scholarly journals Identification of aSaccharomyces cerevisiaeGlucosidase That Hydrolyzes Flavonoid Glucosides

2011 ◽  
Vol 77 (5) ◽  
pp. 1751-1757 ◽  
Author(s):  
Sabine Schmidt ◽  
Sandra Rainieri ◽  
Simone Witte ◽  
Ulrich Matern ◽  
Stefan Martens
Keyword(s):  
High Performance ◽  
Hydrolytic Activity ◽  
Glycoside Hydrolases ◽  
Yeast Cells ◽  
Genome Database ◽  
Yeast Transformants ◽  
Product Verification ◽  
Layer Chromatography ◽  
Hydrolysis Of

ABSTRACTBaker's yeast (Saccharomyces cerevisiae) whole-cell bioconversions of naringenin 7-O-β-glucoside revealed considerable β-glucosidase activity, which impairs any strategy to generate or modify flavonoid glucosides in yeast transformants. Up to 10 putative glycoside hydrolases annotated in theS. cerevisiaegenome database were overexpressed with His tags in yeast cells. Examination of these recombinant, partially purified polypeptides for hydrolytic activity with synthetic chromogenic α- or β-glucosides identified three efficient β-glucosidases (EXG1, SPR1, and YIR007W), which were further assayed with natural flavonoid β-glucoside substrates and product verification by thin-layer chromatography (TLC) or high-performance liquid chromatography (HPLC). Preferential hydrolysis of 7- or 4′-O-glucosides of isoflavones, flavonols, flavones, and flavanones was observedin vitrowith all three glucosidases, while anthocyanins were also accepted as substrates. The glucosidase activities of EXG1 and SPR1 were completely abolished by Val168Tyr mutation, which confirmed the relevance of this residue, as reported for other glucosidases. Most importantly, biotransformation experiments with knockout yeast strains revealed that only EXG1 knockout strains lost the capability to hydrolyze flavonoid glucosides.

Phytochemical, Analytical and Medicinal Studies of Holoptelea integrifolia Roxb. Planch - A Review

2019 ◽  
Vol 5 (4) ◽  
pp. 270-277 ◽  
Author(s):  
Vijay Kumar ◽  
Simranjeet Singh ◽  
Ragini Bhadouria ◽  
Ravindra Singh ◽  
Om Prakash
Keyword(s):  
Liquid Chromatography ◽  
Thin Layer ◽  
Thin Layer Chromatography ◽  
High Performance ◽  
Biologically Active ◽  
Toxicological Studies ◽  
Wide Range ◽  
Layer Chromatography

Holoptelea integrifolia Roxb. Planch (HI) has been used to treat various ailments including obesity, osteoarthritis, arthritis, inflammation, anemia, diabetes etc. To review the major phytochemicals and medicinal properties of HI, exhaustive bibliographic research was designed by means of various scientific search engines and databases. Only 12 phytochemicals have been reported including biologically active compounds like betulin, betulinic acid, epifriedlin, octacosanol, Friedlin, Holoptelin-A and Holoptelin-B. Analytical methods including the Thin Layer Chromatography (TLC), High-Performance Thin Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC) and Liquid Chromatography With Mass Spectral (LC-MS) analysis have been used to analyze the HI. From medicinal potency point of view, these phytochemicals have a wide range of pharmacological activities such as antioxidant, antibacterial, anti-inflammatory, and anti-tumor. In the current review, it has been noticed that the mechanism of action of HI with biomolecules has not been fully explored. Pharmacology and toxicological studies are very few. This seems a huge literature gap to be fulfilled through the detailed in-vivo and in-vitro studies.

scholarly journals Establishment of anIn Vitrod-Cycloserine-Synthesizing System by UsingO-Ureido-l-Serine Synthase and d-Cycloserine Synthetase Found in the Biosynthetic Pathway

2013 ◽  
Vol 57 (6) ◽  
pp. 2603-2612 ◽  
Author(s):  
Narutoshi Uda ◽  
Yasuyuki Matoba ◽  
Takanori Kumagai ◽  
Kosuke Oda ◽  
Masafumi Noda ◽  
...  
Keyword(s):  
Gene Cluster ◽  
High Performance ◽  
Sequence Similarity ◽  
Open Reading Frames ◽  
Homocysteine Thiolactone ◽  
Putative Gene ◽  
Content Type ◽  
Dna Fragment ◽  
Layer Chromatography

