Sensitive Luminometric Method for Protein Quantification in Bacterial Cell Lysate Based on Particle Adsorption and Dissociation of Chelated Europium

2012 ◽  
Vol 84 (3) ◽  
pp. 1386-1393 ◽  
Author(s):  
Sari Pihlasalo ◽  
Antti Kulmala ◽  
Anita Rozwandowicz-Jansen ◽  
Pekka Hänninen ◽  
Harri Härmä
Keyword(s):  
Bacterial Cell ◽  
Protein Quantification ◽  
Cell Lysate ◽  
Bacterial Cell Lysate ◽  
Particle Adsorption ◽  
Adsorption And Dissociation

scholarly journals Diffusion NMR and Rheology Study of a Model Polymer and a Disordered Protein in the Presence of Bacterial Cell Lysate Crowders

2021 ◽  
Vol 120 (3) ◽  
pp. 309a
Author(s):  
Yanitza Trosel ◽  
Valerie Booth ◽  
Anand Yethiraj
Keyword(s):  
Bacterial Cell ◽  
Disordered Protein ◽  
Cell Lysate ◽  
Diffusion Nmr ◽  
Bacterial Cell Lysate

scholarly journals Culturing of ‘Unculturable’ Subsurface Microbes: Natural Organic Carbon Source Fuels the Growth of Diverse and Distinct Bacteria from Groundwater

2020 ◽  
Author(s):  
Xiaoqin Wu ◽  
Sarah Spencer ◽  
Sara Gushgari-Doyle ◽  
Mon Oo Yee ◽  
Jana Voriskova ◽  
...  
Keyword(s):  
Oleic Acid ◽  
Organic Carbon ◽  
Microbial Communities ◽  
Bacterial Cell ◽  
Distinct Species ◽  
Organic C ◽  
Cell Lysate ◽  
Bacterial Cell Lysate ◽  
Diverse Field ◽  
Terrestrial Subsurface

ABSTRACTThe recovery and cultivation of diverse field-related microorganisms from the terrestrial subsurface environment remains a challenge despite recent advances in modern molecular technology. Here we applied natural organic carbon (C), i.e., sediment-derived natural organic matter (NOM) and bacterial cell lysate, to groundwater microbial communities for a 30-day enrichment incubation, followed by conventional direct-plating for isolation. The groundwater was collected from a background well at the Oak Ridge Reservation Field Research Center, Tennessee. As a comparison, we also included enrichments amended with simple organic C sources, including glucose, acetate, benzoate, oleic acid, cellulose, and mixed vitamins. Our results demonstrate that complex natural organic C sources are more effective in enriching diverse bacterial species from groundwater than simple organic C sources. Microcosms amended with simple organic C (glucose, acetate, benzoate, or oleic acid) show significantly lower biodiversity than unamended control and are dominated by only few phyla such as Proteobacteria and Bacteroidetes. In contrast, microcosms amended with complex natural organic C (sediment NOM or bacterial cell lysate) display significantly higher biodiversity, and enrich distinct species from the phyla that are poorly represented in published culture collections (e.g., Verrucomicrobia, Planctomycetes, and Armatimonadetes). Our subsequent isolation efforts from natural organic C-amended enrichments led to 222 purified bacterial isolates representing 5 phyla, 16 orders, and 54 distinct species including candidate novel, rarely cultivated, and undescribed organisms.ImportanceInnovative strategies for recovering bacterial strains representing the true diversity of microbial communities in the terrestrial subsurface would significantly advance the understanding of ecologically critical taxa residing in these ecosystems. In this study, we demonstrate that complex natural organic C that mimic the naturally available resources for microbes encourages the growth of diverse bacteria much more robustly than traditional simplistic organic C sources. Results from this study will substantially advance and improve the design of strategies to effectively cultivate and isolate diverse and novel subsurface microorganisms in the laboratory. Obtaining axenic cultures of the ‘once-unculturable’ microorganisms will greatly enhance our understanding of microbial physiology, function, and roles in different biogeochemical niches in terrestrial subsurface ecosystems.

scholarly journals Culturing of “Unculturable” Subsurface Microbes: Natural Organic Carbon Source Fuels the Growth of Diverse and Distinct Bacteria From Groundwater

2020 ◽  
Vol 11 ◽  
Author(s):  
Xiaoqin Wu ◽  
Sarah Spencer ◽  
Sara Gushgari-Doyle ◽  
Mon Oo Yee ◽  
Jana Voriskova ◽  
...  
Keyword(s):  
Distinct Species ◽  
Bacterial Isolates ◽  
Culture Collections ◽  
Microbial Physiology ◽  
Cell Lysate ◽  
Bacterial Cell Lysate ◽  
Organic Carbon Source ◽  
Complex Carbon ◽  
And Function ◽  
Terrestrial Subsurface

