditions for stability of the positive...
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Correlation between the number of Pro-Ala repeats in the EmrA homologue of Acinetobacter baumannii and resistance to netilmicin, tobramycin, imipenem and ceftazidime
PublikacjaAcinetobacter baumannii coccobacilli are dangerous to patients in intensive care units because of their multidrug resistance to antibiotics, developed mainly in the past decade. This study aimed to examine whether there is a significant correlation between the number of Pro-Ala repeats in the CAP01997 protein, the EmrA homologue of A. baumannii, and resistance to antibiotics. A total of 79 multidrug-resistant A. baumannii strains...
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Cloning, expression, and biochemical characterization of a coldactive GDSL-esterase of a Pseudomonas sp. S9 isolated from Spitsbergen island soil
PublikacjaAn estS9 gene, encoding an esterase of the psychrotolerant bacterium Pseudomonas sp. S9 was cloned and sequenced. The deduced sequence revealed a protein of 636 amino acid residues with a molecular mass of 69 kDa.Further amino acid sequence analysis revealed that the EstS9 enzyme contained a G-D-S-L motif centered at a catalytic serine, an N-terminal catalytic domain and a C-terminal autotransporter domain. Two recombinant E. coli...
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Microencapsulation of fish oil – determination of optimal wall material and encapsulation methodology
PublikacjaFor the first time, we present a meta-analysis of experimental and literature data to determine which microencapsulation methodology, and which wall material are best suited to protect fish oil. Our analysis covered a period of several decades of research (1984–2018). The analysis was conducted on 196 literature data-points, and 16 data-points determined experimentally for this publication. PLS regression was used to determine...
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Interfacial water controls the process of adsorption of hyperactive antifreeze proteins onto the ice surface
PublikacjaA mechanism of interactions between the ice-binding surface of a hyperactive antifreeze protein molecule and the ice surface is proposed, involving the influence of water present between the two surfaces on the behavior of the approaching molecule. It is demonstrated that the interfacial water, even before its full solidification, can act as a factor that pushes away or pulls nearer the protein molecule to ensure its proper positioning....
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Effect of conventional cooking on changes in the contents of basic composition and glucosinolates in kale
PublikacjaBrassica vegetables have been strongly recommended as part of human diet because of its high content of bioactive sulphur compounds, eg glucosinolates. The nutrient and health promoting compounds in kale are significantly affected by traditional cooking. The study investigated changes in the levels of dry mass, ash, fat, total protein, dietary fibre as well as total and individual glucosinolates in the kale due to the traditional...
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A novel cold-active beta-D-galactosidase from the Paracoccus sp. 32d - gene cloning, purification and characterization
PublikacjaBeta-D-galactosidase (EC 3.2.1.23) catalyze the hydrolysis of terminal non-reducing beta-D-galactose residues in beta-D-galactosides. Cold-active beta-D-galactosidases have recently become a focus of attention of researchers and dairy product manufactures owing to theirs ability to: (I) eliminate of lactose from refrigerated milk for people afflicted with lactose intolerance, (II) convert lactose to glucose and galactose which...
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3-MCPD: A worldwide problem of food chemistry
Publikacja3-MCPD is a heat-induced food contaminant which has been widely investigated for decades. This paper presents an overview of current knowledge about 3-MCPD including its formation routes, occurrence in various foodstuffs, analytical approach, toxicological aspects and future research perspectives. So far 3-MCPD was determined in its free and bound form in thermally-treated foods, edible oils and fats, and infant foods including...
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Data regarding a new, vector-enzymatic DNA fragment amplification-expression technology for the construction of artificial, concatemeric DNA, RNA and proteins, as well as biological effects of selected polypeptides obtained using this method
PublikacjaApplications of bioactive peptides and polypeptides are emerging in areas such as drug development and drug delivery systems. These compounds are bioactive, biocompatible and represent a wide range of chemical properties, enabling further adjustments of obtained biomaterials. However, delivering large quantities of peptide derivatives is still challenging. Several methods have been developed for the production of concatemers –...
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Which casein in sodium caseinate is most resistant to in vitro digestion? Effect of emulsification and enzymatic structuring
PublikacjaWe investigated the resistance of individual constituent casein epitopes (αS1-, αS2-, β- and κ-CN) in food-grademilk protein sodium caseinate (NaCN) to simulated human gastro-duodenal digestion. The influence of NaCNadsorption to the surface of oil-in-water emulsion droplets and the effect of crosslinking of the protein withenzyme transglutaminase (TG) on the proteolysis were studied by indirect ELISA. TG crosslinking renderedfragments...
