Isolation and characterization of a new agglutinin from the red marine alga Hypnea cervicornis J. Agardh

2006 ◽  
Vol 84 (1) ◽  
pp. 49-54 ◽  
Author(s):  
K S Nascimento ◽  
C S Nagano ◽  
E V Nunes ◽  
R F Rodrigues ◽  
G V Goersch ◽  
...  
Keyword(s):  
Disulfide Bonds ◽  
Marine Alga ◽  
Biochemical Characterization ◽  
Atlantic Coast ◽  
Haemagglutination Inhibition ◽  
Structural Diversity ◽  
Hypnea Musciformis ◽  
Isolation And Characterization ◽  
Red Marine Alga

The biochemical characterization of a new lectin (Hypnea cervicornis agglutinin or HCA) isolated from the Brazilian red alga H. cervicornis is reported. The haemagglutinating activity of the lectin was only inhibited by the glycoprotein porcine stomach mucin at a minimum inhibitory concentration of 19 µg·mL–1. No haemagglutination inhibition was detected after the addition of simple sugars. The MALDI-TOF molecular masses of native and reduced and carbamidomethylated HCA were, respectively, 9196.6 Da and 9988.2 Da, indicating that the primary structure of the protein is crosslinked by 7 disulfide bonds. This unusual structural feature among lectins, along with its N-terminal sequence and amino-acid composition, clearly shows that HCA belongs to a protein family distinct from the isolectins Hypnin A1 and A2 isolated from the related Japanese alga Hypnea japonica. On the other hand, HCA displayed a high degree of similarity to the agglutinin from the Brazilian species Hypnea musciformis. Our data indicate the occurrence of structural diversity among lectins of closely related species living in distant ecosystems, i.e., the Pacific coast of Japan and the Atlantic coast of Brazil, and support the hypothesis that the lectin content (lectinome) might serve as a biomarker for taxonomical purposes.Key words: agglutinin, lectin, isolation, Hypnea cervicornis, red marine alga.

Purification and Characterization of a new Lectin from the Red Marine Alga Hypnea Musciformis

2002 ◽  
Vol 9 (2) ◽  
pp. 159-165 ◽  
Author(s):  
C. Nagano ◽  
F.B. Moreno ◽  
C. Bloch Jr ◽  
M. Prates ◽  
J. Calvete ◽  
...  
Keyword(s):  
Marine Alga ◽  
Purification And Characterization ◽  
Hypnea Musciformis ◽  
Red Marine Alga

scholarly journals In vitro activities of kappa-carrageenan isolated from red marine alga Hypnea musciformis: Antimicrobial, anticancer and neuroprotective potential

2018 ◽  
Vol 112 ◽  
pp. 1248-1256 ◽  
Author(s):  
Ricardo Basto Souza ◽  
Annyta Fernandes Frota ◽  
Joana Silva ◽  
Celso Alves ◽  
Agnieszka Zofia Neugebauer ◽  
...  
Keyword(s):  
Marine Alga ◽  
Hypnea Musciformis ◽  
Kappa Carrageenan ◽  
Red Marine Alga

scholarly journals Isolation and Characterization of Xenobiotic Pesticide Degrading Bacterial Species in Flower Farms Around Lake Naivasha, Kenya

2018 ◽  
Vol 1 (1) ◽  
Keyword(s):  
Biochemical Characterization ◽  
Bacterial Species ◽  
Rhodococcus Erythropolis ◽  
Soil Samples ◽  
Lake Naivasha ◽  
Organophosphate Pesticide ◽  
Lower Growth ◽  
Isolation And Characterization ◽  
Cell Counts

