Formulation of Culture Medium for Superoxide Dismutase Production by Rhodotorula Glutinis RY-06 Strain with High Yield of Beta-Carotene

2008 ◽  
Author(s):  
Sui-Lou Wang ◽  
Gui-Tang Chen ◽  
Guo-Hong Qi ◽  
Zhi-Ping Yang ◽  
Bo Li
Keyword(s):  
Superoxide Dismutase ◽  
Culture Medium ◽  
Rhodotorula Glutinis ◽  
Beta Carotene ◽  
High Yield

scholarly journals Defined MSC exosome with high yield and purity to improve regenerative activity

2021 ◽  
Vol 12 ◽  
pp. 204173142110086
Author(s):  
Jun Yong Kim ◽  
Won-Kyu Rhim ◽  
Yong-In Yoo ◽  
Da-Seul Kim ◽  
Kyoung-Won Ko ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Density Lipoprotein ◽  
High Yield ◽  
Human Umbilical Cord ◽  
Cell Culture Medium ◽  
Human Coronary Artery ◽  
Isolation Methods ◽  
Regenerative Activity ◽  
Yield And Purity

Exosomes derived from mesenchymal stem cells (MSCs) have been studied as vital components of regenerative medicine. Typically, various isolation methods of exosomes from cell culture medium have been developed to increase the isolation yield of exosomes. Moreover, the exosome-depletion process of serum has been considered to result in clinically active and highly purified exosomes from the cell culture medium. Our aim was to compare isolation methods, ultracentrifuge (UC)-based conventional method, and tangential flow filtration (TFF) system-based method for separation with high yield, and the bioactivity of the exosome according to the purity of MSC-derived exosome was determined by the ratio of Fetal bovine serum (FBS)-derived exosome to MSC-derived exosome depending on exosome depletion processes of FBS. The TFF-based isolation yield of exosome derived from human umbilical cord MSC (UCMSC) increased two orders (92.5 times) compared to UC-based isolation method. Moreover, by optimizing the process of depleting FBS-derived exosome, the purity of UCMSC-derived exosome, evaluated using the expression level of MSC exosome surface marker (CD73), was about 15.6 times enhanced and the concentration of low-density lipoprotein-cholesterol (LDL-c), known as impurities resulting from FBS, proved to be negligibly detected. The wound healing and angiogenic effects of highly purified UCMSC-derived exosomes were improved about 23.1% and 71.4%, respectively, with human coronary artery endothelial cells (HCAEC). It suggests that the defined MSC exosome with high yield and purity could increase regenerative activity.

Effect of cerium dioxide nanoparticles on certain indication of synthetic activity Rhodotorula glutinis

2020 ◽  
Vol 12 (2) ◽  
pp. 166-173
Author(s):  
Anastasiia Obradovych ◽  
Liliia Vasina ◽  
Nadiia Zholobak
Keyword(s):  
Cerium Dioxide ◽  
Culture Medium ◽  
Antioxidant Properties ◽  
Rhodotorula Glutinis ◽  
Component Composition ◽  
Biologically Active Compounds ◽  
Biologically Active ◽  
Synthetic Activity ◽  
Yeast Saccharomyces Cerevisiae ◽  
Active Compounds

