scholarly journals Investigating the Product Profiles and Structural Relationships of New Levansucrases with Conventional and Non-Conventional Substrates

2020 ◽  
Vol 21 (15) ◽  
pp. 5402
Author(s):  
Andrea Hill ◽  
Salwa Karboune ◽  
Tarun J. Narwani ◽  
Alexandre G. de Brevern
Keyword(s):  
Food Industry ◽  
Genome Mining ◽  
Gluconobacter Oxydans ◽  
Amino Acid Sequences ◽  
Structural Elements ◽  
Initial Screening ◽  
Donor Molecule ◽  
Structural Relationships ◽  
High Degree ◽  
Homology Models

The synthesis of complex oligosaccharides is desired for their potential as prebiotics, and their role in the pharmaceutical and food industry. Levansucrase (LS, EC 2.4.1.10), a fructosyl-transferase, can catalyze the synthesis of these compounds. LS acquires a fructosyl residue from a donor molecule and performs a non-Lenoir transfer to an acceptor molecule, via β-(2→6)-glycosidic linkages. Genome mining was used to uncover new LS enzymes with increased transfructosylating activity and wider acceptor promiscuity, with an initial screening revealing five LS enzymes. The product profiles and activities of these enzymes were examined after their incubation with sucrose. Alternate acceptor molecules were also incubated with the enzymes to study their consumption. LSs from Gluconobacter oxydans and Novosphingobium aromaticivorans synthesized fructooligosaccharides (FOSs) with up to 13 units in length. Alignment of their amino acid sequences and substrate docking with homology models identified structural elements causing differences in their product spectra. Raffinose, over sucrose, was the preferred donor molecule for the LS from Vibrio natriegens, N. aromaticivorans, and Paraburkolderia graminis. The LSs examined were found to have wide acceptor promiscuity, utilizing monosaccharides, disaccharides, and two alcohols to a high degree.

Cloning of the Rat Tissue Factor cDNA and Promoter: Identification of a Serum-response Region

1996 ◽  
Vol 76 (05) ◽  
pp. 697-702 ◽  
Author(s):  
Olivier Taby ◽  
Claire-Lise Rosenfield ◽  
Vladimir Bogdanov ◽  
Yale Nemerson ◽  
Mark B Taubman
Keyword(s):  
Tissue Factor ◽  
General Structure ◽  
Amino Acid Sequences ◽  
Proximal Promoter ◽  
Striking Similarity ◽  
Upstream Sequence ◽  
Tf Gene ◽  
Rat Tissue ◽  
High Degree ◽  
Transcriptional Elements

SummaryTissue factor (TF) initiates coagulation and its expression in vascular smooth muscle cells (VSMC) likely plays a role in the propagation of arterial thrombosis. We report cloning the cDNA and proximal promoter region of the rat TF gene. While maintaining the general structure and organization of the TF molecule, there is a surprising divergence (≈ 18%) between the derived amino acid sequences of the rat and mouse TF. In contrast, there is striking similarity (90%) in the 5’ untranslated regions. High levels of basal promoter activity were seen in rat VSMC with constructs containing 106 bp of sequence downstream from the putative transcription start site and 426 to 103 bp of upstream sequence. Deletion of the sequence from −103 to −79, containing a single SP1 site, removed virtually all of the basal and serum-induced activity. Removal of the NFkB site or two additional upstream SP1 sites had little effect on serum responsiveness. Removal of the 5’ untranslated region abolished most of the basal activity of the TF promoter, suggesting that its high degree of conservation may be due to the presence of transcriptional elements critical for TF expression in rodent VSMC.

scholarly journals Structure and expression of human IgG FcRII(CD32). Functional heterogeneity is encoded by the alternatively spliced products of multiple genes.

1989 ◽  
Vol 170 (4) ◽  
pp. 1369-1385 ◽  
Author(s):  
D G Brooks ◽  
W Q Qiu ◽  
A D Luster ◽  
J V Ravetch
Keyword(s):  
Structural Heterogeneity ◽  
Amino Acid Sequences ◽  
Cdna Clones ◽  
Membrane Glycoproteins ◽  
Igg Binding ◽  
Genes Expression ◽  
Alternatively Spliced ◽  
Membrane Spanning ◽  
High Degree ◽  
Membrane Spanning Domains

