Enzyme Fractionation by Means of Large- Scale Continuous- Flow Electrophoresis

1966 ◽  
Vol 12 (8) ◽  
pp. 497-504 ◽  
Author(s):  
Seymour Winsten ◽  
Jeanne Jackson ◽  
Paula Wolf
Keyword(s):  
Lactic Acid ◽  
Enzyme Activities ◽  
Continuous Flow ◽  
Large Scale ◽  
Leucine Aminopeptidase ◽  
Ph Effect ◽  
Good Recovery ◽  
Acid Dehydrogenase ◽  
Aldolase Activity ◽  
Single Serum Sample

Abstract Large- scale, microglass bead, continuous- flow electrophoresis was used to separateand collect protein fractions from a single serum sample. These fractions were assayed for a multiplicity of enzyme activities. Relatively good recovery was obtained with lactic acid dehydrogenase, leucine aminopeptidase, and phosphohexoseisomerase. Artifactually high recoveries were observed with aldotase and isocitric acid dehydrogenase. The increase observed in aldolase activity may be due to a pH effect.

scholarly journals Thermal reaction norms of key metabolic enzymes reflect divergent physiological and behavioral adaptations of closely related amphipod species

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Lena Jakob ◽  
Kseniya P. Vereshchagina ◽  
Anette Tillmann ◽  
Lorena Rivarola-Duarte ◽  
Denis V. Axenov-Gribanov ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Lake Baikal ◽  
Enzyme Activities ◽  
Large Scale ◽  
Metabolic Enzymes ◽  
Thermal Reaction ◽  
Reaction Norms ◽  
Ecological Niches ◽  
Amphipod Species ◽  
Thermal Plasticity

AbstractLake Baikal is inhabited by more than 300 endemic amphipod species, which are narrowly adapted to certain thermal niches due to the high interspecific competition. In contrast, the surrounding freshwater fauna is commonly represented by species with large-scale distribution and high phenotypic thermal plasticity. Here, we investigated the thermal plasticity of the energy metabolism in two closely-related endemic amphipod species from Lake Baikal (Eulimnogammarus verrucosus; stenothermal and Eulimnogammarus cyaneus; eurythermal) and the ubiquitous Holarctic amphipod Gammarus lacustris (eurythermal) by exposure to a summer warming scenario (6–23.6 °C; 0.8 °C d−1). In concert with routine metabolic rates, activities of key metabolic enzymes increased strongly with temperature up to 15 °C in E. verrucosus, whereupon they leveled off (except for lactate dehydrogenase). In contrast, exponential increases were seen in E. cyaneus and G. lacustris throughout the thermal trial (Q10-values: 1.6–3.7). Cytochrome-c-oxidase, lactate dehydrogenase, and 3-hydroxyacyl-CoA dehydrogenase activities were found to be higher in G. lacustris than in E. cyaneus, especially at the highest experimental temperature (23.6 °C). Decreasing gene expression levels revealed some thermal compensation in E. cyaneus but not in G. lacustris. In all species, shifts in enzyme activities favored glycolytic energy generation in the warmth. The congruent temperature-dependencies of enzyme activities and routine metabolism in E. verrucosus indicate a strong feedback-regulation of enzymatic activities by whole organism responses. The species-specific thermal reaction norms reflect the different ecological niches, including the spatial distribution, distinct thermal behavior such as temperature-dependent migration, movement activity, and mating season.

Using Indicator Bacteria and Salmonella Test Results from Three Large-Scale Beef Abattoirs over an 18-Month Period To Evaluate Intervention System Efficacy and Plan Carcass Testing for Salmonella

2007 ◽  
Vol 70 (12) ◽  
pp. 2732-2740 ◽  
Author(s):  
JOHN R. RUBY ◽  
JUN ZHU ◽  
STEVEN C. INGHAM
Keyword(s):  
Lactic Acid ◽  
Large Scale ◽  
Microbial Contamination ◽  
Test Results ◽  
Microbiological Contamination ◽  
Negative Results ◽  
Intervention Process ◽  
Beef Carcasses ◽  
Causative Factors ◽  
Plate Counts

