scholarly journals A Novel Agarase, Gaa16B, Isolated from the Marine Bacterium Gilvimarinus agarilyticus JEA5, and the Moisturizing Effect of Its Partial Hydrolysis Products

Marine Drugs ◽  
2021 ◽  
Vol 20 (1) ◽  
pp. 2
Author(s):  
Youngdeuk Lee ◽  
Eunyoung Jo ◽  
Yeon-Ju Lee ◽  
Tae-Yang Eom ◽  
Yehui Gang ◽  
...  
Keyword(s):  
Marine Bacterium ◽  
Average Molecular Weight ◽  
Biochemical Properties ◽  
Partial Hydrolysis ◽  
Carbohydrate Binding ◽  
Thin Layer Chromatography Analysis ◽  
Amino Acid Similarity ◽  
Chromatography Analysis ◽  
Hydrolysis Products ◽  
Layer Chromatography

We recently identified a β-agarase, Gaa16B, in the marine bacterium Gilvimarinus agarilyticus JEA5. Gaa16B, belonging to the glycoside hydrolase 16 family of β-agarases, shows less than 70.9% amino acid similarity with previously characterized agarases. Recombinant Gaa16B lacking the carbohydrate-binding region (rGaa16Bc) was overexpressed in Escherichia coli and purified. Activity assays revealed the optimal temperature and pH of rGaa16Bc to be 55 ∘C and pH 6–7, respectively, and the protein was highly stable at 55 ∘C for 90 min. Additionally, rGaa16Bc activity was strongly enhanced (2.3-fold) in the presence of 2.5 mM MnCl2. The Km and Vmax of rGaa16Bc for agarose were 6.4 mg/mL and 953 U/mg, respectively. Thin-layer chromatography analysis revealed that rGaa16Bc can hydrolyze agarose into neoagarotetraose and neoagarobiose. Partial hydrolysis products (PHPs) of rGaa16Bc had an average molecular weight of 88–102 kDa and exhibited > 60% hyaluronidase inhibition activity at a concentration of 1 mg/mL, whereas the completely hydrolyzed product (CHP) showed no hyaluronidase at the same concentration. The biochemical properties of Gaa16B suggest that it could be useful for producing functional neoagaro-oligosaccharides. Additionally, the PHP of rGaa16Bc may be useful in promoting its utilization, which is limited due to the gel strength of agar.

scholarly journals A Novel Neoagarotriose-Producing Agarase from the Marine Bacterium Gilvimarinus Agarilyticus JEA5

2020 ◽  
Author(s):  
Youngdeuk Lee ◽  
Eunyoung Jo ◽  
Yeon-Ju Lee ◽  
Hansol Choi ◽  
Tae-Yang Eom ◽  
...  
Keyword(s):  
Marine Bacterium ◽  
Industrial Applications ◽  
Biochemical Properties ◽  
Carbohydrate Binding ◽  
Ionization Mass ◽  
Coding Region ◽  
Protein Activity ◽  
Amino Acid Similarity ◽  
Chromatography Analysis ◽  
Layer Chromatography

Abstract Background The degradation of agar by bacterial agarases has many commercial and academic applications. We recently identified a novel neoagarotriose-producing β-agarase, Gaa16B, in the marine bacterium Gilvimarinus agarilyticus JEA5. This is the first report to describe neoagarotriose production from β-agarase.Results The Gaa16B agarase, which belongs to the glycoside hydrolase 16 (GH16) family of β-agarases, shows less than 70.9% amino acid similarity with previously characterized agarases. The coding region of Gaa16B is 1800 bp long, encoding 600 amino acids, and exhibits features typical of agarases belonging to the GH16 family. A recombinant Gaa16B lacking the carbohydrate binding region (rGaa16Bc) was overexpressed in Escherichia coli and purified as a maltose-binding protein (MBP) fusion protein. Activity assays revealed the optimal temperature and pH of rGaa16Bc to be 55 °C and pH 6–7, respectively, and the protein was highly stable at 55 °C for 90 min. Additionally, rGaa16Bc activity was strongly enhanced (2.3-fold) in the presence of 2.5 mM MnCl2. The Km and Vmax of rGaa16Bc for agarose were 6.4 mg/ml and 953 U/mg, respectively. Thin layer chromatography analysis revealed that rGaa16Bc can hydrolyze agarose into neoagarotetraose, neoagarotriose, and neoagarobiose, and the production of neoagarotriose by rGaa16Bc was successfully validated by high-resolution electrospray ionization mass spectrometry.Conclusion The biochemical properties of Gaa16B and the results of the hydrolytic pattern analysis suggest that Gaa16B could be useful to produce functional neoagaro-oligosaccharides for industrial applications.

scholarly journals The phytochemical screenings and thin layer chromatography analysis of chemical compounds in ethanol extract of labu siam fruit (Sechium edule Jacq. Swartz.)