Cerium dioxide nanoparticles (CeO2 NP) have significant catalytic and antioxidant properties due to their ability to inhibit the development of free-radical reactions due to a combination of superoxidedismutase and catalase mimetic activities. This nanopreparation has a significant antibacterial effect, but is low-toxic to unicellular yeast Saccharomyces cerevisiae. Therefore, it is relevant and appropriate to study the effect of this compound on other groups of yeast, in particular members of the genus Rhodotorula spp., which are characterized by high growth rate, unpretentiousness to the component composition of the culture medium, the ability to form a variety of valuable biologically active compounds, as well as the ability to synthesize proteins, lipids, carotenoids and exopolysaccharides in the complex. The report that microorganisms of this genus have been identified as part of the normal fish microbiota (Abramis brama, Rutilus rutilus, Perca fluviatilis, Oreochromis niloticus) expands the range of their use, either as probiotics in general or their individual cellular and extracellular components as prebiotics. In this work, the effect of different concentrations of CeO2 NP nanopreparation on the production parameters of Rhodotorula glutinis was studied, as well as their antagonistic properties after saturation of nanocerium were tested. To detect the optimal concentration of nanopreparation of cerium dioxide, R. glutinis was cultured for 120 h at 28 0C and aerated at 160 rpm on Saburo medium with the addition of citrate-stabilized CeO2 NP with a particle size of 1-2 nm in the following concentrations: 100 mM, 10 mM, 1 mM, 0,1 mM, 0,01 mM. The control was R. glutinis grown on Saburo medium, experimental controls were represented by microorganisms in the culture medium of which citrate and full-size cerium (CeCl3) were added. It was found that most of the studied concentrations of CeO2 NP (except for the highest) are not toxic to these carotenogenic yeasts. The presence of 10 mM CeO2 NP stimulates the growth of the culture, while the increase in biomass is increased by fourfold compared to the control. There was also an increase in the content of basic metabolites – protein by 1,2 times, lipids by 2,5 times and valuable biologically active compounds - carotenoids by 20% and exopolysaccharides by 2,2 times. However, under these conditions, no significant changes in catalase and superoxidase activities were registered. Examination of the antagonistic properties of the CeO2 NP-enriched culture of R. glutinis by delayed antagonism by the method of perpendicular strokes against gram-positive B. subtilis and gram-negative P. syringae bacteria showed the presence of lysis zones up to 17 and 25 mm, respectively, in contrast to their complete or complete absence (2 mm), noted in the study of control samples. A study of the resistance of CeO2 NP-enriched culture of R. glutinis to natural inhibitors of the digestive tract showed resistance of microorganisms to gastric juice (the amount of CFU is 1,4 times higher than control values) and bile (recorded a 5-fold increase in survival of microorganisms). The obtained results testify to the prospects of using nanocerium for mobilization of growth, enzymatic, antagonistic activities of R. glutinis culture.

High yield of human umbilical cord-derived mesenchymal stromal cells in an animal component-free culture medium

Cytotherapy ◽  
2020 ◽  
Vol 22 (5) ◽  
pp. S111-S112
Author(s):  
R. Wagey ◽  
K. Bertram ◽  
M. Elliott ◽  
A. Eaves ◽  
S. Szilvassy ◽  
...  
Keyword(s):  
Umbilical Cord ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Culture Medium ◽  
High Yield ◽  
Human Umbilical Cord ◽  
Animal Component

scholarly journals Human extracellular superoxide dismutase is a tetramer composed of two disulphide-linked dimers: a simplified, high-yield purification of extracellular superoxide dismutase

1996 ◽  
Vol 317 (1) ◽  
pp. 51-57 ◽  
Author(s):  
Tim D. OURY ◽  
James D. CRAPO ◽  
Zuzana VALNICKOVA ◽  
Jan J. ENGHILD
Keyword(s):  
Superoxide Dismutase ◽  
Limited Proteolysis ◽  
Exchange Chromatography ◽  
Disulphide Bond ◽  
High Yield ◽  
Human Aorta ◽  
Extracellular Superoxide Dismutase ◽  
Heparin Binding ◽  
Sod Activity ◽  
Native Page

Studies examining the biochemical characteristics and pharmacological properties of extracellular superoxide dismutase (EC SOD) have been severely limited because of difficulties in purifying the enzyme. Recently EC SOD was found to exist in high concentrations in the arteries of most mammals examined and it is the predominant form of SOD activity in many arteries. We now describe a three-step, high-yield protocol for the purification of EC SOD from human aorta. In the first step, the high affinity of EC SOD for heparin is utilized to obtain a fraction in which EC SOD constitutes roughly 13% of the total protein compared with only 0.3% of that of the starting material. In addition, over 80% of the original EC SOD activity present in the aortic homogenate was retained after the first step of purification. EC SOD was further purified using a combination of cation- and anion-exchange chromatography. The overall yield of EC SOD from this purification procedure was 46%, with over 4 mg of EC SOD obtained from 230 g of aorta. Purified EC SOD was found to exist predominantly as a homotetramer composed of two disulphide-linked dimers. However, EC SOD was also found to form larger multimers when analysed by native PAGE. It was shown by urea denaturation that the formation of multimers increased the thermodynamic stability of the protein. Limited proteolysis of EC SOD suggested that there is one interchain disulphide bond covalently linking two subunits. This disulphide bond involves cysteine-219 and appears to link the heparin-binding domains of the two subunits.