To develop a process for predicting the likelihood of Salmonella contamination on beef carcasses, we evaluated the influence of several possible causative factors (i.e., year, abattoir, day of week, month, and intervention system components) on the risk of Salmonella and indicator organism contamination. Hide and carcass sponge samples were collected in 2005 to 2006 in six steps at three abattoirs in the East (A), Midwest (B), and Southwest (C) United States. Each abattoir used the same intervention system. Samples were analyzed for aerobic plate counts (APCs; n = 18,990) and Enterobacteriaceae counts (EBCs; n = 18,989) and the presence or absence of Salmonella (n = 5,355). Our results demonstrated that many factors play a significant role in the level of microbial contamination of beef carcasses. Overall, Salmonella prevalence and EBC levels were significantly higher in 2006 than in 2005. APCs and EBCs were highest in abattoirs A (3.57 log CFU/100 cm2) and B (1.31 log CFU/100 cm2). The odds of detecting a positive Salmonella isolate were greatest in abattoir C and lowest in abattoir A. Across the three abattoirs, the overall intervention process effectively reduced microbiological contamination. Salmonella prevalence fell from 45% (preevisceration) to 0.47% (postchilled–lactic acid), and there were APC and EBC reductions of 5.43 and 5.28 log CFU/100 cm2, respectively, from hide-on to postchilled–lactic acid samples. At each abattoir, composites of three individual EBC-negative carcass samples yielded Salmonella-negative results 97 to 99% of the time. These results suggest the possibility of using indicator test results to accurately predict the absence of Salmonella in a beef carcass sample.

Optimized Large Scale Synthesis of Phosphorothioate Oligonucleotides on PS-PEG Supports

1997 ◽  
Vol 52 (1) ◽  
pp. 110-116
Author(s):  
Michael Gerster ◽  
Martin Maier ◽  
Nils Clausen ◽  
Jens Schewitz ◽  
Ernst Bayer
Keyword(s):  
Gel Electrophoresis ◽  
Continuous Flow ◽  
Large Scale ◽  
Electrospray Mass Spectrometry ◽  
Small Scale ◽  
Batch Mode ◽  
Crucial Step ◽  
Capillary Gel Electrophoresis ◽  
Phosphoramidite Chemistry ◽  
Large Scale Synthesis

Sulphurization is a crucial step during synthesis of phosphorothioate oligonucleotides. Insufficient reaction leads to inhomogeneous products with phosphodiester defects and subsequently to destabilization of the oligomers in biological media. To achieve a maximum extent of sulphur incorporation, various sulphurizing agents have been investigated. Solely, the use of Beaucage reagent provided satisfactory results on PS-PEG supports. Based on our investigations in small scale synthesis (1 μmol) with continuous-flow technique, upscaling to the 0.1-0.25 mmolar range has been achieved using a peptide synthesizer. The syntheses were performed in batch mode with standard phosphoramidite chemistry. Additionally, large scale synthesis of a phosphodiester oligonucleotide has been carried out on PS-PEG with optimized protocols and compared to small scale synthesis on different supports. Products were analysed by 31P NMR, capillary gel electrophoresis and electrospray mass spectrometry. An extent of sulphurization of 99% and coupling effiencies of more than 99% were obtained and the products proved to have similar purity compared to small scale syntheses on CPG

scholarly journals Review on the Biological Detoxification of Mycotoxins Using Lactic Acid Bacteria to Enhance the Sustainability of Foods Supply

Molecules ◽  
2020 ◽  
Vol 25 (11) ◽  
pp. 2655 ◽  
Author(s):  
Belal J. Muhialdin ◽  
Nazamid Saari ◽  
Anis Shobirin Meor Hussin
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Large Scale ◽  
Negative Impact ◽  
Cost Effective ◽  
Economic Loss ◽  
Current Review ◽  
Good Potential ◽  
Biological Detoxification ◽  
Safe Food

The challenges to fulfill the demand for a safe food supply are dramatically increasing. Mycotoxins produced by certain fungi cause great economic loss and negative impact on the sustainability of food supplies. Moreover, the occurrence of mycotoxins at high levels in foods poses a high health threat for the consumers. Biological detoxification has exhibited a high potential to detoxify foodstuffs on a cost-effective and large scale. Lactic acid bacteria showed a good potential as an alternative strategy for the elimination of mycotoxins. The current review describes the health and economic impacts associated with mycotoxin contamination in foodstuffs. Moreover, this review highlights the biological detoxification of common food mycotoxins by lactic acid bacteria.

