scholarly journals Profile of Secreted Hydrolases, Associated Proteins, and SlpA in Thermoanaerobacterium saccharolyticum during the Degradation of Hemicellulose

2014 ◽  
Vol 80 (16) ◽  
pp. 5001-5011 ◽  
Author(s):  
D. H. Currie ◽  
A. M. Guss ◽  
C. D. Herring ◽  
R. J. Giannone ◽  
C. M. Johnson ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Transmembrane Proteins ◽  
Liquid Chromatography Mass Spectrometry ◽  
Phosphotransferase System ◽  
Content Type ◽  
Thermoanaerobacterium Saccharolyticum ◽  
Conditional Expression ◽  
Hemicellulose Degradation ◽  
Associated Proteins

ABSTRACTThermoanaerobacterium saccharolyticum, a Gram-positive thermophilic anaerobic bacterium, grows robustly on insoluble hemicellulose, which requires a specialized suite of secreted and transmembrane proteins. We report here the characterization of proteins secreted by this organism. Cultures were grown on hemicellulose, glucose, xylose, starch, and xylan in pH-controlled bioreactors, and samples were analyzed via spotted microarrays and liquid chromatography-mass spectrometry. Key hydrolases and transporters employed byT. saccharolyticumfor growth on hemicellulose were, for the most part, hitherto uncharacterized and existed in two clusters (Tsac_1445throughTsac_1464for xylan/xylose andTsac_1344throughTsac_1349for starch). A phosphotransferase system subunit, Tsac_0032, also appeared to be exclusive to growth on glucose. Previously identified hydrolases that showed strong conditional expression changes included XynA (Tsac_1459), XynC (Tsac_0897), and a pullulanase, Apu (Tsac_1342). An omnipresent transcript and protein making up a large percentage of the overall secretome, Tsac_0361, was tentatively identified as the primary S-layer component inT. saccharolyticum, and deletion of theTsac_0361gene resulted in gross morphological changes to the cells. The view of hemicellulose degradation revealed here will be enabling for metabolic engineering efforts in biofuel-producing organisms that degrade cellulose well but lack the ability to catabolize C5sugars.

Microscopic characterization of acidic paper manuscripts

2019 ◽  
Vol 48 (2) ◽  
pp. 119-128 ◽  
Author(s):  
Safa Abd El kader Mohamed Hamed ◽  
Rushdya Rabee Ali Hassan
Keyword(s):  
Ph Value ◽  
Morphological Changes ◽  
Environmental Scanning ◽  
X Ray Diffraction ◽  
Arabic Gum ◽  
Content Type ◽  
Cellulosic Fibers ◽  
Fiber Degradation ◽  
Historical Samples

Purpose This paper aims to investigate the effect of acidity on the morphology of archeological paper, especially in the presence of colors and whether natural pigments play a role in the process of degradation. Design/methodology/approach The morphological changes in the cellulosic fibers of the manuscripts because of acidity were investigated using environmental scanning electron microscope (ESEM). Ten historical samples were collected from different manuscripts suffering from acidity. X-ray diffraction was used to identify the inks and pigments that were used in some samples. Additionally, Fourier transform infrared microscopy was used to identify the binding medium. Findings The results confirmed that carbon ink, ultramarine, cinnabar and gold pigments were applied to some manuscripts with Arabic gum. As for ESEM investigation, the results proved that acidity badly affected the integrity of the cellulosic fibers resulting in their embrittlement. The micrographs showed differences in fiber degradation according to pH value. The presence of inks and pigments increased the degradation extent resulting from acidity. Originality/value This paper addresses a specific need to study the behavior of degradation in paper manuscripts, thus helping the conservators find solutions to the phenomenon.

scholarly journals Identification and Characterization of the Rhoptry Neck Protein 2 in Babesia divergens and B. microti

2016 ◽  
Vol 84 (5) ◽  
pp. 1574-1584 ◽  
Author(s):  
Rosalynn L. Ord ◽  
Marilis Rodriguez ◽  
Jeny R. Cursino-Santos ◽  
Hyunryung Hong ◽  
Manpreet Singh ◽  
...  
Keyword(s):  
Direct Role ◽  
Parasite Invasion ◽  
Content Type ◽  
Apicomplexan Parasites ◽  
Babesia Divergens ◽  
Associated Proteins ◽  
Identification And Characterization