Kinetics of a rapid, luminol dependent chemiluminescence signal induced in HL-60 cells by amphotericin B and other stimulants

1991 ◽  
Vol 69 (9) ◽  
pp. 618-623 ◽  
Author(s):  
D. A. O'Keefe ◽  
D. R. James ◽  
W. R. Ware ◽  
N. O. Petersen
Keyword(s):  
Tissue Culture ◽  
Superoxide Dismutase ◽  
Amphotericin B ◽  
Culture Medium ◽  
Polyene Antibiotic ◽  
Biochemical Processes ◽  
Rate Determining Step ◽  
Rate Limiting ◽  
Kinetics Of ◽  
Peak Value

Addition of the polyene antibiotic amphotericin B or tissue culture medium to nondifferentiated HL-60 cells in the presence of luminol induces a chemiluminescence signal that reaches a peak value within a few seconds and decays exponentially in less than a minute. The kinetics of the signal and its modulation by superoxide dismutase, catalase, and horseradish peroxidase are consistent with a series of solution biochemical processes with a rate-determining step corresponding to the disproportionation of a luminal–superoxide complex. The effects of the enzymes demonstrate that superoxide is a precursor to the rate-determining intermediate and that both catalase and peroxide enhance a reaction that competes with the rate-limiting process.Key words: chemiluminescence, luminol, amphotericin B, superoxide, HL-60 cells.

A Method of Obtaining a Suspension of Intact Parenchymal Cells from Adult Rat Liver

1971 ◽  
Vol 8 (1) ◽  
pp. 73-86
Author(s):  
JENNIFER J. GALLAI-HATCHARD ◽  
G. M. GRAY
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Culture Medium ◽  
High Yield ◽  
Alkaline Ph ◽  
High Osmolarity ◽  
Adult Rat ◽  
Parenchymal Cells ◽  
Phosphate Buffered Saline

The perfusion of liver with either citrate or tetraphenyl boron to remove Ca2+ or K+ or with a solution of high osmolarity and alkaline pH yields plenty of cells but they are all damaged. Perfusion of the liver with hyaluronidase and collagenase followed by incubation of liver slices in the same enzyme solution produced a high yield of cells (25%, w/w, of liver) of which only about 1% were undamaged. However, perfusion with 0.3% hyaluronidase, 0.3% collagenase and 0.1% trypsin in phosphate-buffered saline (excluding Mg2+ and Ca2+) followed by incubation at 25 °C of the chopped liver gave a small yield (2-4%, w/w) of undamaged cells which were not permeable to eosin for up to an hour when suspended in culture medium containing 2% bovine serum albumin.

Screening of Lipid High Producing Mutant from Rhodotorula Glutinis by Low Ion Implantation and Study on Lipid Extration Technique

2011 ◽  
Vol 343-344 ◽  
pp. 1172-1181
Author(s):  
Shi Chang Li ◽  
Hong Xia Liu ◽  
Shao Bin Gu ◽  
Min Wang ◽  
Zhao Yang Zhu ◽  
...  
Keyword(s):  
Ion Implantation ◽  
Treatment Time ◽  
Lipid Extraction ◽  
Rhodotorula Glutinis ◽  
High Yield ◽  
Fermentation Time ◽  
Lipid Yield ◽  
Ultrasonic Time ◽  
Burman Design ◽  
Plackett Burman Design

The paper studied on parameters of ion implantation into lipid producing strain Rhodotorula glutinis and lipid extration technology. It was found that the strain had a higher positive mutation rate when the output power was 10keV and the dose of N+ implantation was 80×2.6×1013 ions/cm2. Then a high-yield mutant strain D30 was obtained and it’s lipid yield which was 3.10 g/L increased by 33.05% than that of the original. Additionally, statistically-based experimental designs were applied for the optimization of lipid extraction by acid-heating coupling ultrasonic technique. By a Plackett-Burman design, it was found that three factors, treatment time of HCl (p=0.036) , ultrasonic time (p=0.0105) and rate of extracting solution (VCHCl3:VCH3OH) (p=0.0105), had significant effect on lipid extraction. Subsequently, these significant factors were optimized using response surface methodology (RSM), and the optimized parameters of lipid extraction were as follows: 34 min for treatment time of HCl, 7.5 min for ultrasonic treating time, 1.9:1 for rate of extracting solution (VCHCl3:VCH3OH). Finally the fermentation characteristic of high-yield mutation strain D30 was studied, when fermentation time was 10 d, it’s lipid yield increased to 7.81 g/L

scholarly journals Superoxide-dependent consumption of nitric oxide in biological media may confound in vitro experiments