2005 ◽  
Vol 3 (1) ◽  
pp. 26-31 ◽  
Author(s):  
SOERYA DEWI MARLIANA ◽  
VENTY SURYANTI ◽  
SUYONO SUYONO
Keyword(s):  
Thin Layer ◽  
Petroleum Ether ◽  
Thin Layer Chromatography ◽  
Soxhlet Extraction ◽  
Ethanol Extract ◽  
Chemical Compounds ◽  
Thin Layer Chromatography Analysis ◽  
Chromatography Analysis ◽  
Sechium Edule ◽  
Layer Chromatography

The phytochemical screenings and analysis of chemical compounds in ethanol extract of labu siam fruit (Sechium edule Jacq. Swartz.) with Thin Layer Chromatography (TLC) has been carried out. Isolation was done by Soxhlet extraction for 6 hours with petroleum ether and the residue was extracted by maceration during 24 hours with ethanol.The isolated compounds in ethanol extract were identified by phytochemical screenings method and TLC. The result showed the presence of alkaloid, saponin, cardenolin/bufadienol and flavonoid.

scholarly journals Diagnosis of Oleander Poisoning in Livestock

1996 ◽  
Vol 8 (3) ◽  
pp. 358-364 ◽  
Author(s):  
Francis D. Galey ◽  
Dirk M. Holstege ◽  
Konstanze H. Plumlee ◽  
Elizabeth Tor ◽  
Bill Johnson ◽  
...  
Keyword(s):  
Sudden Death ◽  
Cardiac Arrhythmias ◽  
Clinical Signs ◽  
Laboratory System ◽  
Thin Layer Chromatography Analysis ◽  
Presumptive Diagnosis ◽  
Diagnostic Laboratory ◽  
Chromatography Analysis ◽  
The Past ◽  
Layer Chromatography

Since mid- 1989, 37 cases of oleander poisoning in livestock have been diagnosed at the California Veterinary Diagnostic Laboratory System. The most frequent source for oleander exposure was plant clippings. Sudden death was the most common presenting complaint. Other signs reported included diarrhea, pulmonary edema, tachycardia, cardiac arrhythmias, colic, and lethargy. In the past, a presumptive diagnosis of oleander poisoning could be based only on matching clinical signs with evidence of consumption of oleander. A new 2 dimensional Thin-layer chromatography analysis of ingesta for oleandrin and an awareness of lesions in heart muscle have greatly improved the ability to diagnose oleander toxicosis.

Thin layer chromatography analysis of antioxidant constituents of lichens from Antarctica

2008 ◽  
Vol 62 (4) ◽  
pp. 481-484 ◽  
Author(s):  
Hari Datta Bhattarai ◽  
Babita Paudel ◽  
Soon Gyu Hong ◽  
Hong Kum Lee ◽  
Joung Han Yim
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Thin Layer Chromatography Analysis ◽  
Chromatography Analysis ◽  
Layer Chromatography

Metronidazole uptake in Helicobacter pylori

1995 ◽  
Vol 41 (8) ◽  
pp. 746-749 ◽  
Author(s):  
Richard A. Moore ◽  
Brenda Beckthold ◽  
L. E. Bryan
Keyword(s):  
Helicobacter Pylori ◽  
Resistant Strain ◽  
Sodium Arsenate ◽  
Thin Layer Chromatography Analysis ◽  
Chromatography Analysis ◽  
Metronidazole Resistance ◽  
Carbonyl Cyanide ◽  
H Pylori ◽  
Layer Chromatography ◽  
Imidazole Compounds

Currently, the mechanism of metronidazole resistance is not understood in Helicobacter pylori. We have looked at uptake of metronidazole into a sensitive and a resistant strain of H. pylori. Both strains displayed rapid uptake of [14C]metronidazole, although the resistant strain accumulated the drug at a slower rate and to a lesser amount than the sensitive strain. Uptake was inhibited by KCN and carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) but not by sodium arsenate. Thin-layer chromatography analysis of lysed cell supernatants showed that metronidazole was metabolized in both strains. A variety of related imidazole compounds inhibited metronidazole uptake, consistent with a common transport system for this group of antibiotics. Our data do not support an absence of uptake or metabolism as a cause of resistance in the strain examined.Key words: metronidazole, uptake, Helicobacter.

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