scholarly journals Detection and quantification of alkaline phosphatase in single cells of phosphorus-starved marine phytoplankton

1998 ◽  
Vol 164 ◽  
pp. 21-35 ◽  
Author(s):  
S González-Gil ◽  
BA Keafer ◽  
RVM Jovine ◽  
A Aguilera ◽  
S Lu ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Single Cells ◽  
Marine Phytoplankton ◽  
Detection And Quantification

scholarly journals Non-Conventional Metal Ion Cofactor Requirement of Dinoflagellate Alkaline Phosphatase and Translational Regulation by Phosphorus Limitation

2019 ◽  
Vol 7 (8) ◽  
pp. 232 ◽  
Author(s):  
Xin Lin ◽  
Chentao Guo ◽  
Ling Li ◽  
Tangcheng Li ◽  
Senjie Lin
Keyword(s):  
Alkaline Phosphatase ◽  
Metal Ion ◽  
Translational Regulation ◽  
Organic Phosphorus ◽  
Niche Differentiation ◽  
Phosphorus Limitation ◽  
Marine Phytoplankton ◽  
Phosphate Limitation ◽  
Dissolved Organic Phosphorus ◽  
Metal Cofactor

Alkaline phosphatase (AP) enables marine phytoplankton to utilize dissolved organic phosphorus (DOP) when dissolved inorganic phosphate (DIP) is depleted in the ocean. Dinoflagellate AP (Dino-AP) represents a newly classified atypical type of AP, PhoAaty. Despite While being a conventional AP, PhoAEC is known to recruit Zn2+ and Mg2+ in the active center, and the cofactors required by PhoAaty have been contended and remain unclear. In this study, we investigated the metal ion requirement of AP in five dinoflagellate species. After AP activity was eliminated by using EDTA to chelate metal ions, the enzymatic activity could be recovered by the supplementation of Ca2+, Mg2+ and Mn2+ in all cases but not by that of Zn2+. Furthermore, the same analysis conducted on the purified recombinant ACAAP (AP of Amphidinium carterae) verified that the enzyme could be activated by Ca2+, Mg2+, and Mn2+ but not Zn2+. We further developed an antiserum against ACAAP, and a western blot analysis using this antibody showed a remarkable up-regulation of ACAAP under a phosphate limitation, consistent with elevated AP activity. The unconventional metal cofactor requirement of Dino-AP may be an adaptation to trace metal limitations in the ocean, which warrants further research to understand the niche differentiation between dinoflagellates and other phytoplankton that use Zn–Mg AP in utilizing DOP.

scholarly journals On the use of antibiotics for isolating bacteria-free cultures of marine phytoplankton organisms

1952 ◽  
Vol 31 (1) ◽  
pp. 97-106 ◽  
Author(s):  
C. P. Spencer
Keyword(s):  
Fresh Water ◽  
Solid Medium ◽  
Classical Method ◽  
Single Cells ◽  
Marine Phytoplankton ◽  
Marine Diatom ◽  
Sterile Medium ◽  
Kinetics Of ◽  
Water Species ◽  
Algal Cultures

In the course of some studies on the kinetics of growth and the biochemical activities of a marine diatom it became desirable to obtain bacteria-free cultures. The classical method of obtaining such pure algal cultures involves either repeatedly washing single cells in sterile medium or obtaining discrete bacteria-free algal colonies by growth on a solid medium. Both these methods have been widely applied to fresh-water species by Pringsheim (1946) and others, whilst Chu (1946) has used both methods with the marine diatom Nitzschia closterium (Ehrenberg) Wm. Smith forma minutissima.

scholarly journals Localization of human pituitary hormones by multiple immunoenzyme staining procedures using monoclonal and polyclonal antibodies.

1986 ◽  
Vol 34 (3) ◽  
pp. 287-292 ◽  
Author(s):  
S Van Noorden ◽  
M C Stuart ◽  
A Cheung ◽  
E F Adams ◽  
J M Polak
Keyword(s):  
Alkaline Phosphatase ◽  
Binding Sites ◽  
Polyclonal Antibodies ◽  
Single Cells ◽  
Pituitary Hormones ◽  
Tissue Binding ◽  
Human Pituitary ◽  
Single Section ◽  
End Products ◽  
Species Specific

Mouse monoclonal and rabbit polyclonal antibodies to human pituitary hormones were applied together to sections of normal and neoplastic human pituitary tissue. Binding sites were revealed with species-specific immune reagents combined with various enzymes (peroxidase, alkaline phosphatase, and beta-D-galactosidase). The enzymes were developed separately to give differently colored end-products. Where two hormones were present in the same cell, a mixed color was produced. Up to four hormones could be immunostained in a single section. Multiple immunoenzymatic staining has great potential for the analysis of plural antigen production by single cells and relationships between cells producing different antigens.

scholarly journals Device for quantitative cytochemistry: a computerized scanning microdensitometer 'grain counter'.