Certain microorganisms especially bacteria and fungi are able to use xenobiotic organic compounds as their carbon and nitrogen source for metabolism. Flower farms around lake Naivasha basin uses several agrochemicals especially pesticides to control pests and improve flower production. The aim of this study was to isolate and characterize morphologically and biochemically the main bacterial species that are able to grow and tolerate the pesticide contaminated farm soils. Soil samples were collected from randomly selected five greenhouses from each five flower farms namely Crescent, Elsamere, Karuturi, Malewa and Sewage farms around Lake Naivasha basin. The collected samples were processed for bacterial isolation using the nutrient agar, mac’ Conkey agar, blood agar, Luria-Bertani and Minimum Salt Media nutrient media. The conventional methods of swabbing and streaking were used. Pure colonies of isolates organisms were identified and characterized using standard microbiological technique. Morphological, cultural and biochemical characterization of bacterial species isolated from the flower farm soil samples identified mainly Pseudomonas auriginosa, Escherichia coli, Rhodococcus erythropolis and Bacillus subtilis species. Bacterial growth in pesticide consortia was quantified by monitoring colony growth of the species in liquid culture over time. The viable cell counts were determined turbidimetrically at O.D696nm. All the isolated bacterial species were able to grow in flower farm soil contaminated with organochloride and organophosphate pesticide residues. B. subtilis recorded the highest growth at 1.77±0.07 O.D696nm in pesticide mixture consortia. There was lower growth in organochloride pesticide consortia as compared to organophosphate pesticide consortia.

scholarly journals Evaluation of the Extent of Haptoglobin Glycosylation Using Orthogonal Intact-Mass MS Approaches

2021 ◽  
Author(s):  
Yang Yang ◽  
Jake W. Pawlowski ◽  
Ian J. Carrick ◽  
Igor Kaltashov
Keyword(s):  
Disulfide Bonds ◽  
Heterogeneous Systems ◽  
Structural Diversity ◽  
Mass Range ◽  
Protein Characterization ◽  
Charge Reduction ◽  
Average Mass ◽  
Significant Difference ◽  
Intact Mass

Intact-mass measurements are becoming an increasingly popular in mass spectrometry (MS) based protein characterization, as they allow the entire complement of proteoforms to be evaluated within a relatively short time. However, applications of this approach are currently limited to systems exhibiting relatively modest degrees of structural diversity, as the high extent of heterogeneity frequently prevents straightforward MS measurements. Incorporation of limited charge reduction into electrospray ionization (ESI) MS measurements provides an elegant way to obtain meaningful information on most heterogeneous systems, yielding not only the average mass of the protein, but also the mass range populated by various proteoforms. Application of this approach to characterization of two different phenotypes of haptoglobin (1-1 and 2-1) provides evidence of a significant difference in their extent of glycosylation, with the glycan load of phenotype 2-1 being notably lighter. More detailed characterization of their glycosylation patterns is enabled by the recently introduced crosspath reactive chromatography (XP-RC) with on-line MS detection, a technique that combines chromatographic separation with in-line reduction of disulfide bonds to generate metastable haptoglobin subunits. Application of XP-RC to both haptoglobin phenotypes confirms that no modifications are present within their light chains, and provides a wealth of information on glycosylation patterns of the heavy chains. The haptoglobin 1-1 glycans are mature fully sialylated biantennary structures that exhibit high degrees of fucosylation. In contrast, phenotype 2-1 contains a significant fraction of incomplete biantennary structures and exhibit significantly lower levels of sialylation and fucosylation. The glycosylation patterns deduced from the XP-RC/MS measurements are in agreement with the conclusions of haptoglobin analysis by limited charge reduction, suggesting that the latter can be employed in situations when a fast assessment of a protein heterogeneity is needed (e.g., comparability studies of biopharmaceutical products). <br>

scholarly journals Characterization of a Highly Thermostable Alkaline Phosphatase from the Euryarchaeon Pyrococcus abyssi

2001 ◽  
Vol 67 (10) ◽  
pp. 4504-4511 ◽  
Author(s):  
Sébastien Zappa ◽  
Jean-Luc Rolland ◽  
Didier Flament ◽  
Yannick Gueguen ◽  
Joseph Boudrant ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Disulfide Bonds ◽  
Metal Ion ◽  
Divalent Cations ◽  
E Coli ◽  
Isolation And Characterization ◽  
Pyrococcus Abyssi ◽  
Increased Activity ◽  
Metal Ion Chelators

