scholarly journals Biochemical characterisation of chlorophyllase from leaves of selected Prunus species--a comparative study.

2013 ◽  
Vol 60 (3) ◽  
Author(s):  
Hubert Sytykiewicz ◽  
Iwona Sprawka ◽  
Paweł Czerniewicz ◽  
Cezary Sempruch ◽  
Bogumił Leszczyński ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Sour Cherry ◽  
Inhibitory Effect ◽  
Biochemical Properties ◽  
Prunus Domestica ◽  
Prunus Species ◽  
Chl A ◽  
Chlorophyll Breakdown ◽  
Biochemical Characterisation ◽  
Catabolic Process

Despite senescence-induced chlorophyll depletion in plants has been widely studied, the enzymatic background of this physiologically regulated process still remains highly unclear. The purpose of this study was to determine selected biochemical properties of partially purified fractions of chlorophyllase (Chlase, chlorophyll chlorophyllido-hydrolase, EC 3.1.1.14) from leaves of three Prunus species: bird cherry (Prunus padus L.), European plum (Prunus domestica L.), and sour cherry (Prunus cerasus L.). Secondarily, this report was aimed at comparing seasonal dynamics of Chlase activity and chlorophyll a (Chl a) content within investigated plant systems. Molecular weight of native Chlase F1 has been estimated at 90 kDa (bird cherry) and approximately 100 kDa (European plum and sour cherry), whereas molecular mass of Chlase F2 varied from 35 kDa (European plum) to 60 kDa (sour cherry). Furthermore, enzyme fractions possessed similar optimal pH values ranging from 7.6 to 8.0. It was found that among a broad panel of tested metal ions, Hg(+2), Fe(+2), and Cu(+2) cations showed the most pronounced inhibitory effect on the activity of Chlase. In contrast, the presence of Mg(+2) ions influenced a subtle stimulation of the enzymatic activity. Importantly, although Chlase activity was negatively correlated with the amount of Chl a in leaves of examined Prunus species, detailed comparative analyses revealed an incidental decrement of enzymatic activity in early or moderately senescing leaves. It provides evidence that foliar Chlase is not the only enzyme involved in autumnal chlorophyll breakdown and further in-depth studies elucidating this catabolic process are required.

scholarly journals Biochemical characterisation of α-glucosidase and β-glucosidase in the alimentary canal of larval Leptinotarsa decemlineata SAY, 1824 (Coleoptera: Chrysomelidae)

2014 ◽  
Vol 83 (4) ◽  
pp. 281-294 ◽  
Author(s):  
Majid Kazzazi ◽  
Fahimeh Dehghanikhah ◽  
Hossein Madadi ◽  
Vahid Hossseininaveh
Keyword(s):  
Incubation Time ◽  
Leptinotarsa Decemlineata ◽  
Digestive Enzymes ◽  
Specific Activity ◽  
Inhibitory Effect ◽  
Maximum Activity ◽  
Pest Population ◽  
Biochemical Characterisation ◽  
Leptinotarsa Decemlineata Say ◽  
Environmentally Safe

ABSTRACT Host plant resistance is an environmentally safe method used for reducing a pest population. Basically, when developing resistant cultivars one needs to study the biochemical characteristics of the digestive enzymes in the insect’s midgut. In this study, the activities of α- and β-glucosidase were determined from Leptinotarsa decemlineata midgut using p-nitrophenyl-α-Dglucopyranoside and p-nitrophenyl-β-D-glucopyranoside as substrates respectively. The results showed that the specific activity of α- and β-glucosidase from 4th instar larvae midguts of L. decemlineata were 5.14 and 5.48 Umg-1 protein respectively. The activity of α-glucosidase was optimal at pH 4, whereas the maximum activity of β-glucosidase in the midgut of L. decemlineata occurred at pH 4-5.5. Both enzymes were stable at pH 3-8 over an incubation time of 8 hours. The respective activities of α- and β-glucosidase were at their highest at 45 °C and 50 °C, but they were not stable at 50 °C during an incubation time of 8 days. Furthermore, our data showed that MgCl2, Tris and urea have a moderate but SDS a severe inhibitory effect on enzyme activity. Biochemical characterisation revealed one and three bands of α- and β-glucosidase activities in the midgut of L. decemlineata respectively.

scholarly journals First Report of Apricot vein clearing-associated virus (AVCaV) infecting Prunus domestica revealed by High-Throughput Sequencing in Morocco