The structural heterogeneity of the human low affinity receptor for IgG, FcRII(CD32), has been elucidated through the isolation, characterization, and expression of cDNA clones derived from myeloid and lymphoid RNA. These clones predict amino acid sequences consistent with integral membrane glycoproteins with single membrane spanning domains. The extracellular domains display sequence homology to other Fc gamma Rs and members of the Ig supergene family. A minimum of three genes (Fc gamma RIIa, IIa', and Fc gamma RIIb) encode these transcripts, which demonstrate highly related extracellular and membrane spanning domains. IIa/IIa' differ substantially in the intracytoplasmic domain from IIb. Alternative splicing of the IIb gene generates further heterogeneity in both NH2- and COOH-terminal domains of the predicted proteins. Comparison to the murine homologues of these molecules reveals a high degree of conservation between the products of one of these genes, Fc gamma RIIb, and the murine beta gene in primary sequence, splicing pattern, and tissue distribution. In contrast, the sequence of IIa' indicates its relationship to the beta-like genes, with mutation giving rise to a novel cytoplasmic domain, while IIa is a chimera of both alpha- and beta-like genes. Expression of these cDNA molecules by transfection results in the appearance of IgG binding molecules that bear the epitopes defined by the FcRII(CD32) mAbs previously described.

scholarly journals The TFIID Components Human TAFII140 andDrosophila BIP2 (TAFII155) Are Novel Metazoan Homologues of Yeast TAFII47 Containing a Histone Fold and a PHD Finger

2001 ◽  
Vol 21 (15) ◽  
pp. 5109-5121 ◽  
Author(s):  
Yann-Gaël Gangloff ◽  
Jean-Christophe Pointud ◽  
Sylvie Thuault ◽  
Lucie Carré ◽  
Christophe Romier ◽  
...  
Keyword(s):  
Rna Polymerase Ii ◽  
Sequence Similarity ◽  
Protein A ◽  
Amino Acid Sequences ◽  
Molecular Organization ◽  
Phd Finger ◽  
Histone Fold ◽  
Histone Fold Domain ◽  
High Degree

ABSTRACT The RNA polymerase II transcription factor TFIID comprises the TATA binding protein (TBP) and a set of TBP-associated factors (TAFIIs). TFIID has been extensively characterized for yeast, Drosophila, and humans, demonstrating a high degree of conservation of both the amino acid sequences of the constituent TAFIIs and overall molecular organization. In recent years, it has been assumed that all the metazoan TAFIIs have been identified, yet no metazoan homologues of yeast TAFII47 (yTAFII47) and yTAFII65 are known. Both of these yTAFIIs contain a histone fold domain (HFD) which selectively heterodimerizes with that of yTAFII25. We have cloned a novel mouse protein, TAFII140, containing an HFD and a plant homeodomain (PHD) finger, which we demonstrated by immunoprecipitation to be a mammalian TFIID component. TAFII140 shows extensive sequence similarity toDrosophila BIP2 (dBIP2) (dTAFII155), which we also show to be a component of DrosophilaTFIID. These proteins are metazoan homologues of yTAFII47 as their HFDs selectively heterodimerize with dTAFII24 and human TAFII30, metazoan homologues of yTAFII25. We further show that yTAFII65 shares two domains with theDrosophila Prodos protein, a recently described potential dTAFII. These conserved domains are critical for yTAFII65 function in vivo. Our results therefore identify metazoan homologues of yTAFII47 and yTAFII65.

scholarly journals Screening For Yeast Phytase Leads to the Identification of a New Cell-Bound and Secreted Activity in Cyberlindnera jadinii CJ2

2021 ◽  
Vol 9 ◽  
Author(s):  
Claudia Capusoni ◽  
Immacolata Serra ◽  
Silvia Donzella ◽  
Concetta Compagno
Keyword(s):  
High Temperature ◽  
Phytic Acid ◽  
Convenient Method ◽  
Kluyveromyces Marxianus ◽  
Food Industry ◽  
Yeast Species ◽  
Industrial Applications ◽  
Industrial Processes ◽  
Acidic Ph ◽  
Initial Screening

Phytic acid is an anti-nutritional compound able to chelate proteins and ions. For this reason, the food industry is looking for a convenient method which allows its degradation. Phytases are a class of enzymes that catalyze the degradation of phytic acid and are used as additives in feed-related industrial processes. Due to their industrial importance, our goal was to identify new activities that exhibit best performances in terms of tolerance to high temperature and acidic pH. As a result of an initial screening on 21 yeast species, we focused our attention on phytases found in Cyberlindnera jadinii, Kluyveromyces marxianus, and Torulaspora delbrueckeii. In particular, C. jadinii showed the highest secreted and cell-bound activity, with optimum of temperature and pH at 50°C and 4.5, respectively. These characteristics suggest that this enzyme could be successfully used for feed as well as for food-related industrial applications.