ABSTRACT This work reports the first isolation and characterization of an alkaline phosphatase (AP) from a hyperthermophilic archaeon. An AP gene from Pyrococcus abyssi, a euryarchaeon isolated from a deep-sea hydrothermal vent, was cloned and the enzyme expressed in Escherichia coli. Analysis of the sequence showed conservation of the active site and structural elements of theE. coli AP. The recombinant AP was purified and characterized. Monomeric and homodimeric active forms were detected, with a monomer molecular mass of 54 kDa. Apparent optimum pH and temperature were estimated at 11.0 and 70°C, respectively. Thus far,P. abyssi AP has been demonstrated to be the most thermostable AP, with half-lives at 100 and 105°C of 18 and 5 h, respectively. Enzyme activity was found to be dependent on divalent cations: metal ion chelators inhibited activity, whereas the addition of exogenous Mg(II), Zn(II), and Co(II) increased activity. The enzyme was inhibited by inorganic phosphate, but not by molybdate and vanadate. Strong inhibitory effects were observed in the presence of thiol-reducing agents, although cysteine residues of the P. abyssi AP were not found to be incorporated within intra- or interchain disulfide bonds. In addition,P. abyssi AP was demonstrated to dephosphorylate linear DNA fragments with dephosphorylation efficiencies of 93.8 and 84.1% with regard to cohesive and blunt ends, respectively.

scholarly journals Isolation and characterization of two O-methyltransferases involved in benzylisoquinoline alkaloid biosynthesis in sacred lotus (Nelumbo nucifera)

2019 ◽  
Vol 295 (6) ◽  
pp. 1598-1612 ◽  
Author(s):  
Ivette M. Menéndez-Perdomo ◽  
Peter J. Facchini
Keyword(s):  
Biochemical Characterization ◽  
Functional Characterization ◽  
Nelumbo Nucifera ◽  
Plant Metabolites ◽  
Sacred Lotus ◽  
Isolation And Characterization ◽  
Medicinal Value ◽  
Benzylisoquinoline Alkaloid ◽  
Young Leaves

Benzylisoquinoline alkaloids (BIAs) are a major class of plant metabolites with many pharmacological benefits. Sacred lotus (Nelumbo nucifera) is an ancient aquatic plant of medicinal value because of antiviral and immunomodulatory activities linked to its constituent BIAs. Although more than 30 BIAs belonging to the 1-benzylisoquinoline, aporphine, and bisbenzylisoquinoline structural subclasses and displaying a predominant R-enantiomeric conformation have been isolated from N. nucifera, its BIA biosynthetic genes and enzymes remain unknown. Herein, we report the isolation and biochemical characterization of two O-methyltransferases (OMTs) involved in BIA biosynthesis in sacred lotus. Five homologous genes, designated NnOMT1–5 and encoding polypeptides sharing >40% amino acid sequence identity, were expressed in Escherichia coli. Functional characterization of the purified recombinant proteins revealed that NnOMT1 is a regiospecific 1-benzylisoquinoline 6-O-methyltransferase (6OMT) accepting both R- and S-substrates, whereas NnOMT5 is mainly a 7-O-methyltransferase (7OMT), with relatively minor 6OMT activity and a strong stereospecific preference for S-enantiomers. Available aporphines were not accepted as substrates by either enzyme, suggesting that O-methylation precedes BIA formation from 1-benzylisoquinoline intermediates. Km values for NnOMT1 and NnOMT5 were 20 and 13 μm for (R,S)-norcoclaurine and (S)-N-methylcoclaurine, respectively, similar to those for OMTs from other BIA-producing plants. Organ-based correlations of alkaloid content, OMT activity in crude extracts, and OMT gene expression supported physiological roles for NnOMT1 and NnOMT5 in BIA metabolism, occurring primarily in young leaves and embryos of sacred lotus. In summary, our work identifies two OMTs involved in BIA metabolism in the medicinal plant N. nucifera.

Structure of mouse muskelin discoidin domain and biochemical characterization of its self-association

2014 ◽  
Vol 70 (11) ◽  
pp. 2863-2874 ◽  
Author(s):  
Kook-Han Kim ◽  
Seung Kon Hong ◽  
Kwang Yeon Hwang ◽  
Eunice EunKyeong Kim
Keyword(s):  
Crystal Structure ◽  
Cell Adhesion ◽  
Disulfide Bonds ◽  
Biochemical Characterization ◽  
Specific Interaction ◽  
Repeat Domain ◽  
Interdomain Interaction ◽  
Self Association ◽  
Cytoskeleton Dynamics