ABSTRACTWe have recently cloned a DNA fragment containing a gene cluster that is responsible for the biosynthesis of an antituberculosis antibiotic,d-cycloserine. The gene cluster is composed of 10 open reading frames, designateddcsAtodcsJ. Judging from the sequence similarity between each putative gene product and known proteins, DcsC, which displays high homology to diaminopimelate epimerase, may catalyze the racemization ofO-ureidoserine. DcsD is similar toO-acetylserine sulfhydrylase, which generatesl-cysteine usingO-acetyl-l-serine with sulfide, and therefore, DcsD may be a synthase to generateO-ureido-l-serine usingO-acetyl-l-serine and hydroxyurea. DcsG, which exhibits similarity to a family of enzymes with an ATP-grasp fold, may be an ATP-dependent synthetase convertingO-ureido-d-serine intod-cycloserine. In the present study, to characterize the enzymatic functions of DcsC, DcsD, and DcsG, each protein was overexpressed inEscherichia coliand purified to near homogeneity. The biochemical function of each of the reactions catalyzed by these three proteins was verified by thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and, in some cases, mass spectrometry. The results from this study demonstrate that by using a mixture of the three purified enzymes and the two commercially available substratesO-acetyl-l-serine and hydroxyurea, synthesis ofd-cycloserine was successfully attained. Thesein vitrostudies yield the conclusion that DcsD and DcsG are necessary for the syntheses ofO-ureido-l-serine andd-cycloserine, respectively. DcsD was also able to catalyze the synthesis ofl-cysteine when sulfide was added instead of hydroxyurea. Furthermore, the present study shows that DcsG can also form other cyclicd-amino acid analogs, such asd-homocysteine thiolactone.

scholarly journals HPTLC METHOD FOR ESTIMATION AND QUANTIFICATION OF β-SITOSTEROL FROM IN-VIVO AND IN-VITRO SAMPLES OF MERREMIA AEGYPTIA AND MERREMIA DISSECTA

2020 ◽  
pp. 162-165
Author(s):  
RIDHI JOSHI ◽  
RISHIKESH MEENA ◽  
PREETI MISHRA ◽  
VIDYA PATNI
Keyword(s):  
High Performance ◽  
Normal Phase ◽  
Leaf Sample ◽  
Plant Parts ◽  
Methanolic Extracts ◽  
Aluminum Plates ◽  
Layer Chromatography ◽  
Hptlc Method

Objective: A normal-phase high-performance thin-layer chromatography (HPTLC) method has been developed and validated for estimation and quantitation of beta-sitosterol from the methanolic fraction of different plant parts of two medicinally important plants viz. Merremia aegyptia and Merremia dissecta. These plants have been reported to possess antimicrobial, antioxidant, and anti-inflammatory activities. Methods: Chromatographic separation of beta-sitosterol from the methanolic extracts of plant parts of M. aegyptia and M. dissecta was performed on TLC aluminum plates pre-coated with silica gel 60F254 using a suitable mobile phase. The densitometric scanning was done after derivatization at ????-580 nm for ????-sitosterol. Result: Only M. dissecta leaf sample was reported to contain ????-sitosterol (4.6 ng/μl), whereas other samples such as seed, stem, and callus extracts of M. aegyptia and M. dissecta did not showed its presence. Conclusion: The developed HPTLC method is simple, rapid, and precise and can be used for routine analysis and quantification of ????-sitosterol and other useful plant bioactives that are phytopharmaceutically important.

In vitro digestibility study of thermal oxidized palm oleins Estudio de la digestibilidad in vitro de oleínas de palma termooxidadas

2000 ◽  
Vol 6 (6) ◽  
pp. 449-456 ◽  
Author(s):  
F.J. Sánchez-Muniz ◽  
R. Arroyo ◽  
J.M. Sánchez-Montero ◽  
C. Cuesta
Keyword(s):  
Pancreatic Lipase ◽  
High Performance ◽  
Palm Olein ◽  
Size Exclusion ◽  
In Vitro Digestibility ◽  
Moderate Degree ◽  
Before And After ◽  
Exclusion Chromatography ◽  
Hydrolysis Of