Apicomplexan parasites include those of the generaPlasmodium,Cryptosporidium, andToxoplasmaand those of the relatively understudied zoonotic genusBabesia. In humans, babesiosis, particularly transfusion-transmitted babesiosis, has been emerging as a major threat to public health. Like malaria, the disease pathology is a consequence of the parasitemia which develops through cyclical replication ofBabesiaparasites in host erythrocytes. However, there are no exoerythrocytic stages inBabesia, so targeting of the blood stage and associated proteins to directly prevent parasite invasion is the most desirable option for effective disease control. Especially promising among such molecules are the rhoptry neck proteins (RONs), whose homologs have been identified in many apicomplexan parasites. RONs are involved in the formation of the moving junction, along with AMA1, but no RON has been identified and characterized in anyBabesiaspp. Here we identify the RON2 proteins ofBabesia divergens(BdRON2) andB. microti(BmRON2) and show that they are localized apically and that anti-BdRON2 antibodies are significant inhibitors of parasite invasionin vitro. Neither protein is immunodominant, as both proteins react only marginally with sera from infected animals. Further characterization of the direct role of both BdRON2 and BmRON2 in parasite invasion is required, but knowledge of the level of conformity of RON2 proteins within the apicomplexan phylum, particularly that of the AMA1-RON2 complex at the moving junction, along with the availability of an animal model forB. microtistudies, provides a key to target this complex with a goal of preventing the erythrocytic invasion of these parasites and to further our understanding of the role of these conserved ligands in invasion.

scholarly journals Characterization of Xylan Utilization and Discovery of a New Endoxylanase in Thermoanaerobacterium saccharolyticum through Targeted Gene Deletions

2012 ◽  
Vol 78 (23) ◽  
pp. 8441-8447 ◽  
Author(s):  
Kara K. Podkaminer ◽  
Adam M. Guss ◽  
Heather L. Trajano ◽  
David A. Hogsett ◽  
Lee R. Lynd
Keyword(s):  
Cellulose Hydrolysis ◽  
High Yield ◽  
Glycoside Hydrolase Family ◽  
Xylose Utilization ◽  
Gene Deletions ◽  
Content Type ◽  
Xylan Hydrolysis ◽  
Thermoanaerobacterium Saccharolyticum ◽  
Single Deletion

ABSTRACTThe economical production of fuels and commodity chemicals from lignocellulose requires the utilization of both the cellulose and hemicellulose fractions. Xylanase enzymes allow greater utilization of hemicellulose while also increasing cellulose hydrolysis. Recent metabolic engineering efforts have resulted in a strain ofThermoanaerobacterium saccharolyticumthat can convert C5and C6sugars, as well as insoluble xylan, into ethanol at high yield. To better understand the process of xylan solubilization in this organism, a series of targeted deletions were constructed in the homoethanologenicT. saccharolyticumstrain M0355 to characterize xylan hydrolysis and xylose utilization in this organism. While the deletion of β-xylosidasexylDslowed the growth ofT. saccharolyticumon birchwood xylan and led to an accumulation of short-chain xylo-oligomers, no other single deletion, including the deletion of the previously characterized endoxylanase XynA, had a phenotype distinct from that of the wild type. This result indicates a multiplicity of xylanase enzymes which facilitate xylan degradation inT. saccharolyticum. Growth on xylan was prevented only when a previously uncharacterized endoxylanase encoded byxynCwas also deleted in conjunction withxynA. Sequence analysis ofxynCindicates that this enzyme, a low-molecular-weight endoxylanase with homology to glycoside hydrolase family 11 enzymes, is secreted yet untethered to the cell wall. Together, these observations expand our understanding of the enzymatic basis of xylan hydrolysis byT. saccharolyticum.

scholarly journals New Aspects of the Interplay between Penicillin Binding Proteins, murM, and the Two-Component System CiaRH of Penicillin-Resistant Streptococcus pneumoniae Serotype 19A Isolates from Hungary

2017 ◽  
Vol 61 (7) ◽  
Author(s):  
Inga Schweizer ◽  
Sebastian Blättner ◽  
Patrick Maurer ◽  
Katharina Peters ◽  
Daniela Vollmer ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Morphological Changes ◽  
Penicillin Resistance ◽  
Penicillin Binding Protein ◽  
Two Component System ◽  
Gene Encoding ◽  
Content Type ◽  
Penicillin Binding Proteins ◽  
High Level