2003 ◽  
Vol 369 (2) ◽  
pp. 399-406 ◽  
Author(s):  
Robert G. KEYNES ◽  
Charmaine GRIFFITHS ◽  
John GARTHWAITE
Keyword(s):  
Tissue Culture ◽  
Superoxide Dismutase ◽  
Culture Medium ◽  
Superoxide Radical ◽  
Cultured Cells ◽  
Soluble Guanylate Cyclase ◽  
Subsequent Reaction ◽  
Biological Media ◽  
State Concentration

NO functions ubiquitously as a biological messenger but has also been implicated in various pathologies, a role supported by many reports that exogenous or endogenous NO can kill cells in tissue culture. In the course of experiments aimed at examining the toxicity of exogenous NO towards cultured cells, we found that most of the NO delivered using a NONOate (diazeniumdiolate) donor was removed by reaction with the tissue-culture medium. Two NO-consuming ingredients were identified: Hepes buffer and, under laboratory lighting, the vitamin riboflavin. In each case, the loss of NO was reversed by the addition of superoxide dismutase. The effect of Hepes was observed over a range of NONOate concentrations (producing up to 1μM NO). Furthermore, from measurements of soluble guanylate cyclase activity, Hepes-dependent NO consumption remained significant at the low nanomolar NO concentrations relevant to physiological NO signalling. The combination of Hepes and riboflavin (in the light) acted synergistically to the extent that, instead of a steady-state concentration of about 1μM being generated, NO was undetectable (<10nM). Again, the consumption could be inhibited by superoxide dismutase. A scheme is proposed whereby a ‘vicious cycle’ of superoxide radical (O2•-) formation occurs as a result of oxidation of Hepes to its radical species, fuelled by the subsequent reaction of O2•- with NO to form peroxynitrite (ONOO-). The inadvertent production of ONOO- and other reactive species in biological media, or the associated loss of NO, may contribute to the adverse effects, or otherwise, of NO in vitro.

scholarly journals Cytosolic iron superoxide dismutase is a part of the triacylglycerol biosynthetic complex in oleaginous yeast

2003 ◽  
Vol 372 (2) ◽  
pp. 587-594 ◽  
Author(s):  
Sumana RAYCHAUDHURI ◽  
Mamatha M. REDDY ◽  
Naveen R. RAJKUMAR ◽  
Ram RAJASEKHARAN
Keyword(s):  
Superoxide Dismutase ◽  
Oleaginous Yeast ◽  
Rhodotorula Glutinis ◽  
Soluble Form ◽  
Nucleotide Sequence Analysis ◽  
Iron Superoxide Dismutase ◽  
Oxidative Damages ◽  
Cdna Nucleotide Sequence ◽  
Protein Components ◽  
First Time

A novel multienzyme complex for the biosynthesis of triacylglycerol in oleaginous yeast has been identified recently in the cytosol and characterized [Gangar, Karande and Rajasekharan (2001) J. Biol. Chem. 276, 10290–10298]. Screening the library of Rhodotorula glutinis with an oligonucleotide probe derived from the N-terminal sequence of one of the protein components in the complex (21 kDa protein) resulted in the isolation of a 0.7 kb cDNA. Nucleotide sequence analysis revealed that the isolated gene codes for superoxide dismutase (SOD). Atomic absorption spectroscopy and inhibition assays showed that this cytosolic SOD utilizes Fe as its cofactor. Enzymic assays, immunoprecipitation and cross-linking experiments revealed that SOD is a part of the triacylglycerol biosynthetic complex, which could protect the substrate and the complex from oxidative damages. These results indicate for the first time the presence of iron-containing SOD in a soluble form in yeast.

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