Information on digestibility and absorption of oils and fats used for frying is under debate. To get knowledge on this, unused palm olein (9.27 ± 0.10% w/w polar content), used frying palm olein with a moderate degree of alteration (14.81 ± 0.90% w/w polar content) and highly altered used frying palm olein (26.36 ± 0.30% w/w polar content) and their respective nonpolar and polar fractions were studied. Samples were analyzed by high-performance size-exclusion chromatography before and after a 20-min in vitro incubation with pancreatic lipase. Formation of monoacylglycerols and free fatty acids reflected no relevant differences between unused and moderately altered oleins, whereas the most altered olein was hydrolyzed to a much lesser degree. The presence of oligomers (dimers and polymers of triacylglycerols) negatively affected the hydrolysis of triacylglycerol monomers in whole oleins. The hydrolysis of these monomers in the isolated nonpolar and polar fractions ranked between 60.2% and 78.5%. Oligomers were efficiently hydrolyzed by pancreatic lipase in whole un used and moderately altered oleins but not in the most altered one. Polymers from isolated polar fractions were poorly hydrolyzed or not hydrolyzed at all. These data suggest that whole oleins contained some compounds that increase susceptibility of oligomers to enzymatic hydrolysis and that such compounds were not present in the polar fraction.

scholarly journals THE PRELIMINARY STUDY OF ANTIOXIDANT ACTIVITY FROM XYLO-OLIGOSACCHARIDE OF CORNCOB (ZEA MAYS) HYDROLYSIS PRODUCT WITH ENDO-β-XYLANASE ENZYME

2016 ◽  
Vol 3 (2) ◽  
pp. 112 ◽  
Author(s):  
Laura Navika Yamani ◽  
Alfinda Novi Kristanti ◽  
Ni Nyoman Puspaningsih
Keyword(s):  
Antioxidant Activity ◽  
High Performance ◽  
Hot Spring ◽  
Hydrolysis Product ◽  
Xylanase Enzyme ◽  
Preliminary Study ◽  
Layer Chromatography ◽  
Anti Tumor Activity ◽  
Oligosaccharide Hydrolysis

Xylo-oligosaccharide derived from corncob hemicellulose has been reported to possess antioxidant activity. In order to assess the effective scavenging of xylo-oligosaccharide, we conducted in vitro studies based on self-made xylo-oligosaccharide with DPPH (2,2diphenyl-1-picrilhydrazil) method. Xylo-oligosaccharide was prepared with enzymatic hydrolysis. The enzyme used for hemicellulose hydrolysis was endo-β-xylanase enzyme from PC-01 isolated bactrerium. PC-01 isolated bacterium used in this study was Pacet hot spring which was isolated from East Java. Endo-β-xylanase enzyme is an extracelluler enzyme. There was about 0.199 U/mL after purification and dialysis process. Hydrolisis product of hemicellulose A and B from corncob were analyzed with TLC (Thin Layer Chromatography) and HPLC (High Performance Liquid Chromatography). This analysis showed that hydrolysis product of hemicellulose B had a lot of xylo-oligosaccharide hydrolysis product of hemicellulose than Xylo-oligosaccharide hydrolysis product of hemicelluloses A. Xylo-olygosaccharide was analyzed as on antioxidant activity. Xylo-oligosaccharide hydrolysis product ofhemicellulose B (IC = 48.96) has higher antioxidant activity than Xylo-oligosaccharide hydrolysis product of hemicellulose A (IC 50 50 = 92.302). The toxicity of xylo-oligosaccharide can be calculated by the value of LC 50 (Lethality concentration). LC of xylooligosaccharide derived from corncob hemicellulose was 400 ppm so that xylo-oligosaccharide has anti tumor activity because xylooligosaccharide has LC 50 < 1000 ppm.

scholarly journals High performance thin-layer chromatography and in vitro cytotoxic studies on ethanol extract of Matricaria chamomilla L (Asteraceae) flowers

2021 ◽  
Vol 18 (9) ◽  
pp. 1969-1976
Author(s):  
Mohammed Al Bratty ◽  
Lalitha K. Govindaram ◽  
Lalitha K. Govindaram ◽  
Neelaveni Thangavel ◽  
Hassan A. Alhazmi ◽  
...  
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Mtt Assay ◽  
High Performance ◽  
Cytotoxic Effect ◽  
Ethanol Extract ◽  
Matricaria Chamomilla ◽  
Layer Chromatography ◽  
Mcf 7

Purpose: To develop a high performance thin-layer chromatography (HPTLC procedure for quantitation of apigenin in ethanol extract of Matricaria chamomilla (Babunaj) flowers, and to evaluate the extract for in vitro cytotoxic effect on MCF-7 cell lines. Methods: Quantification of apigenin was carried out using a CAMAG TLC system. A combination of toluene, ethyl acetate and formic acid (4.5:3.5:0.2 v/v/v) was used as mobile phase, with densitometry detection at 336 nm. The HPTLC procedure was subjected to validation as per ICH guidelines. The cytotoxicity of the extract was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Results: A sharp apigenin band at Rf of 0.51 was obtained, and the content of apigenin in the extract was 0.062 % w/w. The detection limit (LOD) and quantification limit (LOQ) were 0.19 and 0.57 ng/band, respectively. MTT assay results indicate that M. chamomilla was cytotoxic to Michigan Cancer Foundation-7 (MCF-7) cells, with half-maximal concentration (IC50) of 74 µg/mL. Conclusion: The developed HPTLC method is linear, precise, accurate and specific for the determination of apigenin. M. chamomilla exerts cytotoxic effect on MCF-7 cell line via induction of apoptosis.