ABSTRACT The Streptococcus pneumoniae clone Hungary19A-6 expresses unusually high levels of β-lactam resistance, which is in part due to mutations in the MurM gene, encoding a transferase involved in the synthesis of branched peptidoglycan. Moreover, it contains the allele ciaH232, encoding the histidine kinase CiaH (M. Müller, P. Marx, R. Hakenbeck, and R. Brückner, Microbiology 157:3104–3112, 2011, https://doi.org/10.1099/mic.0.053157-0 ). High-level penicillin resistance primarily requires the presence of low-affinity (mosaic) penicillin binding protein (PBP) genes, as, for example, in strain Hu17, a closely related member of the Hungary19A-6 lineage. Interestingly, strain Hu15 is β-lactam sensitive due to the absence of mosaic PBPs. This unique situation prompted us to investigate the development of cefotaxime resistance in transformation experiments with genes known to play a role in this phenotype, pbp2x, pbp1a, murM, and ciaH, and penicillin-sensitive recipient strains R6 and Hu15. Characterization of phenotypes, peptidoglycan composition, and CiaR-mediated gene expression revealed several novel aspects of penicillin resistance. The murM gene of strain Hu17 (murM Hu17), which is highly similar to murM of Streptococcus mitis, induced morphological changes which were partly reversed by ciaH232. murM Hu17 conferred cefotaxime resistance only in the presence of the pbp2x of strain Hu17 (pbp2x Hu17). The ciaH232 allele contributed to a remarkable increase in cefotaxime resistance in combination with pbp2x Hu17 and pbp1a of strain Hu17 (pbp1a Hu17), accompanied by higher levels of expression of CiaR-regulated genes, documenting that ciaH232 responds to PBP1aHu17-mediated changes in cell wall synthesis. Most importantly, the proportion of branched peptides relative to the proportion of linear muropeptides increased in cells containing mosaic PBPs, suggesting an altered enzymatic activity of these proteins.

scholarly journals Identification and characterization of two transmembrane proteins required for virulence of Ustilago maydis

2021 ◽  
Author(s):  
Paul Weiland ◽  
Florian Altegoer
Keyword(s):  
Virulence Factors ◽  
Biochemical Analysis ◽  
Transmembrane Proteins ◽  
Ustilago Maydis ◽  
Effector Proteins ◽  
Smut Fungi ◽  
Initial Characterization ◽  
Associated Proteins ◽  
Identification And Characterization

AbstractSmut fungi comprise a large group of biotrophic phytopathogens infecting important crops such as wheat and corn. Through the secretion of effector proteins, the fungus actively suppresses plant immune reactions and modulates its host’s metabolism. Consequently, how soluble effector proteins contribute to virulence is already characterized in a range of phytopathogens. However, membrane-associated virulence factors have been much less studied to date. Here, we investigated six transmembrane (TM) proteins that show elevated gene expression during biotrophic development of the maize pathogen Ustilago maydis. We show that two of the six proteins, named Vmp1 and Vmp2 (virulence-associated membrane protein), are essential for the full virulence of U. maydis. The deletion of the corresponding genes lead to a substantial attenuation in the virulence of U. maydis. Furthermore, both are conserved in various related smuts and contain no domains of known function. Our biochemical analysis clearly shows that Vmp1 and Vmp2 are membrane-associated proteins, potentially localizing to the U. maydis plasma membrane. Mass photometry and light scattering suggest that Vmp1 mainly occurs as a monomer, while Vmp2 is dimeric. Notably, the large and partially unstructured C-terminal domain of Vmp2 is crucial for virulence while not contributing to dimerization. Taken together, we here provide an initial characterization of two membrane proteins as virulence factors of U. maydis.

scholarly journals Identification and Characterization of Two Transmembrane Proteins Required for Virulence of Ustilago maydis

2021 ◽  
Vol 12 ◽  
Author(s):  
Paul Weiland ◽  
Florian Altegoer
Keyword(s):  
Virulence Factors ◽  
Biochemical Analysis ◽  
Transmembrane Proteins ◽  
Ustilago Maydis ◽  
Effector Proteins ◽  
Smut Fungi ◽  
Initial Characterization ◽  
Associated Proteins ◽  
Identification And Characterization