scholarly journals Evaluation of top-down mass spectrometry and ion-mobility spectroscopy as a means of mapping protein-binding motifs within heparin chains

The Analyst ◽  
2020 ◽  
Vol 145 (8) ◽  
pp. 3090-3099 ◽  
Author(s):  
Yunlong Zhao ◽  
Igor A. Kaltashov
Keyword(s):  
Mass Spectrometry ◽  
Protein Binding ◽  
Ion Mobility ◽  
Structural Elements ◽  
Client Protein ◽  
Top Down ◽  
Binding Motifs ◽  
Ion Mobility Spectroscopy ◽  
High Degree ◽  
Top Down Mass Spectrometry

Identifying structural elements within glycosaminoglycans that enable their interaction with a specific client protein remains a challenging task due to the high degree of both intra- and inter-chain heterogeneity exhibited by this polysaccharide.

Novel Non-Injection Approaches for Revealing and for Microscopic Investigation of the Microcirculatory Bed in Different Organs and Tissues

1997 ◽  
Vol 3 (S2) ◽  
pp. 327-328 ◽  
Author(s):  
N. Chilingaryan ◽  
A. Chilingaryan ◽  
M. Chilingaryan
Keyword(s):  
Vascular Endothelium ◽  
Microscopic Investigation ◽  
Extensive Study ◽  
Structural Elements ◽  
Selective Precipitation ◽  
Microcirculatory Bed ◽  
Clear Cut ◽  
Injection Methods ◽  
Pathological Material ◽  
High Degree

Certain disadvantages of traditional injection methods and difficulties arising during investigation of experimental and pathological material, limit the possibilities for extensive study of the microcirculatory bed (MCB). This has prompted the authors to develop new non-injection methods for revealing the MCB on histological sections by means of direct staining of the structural elements of the vessel wall. These methods are based on selective precipitation of the extra- or intracellular orthophosphates with ions of Zn, Cd, Co, Ca, Sr, and Pb in the vascular endothelium and smooth muscle cells. These methods (which we have called “histoangiological”) provide clear-cut contrast dyeing of the MCB in different organs and tissues as well as tumors in rat, cat,pig, dog, and in humans.In these methods, the reaction end product is lead sulfide which provides a high degree of contrast. Vessels and capillaries are detected due to precipitation of the deposit in the vascular endothelium.

Role of glutamine synthetase in phenazine antibiotic production by Pantoea agglomerans Eh1087

2004 ◽  
Vol 50 (10) ◽  
pp. 877-881 ◽  
Author(s):  
Matthew D Galbraith ◽  
Stephen R Giddens ◽  
H Khris Mahanty ◽  
Bruce Clark
Keyword(s):  
Glutamine Synthetase ◽  
Antibiotic Production ◽  
Amino Acid Sequences ◽  
Pantoea Agglomerans ◽  
Wild Type ◽  
Isotopic Labelling ◽  
High Degree ◽  
Degree Of Similarity ◽  
Isotopic Labelling Experiments

Pantoea agglomerans strain Eh1087 produces the phenazine antibiotic D-alanylgriseoluteic acid. A glutamine auxotroph harboring an insertion in a putative glnA gene was obtained by transposon-mutagenesis of Eh1087 that produced less D-alanylgriseoluteic acid than the parental strain (strain Eh7.1). Cosmids encoding the Eh1087 glnA were isolated by their ability to complement the mutant for prototrophy. The role of the Eh1087 glnA locus was functionally confirmed by complementation of an Escherichia coli glnA mutant. Analysis of the nucleotide and deduced amino acid sequences of the Eh1087 glnA gene indicated a high degree of similarity to the glnA genes and glutamine synthetase enzymes of other Enterobacteriaceae. Isotopic labelling experiments with 15N-labelled ammonium sulfate demonstrated that wild-type Eh1087 incorporated 15N into griseoluteic acid more readily than the glnA mutant Eh7.1. We conclude that the 2 nitrogens in the phenazine nucleus originate from glutamine and the intracellular glutamine synthesized by Eh1087 is a source of the phenazine nucleus nitrogens even in glutamine-rich environments.Key words: phenazine, Pantoea, Erwinia, glutamine synthetase, biosynthesis.