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Example of utilization of permeabilized microbial cells
PublikacjaThe benefits of using whole bacterial cells not only exclude expensive, laborious protein isolation and purification but also stabilize enzymes by cytosol components. Increase in activity of the cells can be achieved by cells permeabilization.
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Investigation of interaction between xanthene dye rhodamine B and metagenomic-derivated methylthioadenosine phosphorylase RSFP
PublikacjaIn the previous study we found that the expression of the metagenomic - derived methylthioadenosine phosphorylase gene (rsfp) in E. coli cells and the presence of rhodamine B in the growth medium were crucial for the pink fluorescence of one E. coli colony from Antarctic soil metagenomic library (Cieśliński et al., 2009). In this study, rsfp gene expression was induced by an arabinose promoter in an LMG194 E. coli strain and the...
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Properties of water in the region between a tubulin dimer and a single motor head of kinesin
PublikacjaA kinesin is a molecular motor that can perform movement on a microtubule track in a stepping-like manner. This motion is connected with processes of association and dissociation of kinesin and tubulin. Water is an important participant in these kinds of molecular interactions. This is why we have decided to investigate the dynamical and structural properties of water in the region between the kinesin catalytic domain and the tubulin...
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Biochemical characteristic of biofilm of uropathogenic Escherichia coli Dr+ strains.
PublikacjaUrinary tract infections caused by Escherichia coli are very common health problem in the developed countries. The virulence of the uropathogenic E. coli Dr+ IH11128 is determined by Dr fimbriae, which are homopolymeric structures composed of DraE subunits with the DraD protein capping the fiber. In this study, we have analyzed the structural and biochemical properties of biofilms developed by E. coli strains expressing Dr fimbriae...
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Why the Solvation Water around Proteins Is More Dense than Bulk Water
PublikacjaThe main aim of this work is to propose a rational explanation of the commonly observed phenomenon of increasing water density within solvation shell of proteins. We have observed that the geometry of the water–water hydrogen bond network within solvation layer differs from the one in bulk water, and it is the result of interactions of water molecules with protein surface. Altered geometry of the network reflects changes in the...
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Why the solvation water around proteins is more dense than bulk water
PublikacjaThe main aim of this work is to propose a rational explanation of commonly observed phenomenon of increasing water density within solvation shell of proteins. We have observed that geometry of the water-water hydrogen bond network within solvation layer differs from the one in bulk water and it is the effect of interactions of water molecules with protein surface. Altered geometry of the network reflects changes in the structure...
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Identification and properties of the psychrophilic bacterium Desulfolatea psychrophila single-stranded DNA binding proteins
PublikacjaTo study the biochemical properties of SSB from Desulfolatea psychrophila (DpsSSB), we have cloned the ssb genes obtained by PCR and have developed Escherichia coli overexpression systems. The gene consists of an open reading frame of 426 nucleotides encoding SSB protein of 142 amino acids with a calculated molecular mass of 15.6 kDa and it is the smallest known bacterial SSB protein indentified to date. The amino-acid sequence...
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Interactions of telomeric proteins with nucleic acids: sequence recognition on intact and oxidatively damaged telomeres
PublikacjaTelomeres are complex nucleoprotein assemblies that play a vital role in the maintenance of functional ends of linear chromosomes. Telomeric DNA, composed of tandem repeats of the 5'-TTAGGG-3' motif, solves the so-called end replication problem: as chromosomes shorten with each cell division, no information is lost, and the telomere can be re-extended. In the cell, many protein factors regulate telomere length, nuclear positioning...
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Effects of the polyhistidine tag on kinetics and other properties of trehalose synthase from Deionococcus geothermalis
PublikacjaTwo recombinant trehalose synthases from Deinococcus geothermalis (DSMZ 11300) were compared. A significant influence of the artificial polyhistidine tag was observed in protein constitution. The recombinant trehalose synthase from D. geothermalis with His6 -tag has a higher K m value of 254 mM, in comparison with the wild-type trehalose synthase (K m 170 mM), and displayed a lower activity of maltose conversion when compared...