1978 ◽  
Vol 26 (10) ◽  
pp. 792-802 ◽  
Author(s):  
N S Markovic ◽  
L E Lipkin ◽  
O S Markovic ◽  
M J Wade
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Enzyme Kinetics ◽  
Single Cell ◽  
Quantitative Data ◽  
Single Cells ◽  
Final Reaction Product ◽  
Chamber Studies ◽  
Kinetics Of ◽  
Quantitative Cytochemistry

A computerized scanning microdensitometer and autoradiographic grain counter was able to provide quantitative data on the cytochemical final reaction product formed within a single cell and also quantitate the kinetics of its formation. Optical density and area measurements were performed on hundreds of leukocytes from slides previously stained to demonstrate any one of a variety of reactions. These included cellular glycogen, lipids, peroxidase, esterases, alkaline phosphatase, and acid phosphatase. In addition to these slide studies, chamber studies with an adapted Dvorak-Stotler Chamber allowed the measurement of enzyme kinetics within single cells.

scholarly journals Variable inter and intraspecies alkaline phosphatase activity within single cells of revived dinoflagellates

2021 ◽  
Author(s):  
Mathias Girault ◽  
Raffaele Siano ◽  
Claire Labry ◽  
Marie Latimier ◽  
Cécile Jauzein ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Single Cells

scholarly journals Response of <i>Nodularia spumigena</i> to <i>p</i>CO<sub>2</sub> – Part 2: Exudation and extracellular enzyme activities

2012 ◽  
Vol 9 (4) ◽  
pp. 5109-5151 ◽  
Author(s):  
S. Endres ◽  
J. Unger ◽  
N. Wannicke ◽  
M. Nausch ◽  
M. Voss ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Organic Matter ◽  
Enzyme Activities ◽  
Extracellular Enzymes ◽  
Extracellular Enzyme ◽  
Marine Phytoplankton ◽  
High Pco2 ◽  
Nodularia Spumigena ◽  
Extracellular Enzyme Activities ◽  
Pco2 Treatment

Abstract. The filamentous and diazotrophic cyanobacterium Nodularia spumigena plays a major role in the productivity of the Baltic Sea as it forms extensive blooms regularly. Under phosphorus limiting conditions Nodularia spumigena has a high enzyme affinity for dissolved organic phosphorus (DOP) by production and release of alkaline phosphatase. Additionally, it is able to degrade proteinaceous compounds by expressing the extracellular enzyme leucine aminopeptidase. As atmospheric CO2 concentrations are increasing, we expect marine phytoplankton to experience changes in several environmental parameters including pH, temperature, and nutrient availability. The aim of this study was to investigate the combined effect of CO2-induced changes in seawater carbonate chemistry and of phosphate deficiency on the exudation of organic matter, and its subsequent recycling by extracellular enzymes in a Nodularia spumigena culture. Batch cultures of Nodularia spumigena were grown for 15 days aerated with three different pCO2 levels corresponding to values from glacial periods to future values projected for the year 2100. Extracellular enzyme activities as well as changes in organic and inorganic compound concentrations were monitored. CO2 treatment–related effects were identified for cyanobacterial growth, which in turn was influencing exudation and recycling of organic matter by extracellular enzymes. Biomass production was increased by 56.5% and 90.7% in the medium and high pCO2 treatment, respectively, compared to the low pCO2 treatment and simultaneously increasing exudation. During the growth phase significantly more mucinous substances accumulated in the high pCO2 treatment reaching 363 μg Gum Xanthan eq l−1 compared to 269 μg Gum Xanthan eq l−1 in the low pCO2 treatment. However, cell-specific rates did not change. After phosphate depletion, the acquisition of P from DOP by alkaline phosphatase was significantly enhanced. Alkaline phosphatase activities were increased by factor 1.64 and 2.25, respectively, in the medium and high compared to the low pCO2 treatment. In conclusion, our results suggest that Nodularia spumigena can grow faster under elevated pCO2 by enhancing the recycling of organic matter to acquire nutrients.

Quantitative assays of enzyme activity in single cells: early prenatal diagnosis of genetic disorders.

1977 ◽  
Vol 23 (8) ◽  
pp. 1476-1484 ◽  
Author(s):  
P Hösli
Keyword(s):  
Alkaline Phosphatase ◽  
Gene Dosage ◽  
Clinical Chemistry ◽  
Single Cells ◽  
Genetic Disorders ◽  
Cell Protein ◽  
Lysosomal Storage ◽  
Chromosomal Disorders ◽  
Cell Culture Techniques ◽  
Culture Techniques

Abstract The combined use of special cell culture techniques and biochemical ultramicromethods permits one to handle very small amounts of materials, to reduce the costs of chemicals, and more accurately to assess gene dosage effects by expressing enzyme activities per cell instead of per total cell protein. An alkaline phosphatase induction test has been developed which allows one to screen small numbers of fibroblasts for lysosomal storage diseases, cystic fibrosis, and chromosomal disorders. A successful attempt has been made to automate the microtechniques. Combining the alkaline phosphatase induction with the ultramicro automatization should eventually permit one to screen all pregnancies for major possible fetal genetic defects. Automated ultramicro enzyme assays should contribute to the general development of clinical chemistry.

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