Muskelin is an intracellular kelch-repeat protein comprised of discoidin, LisH, CTLH and kelch-repeat domains. It is involved in cell adhesion and the regulation of cytoskeleton dynamics as well as being a component of a putative E3 ligase complex. Here, the first crystal structure of mouse muskelin discoidin domain (MK-DD) is reported at 1.55 Å resolution, which reveals a distorted eight-stranded β-barrel with two short α-helices at one end of the barrel. Interestingly, the N- and C-termini are not linked by the disulfide bonds found in other eukaryotic discoidin structures. A highly conserved MIND motif appears to be the determinant for MK-DD specific interaction together with the spike loops. Analysis of interdomain interaction shows that MK-DD binds the kelch-repeat domain directly and that this interaction depends on the presence of the LisH domain.

scholarly journals Isolation and Characterization of Brenneria quercina, Causal Agent for Bark Canker and Drippy Nut of Quercus spp. in Spain

2003 ◽  
Vol 93 (4) ◽  
pp. 485-492 ◽  
Author(s):  
Elena G. Biosca ◽  
Raquel González ◽  
María José López-López ◽  
Santiago Soria ◽  
Carmina Montón ◽  
...  
Keyword(s):  
Fungal Pathogens ◽  
Reference Strain ◽  
Biochemical Characterization ◽  
Causal Agent ◽  
Enzyme Linked Immunosorbent Assay ◽  
Isolation And Characterization ◽  
Pathogenicity Tests ◽  
Different Origins ◽  
First Time

The drippy nut disease of oak was first described in California in 1967 and, since then, the causal agent has not been reported in any other area. This study describes for the first time in Europe the isolation of Brenneria (Erwinia) quercina from bark canker in addition to drippy bud and drippy nut in Quercus ilex and Q. pyrenaica. The bark canker and drippy bud symptoms were not previously described as caused by this bacterium. No fungal pathogens were associated with any of the symptoms. Physiological and biochemical characterization identified the pathogenic isolates from Spain as belonging to B. quercina, similar to the reference strain CFBP 1266. Fatty acid profiles of the Spanish isolates also were similar to the strain of B. quercina from California. Serological analysis by indirect immunofluorescence and enzyme-linked immunosorbent assay using polyclonal antisera against the reference strain of B. quercina and one Spanish oak isolate revealed some antigenic heterogeneity between isolates of different origins. Pathogenicity tests demonstrated that the Spanish isolates were able to reproduce internal symptoms of necrosis and acorn exudation in Q. ilex and Q. pyrenaica and suggest that B. quercina may be associated, among other causes, with the oak decline syndrome affecting Spanish oak forests.

scholarly journals Membrane retrieval in the guinea-pig neurohypophysis. Isolation and characterization of secretory vesicles and coated microvesicles after radiolabel incorporation in vivo

1984 ◽  
Vol 218 (2) ◽  
pp. 591-599 ◽  
Author(s):  
T Saermark ◽  
P M Jones ◽  
I C A F Robinson
Keyword(s):  
Guinea Pig ◽  
Biochemical Characterization ◽  
Hormone Secretion ◽  
Secretory Vesicle ◽  
Small Scale ◽  
Secretory Vesicles ◽  
Isolation And Characterization ◽  
Pituitary Glands

We have developed small-scale methods for the isolation and biochemical characterization of subcellular fractions from single guinea-pig posterior-pituitary glands. Secretory vesicles and coated microvesicles produced in this way were of similar purity to those isolated from large amounts of tissue by conventional ultracentrifugation. [35S]Cysteine injected into the hypothalamus was found in the soluble contents of secretory vesicles isolated from the neural lobes 24 h later. High-pressure liquid-chromatographic analysis revealed that the radiolabel was incorporated into the expected neurosecretory products (oxytocin, vasopressin and neurophysin) and also into a biosynthetic intermediate in the vasopressin system. The membranes of secretory vesicles were labelled with [3H]choline 24 h after its hypothalamic injection. Little or no [3H]choline could be demonstrated in coated microvesicles at this time, although these structures were labelled 5 days after injection. Stimulating hormone secretion by chronic dehydration produced a significant fall in [3H]choline content of the secretory-vesicle membranes without any transfer of label into coated microvesicles, suggesting that coated microvesicles are not involved in membrane retrieval in the neurohypophysis.

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