scholarly journals Determination of flavones in species of Thymus L. (Lamiaceae) from Macedonian flora

2001 ◽  
Vol 47 ◽  
pp. 9-14
Author(s):  
Svetlana Kulevanova ◽  
Marina Stefova ◽  
Tatjana Kadifkova Panovska ◽  
Jasmina Tonic ◽  
Trajce Stafilov
Keyword(s):  
Ethyl Acetate ◽  
High Performance ◽  
Gradient Elution ◽  
Hplc Method ◽  
Total Flavonoids ◽  
Column Temperature ◽  
Plant Samples ◽  
Layer Chromatography ◽  
Hydrolysis Of

Assay of flavonoids in extracts of seven Thymus L. (Lamiaceae) species from Macedonia including identification and quantification was performed. Extracts obtained after hydrolysis of air dried samples (A1) were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). Luteolin and apigenin were identified in comparison to authentic standard substances. The content of total flavonoids in plant samples determined by UV-Vis spectrometry (with AlCl3) ranged from 0.05-0.13 %. Two other extracts were prepared by extraction with a mixture of ethanol:water (7:3, V/V), evaporation until only water remained and extraction first with diethylether (A2) and secondly with ethyl acetate (A3). The content of flavonoids in diethyl-ether and ethyl acetate extracts ranged from 52.5-244.4 mg·ml-1 and 48.7 -117.5 mg·ml-1, respectively. For quantification of luteolin and total flavonoids the HPLC method was applied, using reverse phase column C18, mobile phase consisting of 5% acetic acid and methanol in gradient elution mode and column temperature set to 40 o C. The content of luteolin in the plant samples ranged from 0.23-0.48 % (m/m), while the content of total flavonoids was found to be 0.26-0.52 %.

scholarly journals Phytochemical characterization of the ethanolic extract of Kaempferia galanga rhizome for anti-oxidant activities by HPTLC and GCMS

2022 ◽  
Vol 8 (1) ◽  
Author(s):  
Flavius Phrangsngi Nonglang ◽  
Abhijeet Khale ◽  
Surya Bhan
Keyword(s):  
Gas Chromatography ◽  
High Performance ◽  
Freeze Drying ◽  
Ethanolic Extract ◽  
Dry Weight ◽  
Kaempferia Galanga ◽  
Anti Oxidant ◽  
Gas Chromatography Mass Spectroscopy ◽  
Layer Chromatography

Abstract Background The rhizome of Kaempferia galanga (K. galanga) was collected from Meghalaya, India, and its ethanolic extract was obtained by freeze-drying or lyophilization process, which was then assessed for its in vitro anti-oxidant activity and phytochemical characterization using high-performance thin-layer chromatography (HPTLC) and gas chromatography-mass spectroscopy (GCMS). Results In vitro anti-oxidant activity analysis shows an inhibitory concentration (IC50) value of 1.824 mg/mL and 0.307 mg/mL for, α, α-diphenyl-ρ-picrylhydrazyl (DPPH) and 2, 2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays, respectively. Total polyphenol content (TPC) of 23.55 ± 0.5 mg gallic acid equivalent (GAE)/g dry weight of extract and total flavonoid content (TFC) of 100 ± 1.414 mg rutin equivalents (RE)/g dry weight of extract were found. High-performance thin-layer chromatography (HPTLC) analysis shows the best separation of bands at different retention factor (Rf) values, when employing the solvent system 2-butanol/1-propanol/water in the ratio of 3:1:1 (v/v/v). Gas chromatography-mass spectroscopy (GCMS) analysis confirms the presence and identification of various phytocompounds, with ethyl p-methoxycinnamate identified as the major active compound. Conclusion Freeze-dried ethanolic extract of K. galanga (rhizome) possesses anti-oxidant activity. Ethyl p-methoxycinnamate is present as the major bioactive component (about 94.87% of the total area composition), and since it has very important and diverse medicinal properties, a freeze-drying process (lyophilization) can be utilized for its isolation and extraction.

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