Plant Disease ◽  
2020 ◽  
Author(s):  
Rachid Tahzima ◽  
Radouane Qessaoui ◽  
Yoika Foucart ◽  
Sebastian Massart ◽  
Kris De Jonghe
Keyword(s):  
Fruit Trees ◽  
Plant Viruses ◽  
Amino Acid Sequences ◽  
Stone Fruit ◽  
Replicase Gene ◽  
Prunus Domestica ◽  
Prunus Species ◽  
First Report ◽  
Vein Clearing ◽  
The Impact

Plum (Prunus domestica L., Rosaceae) trees, like many stone fruit trees, are known to be infected by numerous plant viruses, predominantly as consequence of their clonal mode of propagation and perennial cultivation (Jelkmann and Eastwell, 2011). Apricot vein clearing-associated virus (AVCaV) is a member of the genus Prunevirus in the family Betaflexiviridae. AVCaV was first reported in Italy infecting apricot (P. armeniaca L.) associated with foliar vein clearing symptoms (Elbeaino et al. 2014). It has also been detected in various Prunus species, like plum, Japanese plum (P. salicina L.), sour cherry (P. cerasus L.), and Japanese apricot (P. mume L.), apricot and peach (P. persica L.) sourced from Asian and European countries (Marais et al. 2015), as well as in the ornamental Myrobolan plum (P. cerasifera L.) in Australia (Kinoti et al. 2017). In 2018, during the vegetative season, a survey was carried out in two different apricot and plum orchards in the southern region of Agdez (Agadir, Morocco) where stone fruit trees are grown. Five branches with leaves were sampled from three apricot and three plum trees of unknown cultivars, all asymptomatic. Total RNA was extracted from 100 mg plant tissue (leaves and cambial scrapping) using RNeasy Plant Mini Kit (QIAGEN, Hilden, Germany) and separate samples (one per species) were used for library preparation (NEBNext Ultra RNA library kit; New England BioLabs, MA, USA), and sequencing (Illumina NextSeq v2, totRNA sequencing) at Admera Health (New Jersey, USA). All generated reads (6,756,881) from the plum sample were quality filtered and submitted to the VirusDetect pipeline (Zheng et al., 2017). The plum cDNA library, a total of 20 viral contigs (68-1928 bp) mapped to several AVCaV accessions in GenBank. A reference mapping (CLC Genomics Workbench 12, Qiagen, Denmark) was conducted against all four available AVCaV full genomes (KM507062-63, KY132099 and HG008921), revealing 100% coverage of the full sequence (8358 nt) with 97-98 % nucleotide (nt) identities (BLASTn). Analysis of the derived sequences allowed to identify the location of the four predicted ORFs i.e. (ORF1: 6066 nt/2,021 aa), (ORF2: 1383 nt/460 aa), (ORF3: 666 nt/221 aa) and (ORF4: 420 nt/139 aa), previously described for the AVCaV genome (Elbeaino et al. 2014). The amino acid sequences of the encoded proteins of AVCaV isolate from Morocco also shared 97-98% identities with the corresponding sequences of complete genome AVCaV isolates in GenBank. To confirm the detection of AVCaV in the three plum samples, specific RT-PCR primers (VC37657s: 5’-CCATAGCCACCCTTTTTCAA-3’ / VC28239a: 5’-GTCGTCAAGGGTCCAGTGAT-3’) (Elbeaino et al. 2014) were used and the expected 330 bp fragment from the replicase gene was amplified in all three samples and subsequently sequenced (MT980794-96). Sanger sequences were 100% identical to corresponding HTS derived sequence. This is the first report of AVCaV infecting plum in Africa. The incidence of AVCaV in Moroccan Prunus species is unknown. Plum trees from the surveyed orchards were also confirmed to be co-infected with little cherry virus 1 (LChV-1) using HTS. Further investigation is required to determine the impact of AVCaV on these asymptomatic plum trees and other stone fruits species.

scholarly journals ENHANCED ENZYMATIC ACTIVITY OF STREPTOMYCES GRISEOPLANUS L-ASPARGINASE VIA ITS INCORPORATION IN AN OIL-BASED NANOCARRIER