scholarly journals Biotechnological valorization of agro industrial and household wastes for lactic acid production

2019 ◽  
Vol 21 (1) ◽  
pp. 113-127 ◽  
Author(s):  
Juliana Romo-Buchelly ◽  
María Rodríguez-Torres ◽  
Fernando Orozco-Sánchez
Keyword(s):  
Lactic Acid ◽  
Large Scale ◽  
Lactic Acid Production ◽  
Scale Production ◽  
Advantages And Disadvantages ◽  
Large Scale Production ◽  
Different Types ◽  
High Transformation ◽  
Transformation Potential ◽  
The Cost

Lactic acid (LA) is an organic compound used in several industries, such as food, textile, chemical, and pharmaceutical. The global interest  in  this  product  is  due  to  its  use  for  the  synthesis  of  numerous  chemical  compounds,  including  polylactic  acid,  a  biode-gradable thermoplastic and substitute for petroleum-derived plastics. An in-depth overview of the use of industrial and household wastes as inexpensive substrates in order to reduce the cost of LA production is presented. A review is carried out of the biotech-nological aspects that must be taken into account when using some wastes with high transformation potential to produce LA in a submerged  culture,  as  well  recommendations  for  their  use.  The  advantages  and  disadvantages  of  different  types  of  treatments used for the transformation of waste into suitable substrates are considered. Several methods of fermentation, as well as genetic strategies for increasing the production, are summarized and compared. It is expected that in a few years there will be many ad-vances in these areas that will allow greater large-scale production of LA using agroindustrial or household wastes, with potential positive economic and environmental impact in some regions of the planet.

scholarly journals Extracellular Enzyme Activity and Microbial Diversity Measured on Seafloor Exposed Basalts from Loihi Seamount Indicate the Importance of Basalts to Global Biogeochemical Cycling

2014 ◽  
Vol 80 (16) ◽  
pp. 4854-4864 ◽  
Author(s):  
Myrna E. Jacobson Meyers ◽  
Jason B. Sylvan ◽  
Katrina J. Edwards
Keyword(s):  
Enzyme Activity ◽  
Microbial Communities ◽  
Enzyme Activities ◽  
Leucine Aminopeptidase ◽  
Extracellular Enzyme ◽  
Basaltic Rock ◽  
Rrna Gene ◽  
Content Type ◽  
Shelf Sediments ◽  
Potential Enzyme Activity

ABSTRACTSeafloor basalts are widely distributed and host diverse prokaryotic communities, but no data exist concerning the metabolic rates of the resident microbial communities. We present here potential extracellular enzyme activities of leucine aminopeptidase (LAP) and alkaline phosphatase (AP) measured on basalt samples from different locations on Loihi Seamount, HI, coupled with analysis of prokaryotic biomass and pyrosequencing of the bacterial 16S rRNA gene. The community maximum potential enzyme activity (Vmax) of LAP ranged from 0.47 to 0.90 nmol (g rock)−1h−1; theVmaxfor AP was 28 to 60 nmol (g rock)−1h−1. TheKmof LAP ranged from 26 to 33 μM, while theKmfor AP was 2 to 7 μM. Bacterial communities on Loihi basalts were comprised primarily ofAlpha-,Delta-, andGammaproteobacteria,Bacteroidetes, andPlanctomycetes. The putative ability to produce LAP is evenly distributed across the most commonly detected bacterial orders, but the ability to produce AP is likely dominated by bacteria in the ordersXanthomonadales,Flavobacteriales, andPlanctomycetales. The enzyme activities on Loihi basalts were compared to those of other marine environments that have been studied and were found to be similar in magnitude to those from continental shelf sediments and orders of magnitude higher than any measured in the water column, demonstrating that the potential for exposed basalts to transform organic matter is substantial. We propose that microbial communities on basaltic rock play a significant, quantifiable role in benthic biogeochemical processes.

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