Recovery and cultivation of diverse environmentally-relevant microorganisms from the terrestrial subsurface remain a challenge despite recent advances in modern molecular technology. Here, we applied complex carbon (C) sources, i.e., sediment dissolved organic matter (DOM) and bacterial cell lysate, to enrich groundwater microbial communities for 30 days. As comparisons, we also included enrichments amended with simple C sources including glucose, acetate, benzoate, oleic acid, cellulose, and mixed vitamins. Our results demonstrate that complex C is far more effective in enriching diverse and distinct microorganisms from groundwater than simple C. Simple C enrichments yield significantly lower biodiversity, and are dominated by few phyla (e.g., Proteobacteria and Bacteroidetes), while microcosms enriched with complex C demonstrate significantly higher biodiversity including phyla that are poorly represented in published culture collections (e.g., Verrucomicrobia, Planctomycetes, and Armatimonadetes). Subsequent isolation from complex C enrichments yielded 228 bacterial isolates representing five phyla, 17 orders, and 56 distinct species, including candidate novel, rarely cultivated, and undescribed organisms. Results from this study will substantially advance cultivation and isolation strategies for recovering diverse and novel subsurface microorganisms. Obtaining axenic representatives of “once-unculturable” microorganisms will enhance our understanding of microbial physiology and function in different biogeochemical niches of terrestrial subsurface ecosystems.

Contribution of large bacteria to bacterial biomass in a deep freshwater lake (Lake Biwa, Japan)

2020 ◽  
Vol 85 ◽  
pp. 131-139
Author(s):  
S Shen ◽  
Y Shimizu
Keyword(s):  
Cell Volume ◽  
Bacterial Cell ◽  
Lake Biwa ◽  
Bacterial Biomass ◽  
Spatial Variations ◽  
Freshwater Lake ◽  
Heterotrophic Nanoflagellates ◽  
Bacterial Productivity ◽  
Transmission Electron ◽  
Seasonal And Spatial Variations

Despite the importance of bacterial cell volume in microbial ecology in aquatic environments, literature regarding the effects of seasonal and spatial variations on bacterial cell volume remains scarce. We used transmission electron microscopy to examine seasonal and spatial variations in bacterial cell size for 18 mo in 2 layers (epilimnion 0.5 m and hypolimnion 60 m) of Lake Biwa, Japan, a large and deep freshwater lake. During the stratified period, we found that the bacterial cell volume in the hypolimnion ranged from 0.017 to 0.12 µm3 (median), whereas that in the epilimnion was less variable (0.016 to 0.033 µm3, median) and much lower than that in the hypolimnion. Additionally, in the hypolimnion, cell volume during the stratified period was greater than that during the mixing period (up to 5.7-fold). These differences in cell volume resulted in comparable bacterial biomass in the hypolimnion and epilimnion, despite the fact that there was lower bacterial abundance in the hypolimnion than in the epilimnion. We also found that the biomass of larger bacteria, which are not likely to be grazed by heterotrophic nanoflagellates, increased in the hypolimnion during the stratified period. Our data suggest that estimation of carbon flux (e.g. bacterial productivity) needs to be interpreted cautiously when cell volume is used as a constant parametric value. In deep freshwater lakes, a difference in cell volume with seasonal and spatial variation may largely affect estimations.

High-throughput Detection of Protein Kinase Activities in Cell Lysate Based on the Aggregation of Gold Nanoparticles with Peptides

2009 ◽  
Author(s):  
Yoshiki Katayama ◽  
Hirotaro Kitazaki ◽  
Jeong-Hun Kang ◽  
Xiaoming Han ◽  
Takeshi Mori ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Protein Kinase ◽  
High Throughput ◽  
Cell Lysate ◽  
High Throughput Detection

A Photoactivable Formaldehyde Donor with Fluorescence Monitoring Reveals Threshold to Arrest Cell Migration

2019 ◽  
Author(s):  
Lukas P Smaga ◽  
Nicholas W Pino ◽  
Gabriela E Ibarra ◽  
Vishnu Krishnamurthy ◽  
Jefferson Chan
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Fluorescence Enhancement ◽  
Biological Effects ◽  
Cellular Systems ◽  
Live Cells ◽  
First Report ◽  
Cell Lysate ◽  
Intracellular Release ◽  
Biological Analytes

Controlled light-mediated delivery of biological analytes enables the investigation of highly reactivity molecules within cellular systems. As many biological effects are concentration dependent, it is critical to determine the location, time, and quantity of analyte donation. In this work, we have developed the first photoactivatable donor for formaldehyde (FA). Our optimized photoactivatable donor, photoFAD-3, is equipped with a fluorescence readout that enables monitoring of FA release with a concomitant 139-fold fluorescence enhancement. Tuning of photostability and cellular retention enabled quantification of intracellular FA release through cell lysate calibration. Application of photoFAD-3 uncovered the concentration range necessary for arresting wound healing in live cells. This marks the first report where a photoactivatable donor for any analyte has been used to quantify intracellular release.

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