2020 ◽  
pp. 203-210
Author(s):  
ABEER A. EL-HADI ◽  
HANAN MOSTAFA AHMED ◽  
RANIA A. ZAKI ◽  
AMIRA MOHAMED MOHSEN
Keyword(s):  
Thermal Stability ◽  
Oleic Acid ◽  
Enzymatic Activity ◽  
Specific Activity ◽  
Residual Activity ◽  
Tween 80 ◽  
Biochemical Properties ◽  
Lymphocytic Leukemia ◽  
Tween 20 ◽  
Ph Range

Objective: L-asparaginase (L-asp) is a vital enzyme used as a therapeutic agent in combination with other drugs in the treatment of acute lymphoma, melanosarcoma and lymphocytic leukemia. Immobilization of enzymes through loading on nanoemulsion (NE) results in some advantages such as enhancing their stability and increasing their resistance to proteases. Aim of the present study is to formulate L-asp loaded nanoemulsion to enhance its efficiency and thermal stability. Methods: Nanoemulsion loaded with L-asp crude extract (specific activity 13.23U/mg protein) was prepared employing oleic acid as oil, tween 20/tween 80 as surfactants and propylene glycol (PG) as co-surfactant. L-asp loaded NE underwent several thermodynamic stability studies and the optimized formulae were further examined for their biochemical properties and thermal stability. Results The developed formulations were spherical in shape and their sizes were in the nanometric dimensions with negatively charged zeta potential values. Upon comparing the enzyme activity of L-asp loaded NE employing tween 20 (F1) or tween80 (F4) at different concentrations, the results revealed that F4 NE showed higher enzymatic activity [323 U/ml] compared to F1 NE [197 U/ml] at the same concentration. The nanosized immobilized L-asp was more stable in the pH range from 8 to 8.5 as compared to free L-asp. The immobilized enzyme preserved about 59.11% of its residual activity at 50 °C; while free L-asp preserved about 33.84%. Conclusion: In the view of these results, NE composed of oleic acid, tween 80 and PG represents a promising dosage form for enhancing the activity and stability of Streptomyces griseoplanus L-asp.

scholarly journals The Pharmacological Activity, Biochemical Properties, and Pharmacokinetics of the Major Natural Polyphenolic Flavonoid: Quercetin

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 374 ◽  
Author(s):  
Gaber El-Saber Batiha ◽  
Amany Magdy Beshbishy ◽  
Muhammad Ikram ◽  
Zohair S. Mulla ◽  
Mohamed E. Abd El-Hack ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
Biological Activities ◽  
Inhibitory Effect ◽  
Biochemical Properties ◽  
Beneficial Effects ◽  
Natural Substances ◽  
Current Review ◽  
Flavonoid Quercetin ◽  
Small Intestines ◽  
The Brain

Flavonoids are a class of natural substances present in plants, fruits, vegetables, wine, bulbs, bark, stems, roots, and tea. Several attempts are being made to isolate such natural products, which are popular for their health benefits. Flavonoids are now seen as an essential component in a number of cosmetic, pharmaceutical, and medicinal formulations. Quercetin is the major polyphenolic flavonoid found in food products, including berries, apples, cauliflower, tea, cabbage, nuts, and onions that have traditionally been treated as anticancer and antiviral, and used for the treatment of allergic, metabolic, and inflammatory disorders, eye and cardiovascular diseases, and arthritis. Pharmacologically, quercetin has been examined against various microorganisms and parasites, including pathogenic bacteria, viruses, and Plasmodium, Babesia, and Theileria parasites. Additionally, it has shown beneficial effects against Alzheimer’s disease (AD), and this activity is due to its inhibitory effect against acetylcholinesterase. It has also been documented to possess antioxidant, antifungal, anti-carcinogenic, hepatoprotective, and cytotoxic activity. Quercetin has been documented to accumulate in the lungs, liver, kidneys, and small intestines, with lower levels seen in the brain, heart, and spleen, and it is extracted through the renal, fecal, and respiratory systems. The current review examines the pharmacokinetics, as well as the toxic and biological activities of quercetin.

scholarly journals Soil Resistance to Burn Severity in Different Forest Ecosystems in the Framework of a Wildfire

Forests ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 773
Author(s):  
Sara Huerta ◽  
Víctor Fernández-García ◽  
Leonor Calvo ◽  
Elena Marcos
Keyword(s):  
Enzymatic Activity ◽  
Forest Ecosystems ◽  
Resistance To Change ◽  
Chemical Properties ◽  
Resistance Index ◽  
Biochemical Properties ◽  
Burn Severity ◽  
Total N ◽  
Organic C ◽  
Soil Enzymatic Activity