Carnivora: The Amino Acid Sequence of the Adult European Polecat (Mustela putorius, Mustelidae) Hemoglobins

1989 ◽  
Vol 44 (7) ◽  
pp. 817-824 ◽  
Author(s):  
Aftab Ahmed ◽  
Meeno Jahan ◽  
Gerhard Braunitzer ◽  
Helmut Pechlaner
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Gas Phase ◽  
Primary Structure ◽  
Amino Acid Sequences ◽  
Mustela Putorius ◽  
Globin Chains ◽  
The Family ◽  
High Degree ◽  
Β Chain

The complete amino acid sequences of the hemoglobins from the adult European polecat (Mustela putorius) are presented. The erythrocytes contain two hemoglobin components and three globin chains (α I, α II and β). The primary structure of globin chains and of the tryptic peptides determined in liquid- and gas-phase sequantors. Comparing the sequences of the globin chains of the polecat with that of human Hb-A, 17 (23.9%) substitutions were recognized in the α I, 16 (22.5%) in the α II and 14 (20.4%) in the β chain. A high degree of homology observed with other representatives of the family Mustelidae.

Triticum aestivum L. endoxylanase inhibitor (TAXI) consists of two inhibitors, TAXI I and TAXI II, with different specificities

2001 ◽  
Vol 353 (2) ◽  
pp. 239-244 ◽  
Author(s):  
Kurt GEBRUERS ◽  
Winok DEBYSER ◽  
Hans GOESAERT ◽  
Paul PROOST ◽  
Jozef VAN DAMME ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Evolutionary Relationship ◽  
Triticum Aestivum L ◽  
Amino Acid Sequences ◽  
Molecular Structures ◽  
Molecular Forms ◽  
Inhibition Activity ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
High Degree

The Triticum aestivum L. endoxylanase inhibitor (TAXI) discovered by Debyser and Delcour [(1997) Eur. Pat. filed April 1997, published as WO 98/49278] and Debyser, Derdelinckx and Delcour [(1997) J. Am. Soc. Brew. Chem. 55, 153Ő156] seems to be a mixture of two different endoxylanase inhibitors, called TAXI I and TAXI II. By using Aspergillus niger as well as Bacillus subtilis endoxylanases for assaying inhibition activity, both inhibitors could be purified to homogeneity from wheat (Triticum aestivum L., var. Soissons). TAXI I and TAXI II have similar molecular structures. They both have a molecular mass of approx. 40.0kDa, are not glycosylated and occur in two molecular forms, i.e. a non-proteolytically processed one and a proteolytically processed one. However, the pI of TAXI II (at least 9.3) is higher than that of TAXI I (8.8). TAXI I and TAXI II clearly show different inhibition activities towards different endoxylanases. The N-terminal amino acid sequences of both inhibitors show a high degree of identity, which might indicate that there is an evolutionary relationship between them.

scholarly journals Molecular characterisation, genetic variability and detection of a functional polymorphism influencing the promoter activity of OXT gene in goat and sheep

2017 ◽  
Vol 84 (2) ◽  
pp. 165-169 ◽  
Author(s):  
Gianfranco Cosenza ◽  
Marco Iannaccone ◽  
Boipuso Alpheus Pico ◽  
Daniela Gallo ◽  
Rosanna Capparelli ◽  
...  
Keyword(s):  
Genetic Variability ◽  
Promoter Activity ◽  
Association Studies ◽  
Amino Acid Sequences ◽  
Luciferase Assay ◽  
Physiological Processes ◽  
Genetic Types ◽  
Flanking Region ◽  
Set Up ◽  
High Degree

The purpose of the study described in this Research Communication was to report the full characterisation of the goat and sheep oxytocin-neurophysin I gene (OXT), their promoters and amino acid sequences. Using the genomic DNA as template, we sequenced and compared the whole OXT gene (3 exons), plus 958/960 nucleotides at the 5′ flanking region and 478/477 nucleotides at the 3′ flanking region, in 46 sheep and 24 goats belonging to different breeds/genetic types reared in Italy, Greece and Germany. The comparison of the obtained sequences showed a high degree of genetic variability at these loci. In particular, we focused on the SNP g.438T > C as possible example of trans-specific polymorphism. This SNP alters a putative binding site of the transcription factor Oct-1. The set-up of a luciferase assay confirmed that the C variant of this SNP negatively affects the promoter activity of the sheep OXT gene. The results of this study suggest that the SNP g.438T > C might be useful to promote association studies with traits/physiological processes controlled by this hormone.

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