Smut fungi comprise a large group of biotrophic phytopathogens infecting important crops such as wheat and corn. Through the secretion of effector proteins, the fungus actively suppresses plant immune reactions and modulates its host’s metabolism. Consequently, how soluble effector proteins contribute to virulence is already characterized in a range of phytopathogens. However, membrane-associated virulence factors have been much less studied to date. Here, we investigated six transmembrane (TM) proteins that show elevated gene expression during biotrophic development of the maize pathogen Ustilago maydis. We show that two of the six proteins, named Vmp1 and Vmp2 (virulence-associated membrane protein), are essential for the full virulence of U. maydis. The deletion of the corresponding genes leads to a substantial attenuation in the virulence of U. maydis. Furthermore, both are conserved in various related smuts and contain no domains of known function. Our biochemical analysis clearly shows that Vmp1 and Vmp2 are membrane-associated proteins, potentially localizing to the U. maydis plasma membrane. Mass photometry and light scattering suggest that Vmp1 mainly occurs as a monomer, while Vmp2 is dimeric. Notably, the large and partially unstructured C-terminal domain of Vmp2 is crucial for virulence while not contributing to dimerization. Taken together, we here provide an initial characterization of two membrane proteins as virulence factors of U. maydis.

Thermal characterization of ABS-Graphene blended three dimensional printed functional prototypes for electro chemical energy storage

2018 ◽  
Vol 1 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Kamaljit Singh Boparai ◽  
Rupinder Singh
Keyword(s):  
Energy Storage ◽  
Structural Changes ◽  
Fused Deposition Modeling ◽  
Morphological Changes ◽  
Three Dimensional ◽  
Thermal Characterization ◽  
Chemical Energy ◽  
Flow Index ◽  
Fused Deposition

This study highlights the thermal characterization of ABS-Graphene blended three dimensional (3D) printed functional prototypes by fused deposition modeling (FDM) process. These functional prototypes have some applications as electro-chemical energy storage devices (EESD). Initially, the suitability of ABS-Graphene composite material for FDM applications has been examined by melt flow index (MFI) test. After establishing MFI, the feedstock filament for FDM has been prepared by an extrusion process. The fabricated filament has been used for printing 3D functional prototypes for printing of in-house EESD. The differential scanning calorimeter (DSC) analysis was conducted to understand the effect on glass transition temperature with the inclusion of Graphene (Gr) particles. It has been observed that the reinforced Gr particles act as a thermal reservoir (sink) and enhances its thermal/electrical conductivity. Also, FT-IR spectra realized the structural changes with the inclusion of Gr in ABS matrix. The results are supported by scanning electron microscopy (SEM) based micrographs for understanding the morphological changes.

scholarly journals Use of Proteins Identified through a Functional Genomic Screen To Develop a Protein Subunit Vaccine That Provides Significant Protection against Virulent Streptococcus suis in Pigs

2017 ◽  
Vol 86 (3) ◽  
Author(s):  
Susan L. Brockmeier ◽  
Crystal L. Loving ◽  
Tracy L. Nicholson ◽  
Jinhong Wang ◽  
Sarah E. Peters ◽  
...  
Keyword(s):  
Immune Response ◽  
Vaccine Trial ◽  
Streptococcus Suis ◽  
Protein Subunit ◽  
Content Type ◽  
Degree Of Protection ◽  
Wide Range ◽  
Associated Proteins ◽  
Clinical Protection ◽  
Cellular Immune

ABSTRACT Streptococcus suis is a bacterium that is commonly carried in the respiratory tract and that is also one of the most important invasive pathogens of swine, commonly causing meningitis, arthritis, and septicemia. Due to the existence of many serotypes and a wide range of immune evasion capabilities, efficacious vaccines are not readily available. The selection of S. suis protein candidates for inclusion in a vaccine was accomplished by identifying fitness genes through a functional genomics screen and selecting conserved predicted surface-associated proteins. Five candidate proteins were selected for evaluation in a vaccine trial and administered both intranasally and intramuscularly with one of two different adjuvant formulations. Clinical protection was evaluated by subsequent intranasal challenge with virulent S. suis . While subunit vaccination with the S. suis proteins induced IgG antibodies to each individual protein and a cellular immune response to the pool of proteins and provided substantial protection from challenge with virulent S. suis , the immune response elicited and the degree of protection were dependent on the parenteral adjuvant given. Subunit vaccination induced IgG reactive against different S. suis serotypes, indicating a potential for cross protection.

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