Recent changes in fire regimes, with more frequent, extensive, and severe fires, are modifying soil characteristics. The aim of this study was to evaluate the effect of burn severity on the resistance of some physical, chemical, and biochemical soil properties in three different forest ecosystems affected by a wildfire in the northwest of the Iberian Peninsula. We evaluated burn severity immediately after fire using the Composite Burn Index (CBI) in three different ecosystems: shrublands, heathlands, and oak forests. In the same field plots used to quantify CBI, we took a composite soil sample to analyse physical (mean weight diameter (MWD)), chemical (pH; total C; total organic C (TOC); total inorganic C (TIC); total N; available P; exchangeable cations Na+, K+, Mg2+, and Ca2+; and cation exchange capacity (CEC)), and biochemical (β-glucosidase, urease, and acid phosphatase enzyme activities) properties. The resistance index of each property was then calculated. Based on our results, the values of the soil chemical properties tended to increase immediately after fire. Among them, total C, TOC, and exchangeable Na+ showed higher resistance to change, with less variation concerning pre-fire status. The resistance of chemical properties was higher in the oak forest ecosystem. MWD decreased at high severity in all ecosystems, but soils in shrublands were more resistant. We found a high decrease in soil enzymatic activity with burn severity, with biochemical properties being the least resistant to change. Therefore, the enzymatic activity of soil could be a potential indicator of severity in forest ecosystems recently affected by wildfires.

scholarly journals High-Throughput Sequencing Reveals Further Diversity of Little Cherry Virus 1 with Implications for Diagnostics

Viruses ◽  
2018 ◽  
Vol 10 (7) ◽  
pp. 385 ◽  
Author(s):  
Asimina Katsiani ◽  
Varvara Maliogka ◽  
Nikolaos Katis ◽  
Laurence Svanella-Dumas ◽  
Antonio Olmos ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Sour Cherry ◽  
Prunus Species ◽  
Cherry Tree ◽  
High Genetic Diversity ◽  
Viral Genomes ◽  
Virus Diversity ◽  
First Time

Little cherry virus 1 (LChV1, Velarivirus, Closteroviridae) is a widespread pathogen of sweet or sour cherry and other Prunus species, which exhibits high genetic diversity and lacks a putative efficient transmission vector. Thus far, four distinct phylogenetic clusters of LChV1 have been described, including isolates from different Prunus species. The recent application of high throughput sequencing (HTS) technologies in fruit tree virology has facilitated the acquisition of new viral genomes and the study of virus diversity. In the present work, several new LChV1 isolates from different countries were fully sequenced using different HTS approaches. Our results reveal the presence of further genetic diversity within the LChV1 species. Interestingly, mixed infections of the same sweet cherry tree with different LChV1 variants were identified for the first time. Taken together, the high intra-host and intra-species diversities of LChV1 might affect its pathogenicity and have clear implications for its accurate diagnostics.

Extracellular ligation-dependent CD45RB enzymatic activity negatively regulates lipid raft signal transduction

Blood ◽  
2009 ◽  
Vol 113 (3) ◽  
pp. 594-603 ◽  
Author(s):  
Kaushal Parikh ◽  
Sibrand Poppema ◽  
Maikel P. Peppelenbosch ◽  
Lydia Visser
Keyword(s):  
Signal Transduction ◽  
Enzymatic Activity ◽  
Lipid Raft ◽  
Intracellular Signaling ◽  
Tyrosine Phosphatase ◽  
Cell Receptor ◽  
Inhibitory Effect ◽  
Tyrosine Phosphatases ◽  
Peptide Arrays ◽  
Substrate Peptide

Abstract CD45 is the most prominent membrane protein on lymphocytes. The function and regulation of this protein tyrosine phosphatase remain largely obscure, mainly because of the lack of a known ligand, and it still remains unknown whether such tyrosine phosphatases are subject to extracellular control at all. We report that an anti-CD45RB antibody (Ab) that prevents rejection and induces tolerance activates CD45RB tyrosine phosphatase enzymatic activity in T lymphocytes, allowing us to directly monitor the effects of increased CD45RB activity on signal transduction. Using both kinase substrate peptide arrays as well as conventional biochemistry, we also provide evidence of the various kinases involved in bringing about the inhibitory effect of this Ab on CD3-induced T-cell receptor signaling. Furthermore, we report that activated CD45RB translocates to lipid rafts and interferes with lipid raft localization and activation state of CD45 substrate Lck. Thus, these findings indeed prove that CD45 is subject to extracellular control and also define a novel mechanism by which receptor tyrosine phosphatases control lymphocyte biology and provide further insight into the intracellular signaling pathways effected by anti-CD45RB monoclonal Ab treatment.

scholarly journals In Vitro Enzymatic Activity Assays Implicate the Existence of the Chlorophyll Cycle in Chlorophyll b-Containing Cyanobacteria

2019 ◽  
Vol 60 (12) ◽  
pp. 2672-2683 ◽  
Author(s):  
HyunSeok Lim ◽  
Ayumi Tanaka ◽  
Ryouichi Tanaka ◽  
Hisashi Ito
Keyword(s):  
Enzymatic Activity ◽  
Chlorophyll A ◽  
Developmental Stages ◽  
Enzymatic Activities ◽  
Light Acclimation ◽  
Chlorophyll B ◽  
Light Conditions ◽  
Chl A ◽  
Shallow Seas

Abstract In plants, chlorophyll (Chl) a and b are interconvertible by the action of three enzymes—chlorophyllide a oxygenase, Chl b reductase (CBR) and 7-hydroxymethyl chlorophyll a reductase (HCAR). These reactions are collectively referred to as the Chl cycle. In plants, this cyclic pathway ubiquitously exists and plays essential roles in acclimation to different light conditions at various developmental stages. By contrast, only a limited number of cyanobacteria species produce Chl b, and these include Prochlorococcus, Prochloron, Prochlorothrix and Acaryochloris. In this study, we investigated a possible existence of the Chl cycle in Chl b synthesizing cyanobacteria by testing in vitro enzymatic activities of CBR and HCAR homologs from Prochlorothrix hollandica and Acaryochloris RCC1774. All of these proteins show respective CBR and HCAR activity in vitro, indicating that both cyanobacteria possess the potential to complete the Chl cycle. It is also found that CBR and HCAR orthologs are distributed only in the Chl b-containing cyanobacteria that habitat shallow seas or freshwater, where light conditions change dynamically, whereas they are not found in Prochlorococcus species that usually habitat environments with fixed lighting. Taken together, our results implicate a possibility that the Chl cycle functions for light acclimation in Chl b-containing cyanobacteria.

scholarly journals Inactivation of Protein Tyrosine Phosphatases by Peracids Correlates with the Hydrocarbon Chain Length

2015 ◽  
Vol 36 (3) ◽  
pp. 1069-1083 ◽  
Author(s):  
Alicja Kuban-Jankowska ◽  
Magdalena Gorska ◽  
Jack A. Tuszynski ◽  
Cassandra D. M. Churchill ◽  
Philip Winter ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Enzymatic Activity ◽  
Chain Length ◽  
Active Site ◽  
Protein Tyrosine Phosphatases ◽  
Hydrocarbon Chain ◽  
Inhibitory Effect ◽  
Tyrosine Phosphatases ◽  
Medium Chain ◽  
Hydrocarbon Chain Length

Background/Aims: Protein tyrosine phosphatases are crucial enzymes controlling numerous physiological and pathophysiological events and can be regulated by oxidation of the catalytic domain cysteine residue. Peracids are highly oxidizing compounds, and thus may induce inactivation of PTPs. The aim of the present study was to evaluate the inhibitory effect of peracids with different length of hydrocarbon chain on the activity of selected PTPs. Methods: The enzymatic activity of human CD45, PTP1B, LAR, bacterial YopH was assayed under the cell-free conditions, and activity of cellular CD45 in human Jurkat cell lysates. The molecular docking and molecular dynamics were performed to evaluate the peracids binding to the CD45 active site. Results: Here we demonstrate that peracids reduce enzymatic activity of recombinant CD45, PTP1B, LAR, YopH and cellular CD45. Our studies indicate that peracids are more potent inhibitors of CD45 than hydrogen peroxide (with an IC50 value equal to 25 nM for peroctanoic acid and 8 µM for hydrogen peroxide). The experimental data show that the inactivation caused by peracids is dependent on hydrocarbon chain length of peracids with maximum inhibitory effect of medium-chain peracids (C8-C12 acyl chain), which correlates with calculated binding affinities to the CD45 active site. Conclusion: Peracids are potent inhibitors of PTPs with the strongest inhibitory effect observed for medium-chain peracids.

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