scholarly journals A Novel PL9 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression, and Its Application in Pectin Degradation

2019 ◽  
Vol 20 (12) ◽  
pp. 3060 ◽  
Author(s):  
Ye Yuan ◽  
Xin-Yu Zhang ◽  
Yan Zhao ◽  
Han Zhang ◽  
Yi-Fa Zhou ◽  
...  
Keyword(s):  
High Performance ◽  
Pectate Lyase ◽  
Apparent Molecular Weight ◽  
Paenibacillus Polymyxa ◽  
Single Step ◽  
Water Soluble ◽  
Citrus Pectin ◽  
Pectin Degradation ◽  
Pharmacologically Active

Pectate lyases play an important role in pectin degradation, and therefore are highly useful in the food and textile industries. Here, we report on the cloning of an alkaline pectate lyase gene (pppel9a) from Paenibacillus polymyxa KF-1. The full-length gene (1350 bp) encodes for a 449-residue protein that belongs to the polysaccharide lyase family 9 (PL9). Recombinant PpPel9a produced in Escherichia coli was purified to electrophoretic homogeneity in a single step using Ni2+-NTA affinity chromatography. The enzyme activity of PpPel9a (apparent molecular weight of 45.3 kDa) was found to be optimal at pH 10.0 and 40 °C, with substrate preference for homogalacturonan type (HG) pectins vis-à-vis rhamnogalacturonan-I (RG-I) type pectins. Using HG-type pectins as substrate, PpPel9a showed greater activity with de-esterified HGs. In addition, PpPel9a was active against water-soluble pectins isolated from different plants. Using this lyase, we degraded citrus pectin, purified fractions using Diethylaminoethyl (DEAE)-sepharose column chromatography, and characterized the main fraction MCP-0.3. High-performance gel permeation chromatography (HPGPC) analysis showed that the molecular mass of citrus pectin (~230.2 kDa) was reduced to ~24 kDa upon degradation. Ultra-performance liquid chromatography - tandem mass spectrometer (UPLC-MS) and monosaccharide composition analyses demonstrated that PpPel9a worked as an endo-pectate lyase, which acted primarily on the HG domain of citrus pectin. In vitro testing showed that the degradation product MCP-0.3 significantly promotes the growth of Lactobacillus plantarum and L. rhamnosus. In this regard, the enzyme has potential in the preparation of pharmacologically active pectin products.

scholarly journals Screening of a Novel Polysaccharide Lyase Family 10 Pectate Lyase from Paenibacillus polymyxa KF-1: Cloning, Expression and Characterization

Molecules ◽  
2018 ◽  
Vol 23 (11) ◽  
pp. 2774 ◽  
Author(s):  
Yan Zhao ◽  
Ye Yuan ◽  
Xinyu Zhang ◽  
Yumei Li ◽  
Qiang Li ◽  
...  
Keyword(s):  
Textile Industry ◽  
Pectate Lyase ◽  
Paenibacillus Polymyxa ◽  
Polygalacturonic Acid ◽  
Optimal Conditions ◽  
Citrus Pectin ◽  
Plant Fiber ◽  
Polysaccharide Lyase ◽  
Glycosidic Bonds ◽  
Fiber Processing

Pectate lyase (EC 4.2.2.2) catalyzes the cleavage of α-1,4-glycosidic bonds of pectin polymers, and it has potential uses in the textile industry. In this study, a novel pectate lyase belonging to polysaccharide lyase family 10 was screened from the secreted enzyme extract of Paenibacillus polymyxa KF-1 and identified by liquid chromatography-MS/MS. The gene was cloned from P. polymyxa KF-1 genomic DNA and expressed in Escherichia coli. The recombinant enzyme PpPel10a had a predicted Mr of 45.2 kDa and pI of 9.41. Using polygalacturonic acid (PGA) as substrate, the optimal conditions for PpPel10a reaction were determined to be 50 °C and pH 9.0, respectively. The Km, vmax and kcat values of PpPel10a with PGA as substrate were 0.12 g/L, 289 μmol/min/mg, and 202.3 s−1, respectively. Recombinant PpPel10a degraded citrus pectin, producing unsaturated mono- and oligogalacturonic acids. PpPel10a reduced the viscosity of PGA, and weight loss of ramie (Boehmeria nivea) fibers was observed after treatment with the enzyme alone (22.5%) or the enzyme in combination with alkali (26.3%). This enzyme has potential for use in plant fiber processing.

scholarly journals Novel Antioxidant and Hypoglycemic Water-Soluble Polysaccharides from Jasmine Tea

Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2375
Author(s):  
Yayuan Tang ◽  
Jinfeng Sheng ◽  
Xuemei He ◽  
Jian Sun ◽  
Zhen Wei ◽  
...  
Keyword(s):  
Galacturonic Acid ◽  
High Performance ◽  
Structural Characteristics ◽  
Gel Permeation Chromatography ◽  
Helical Structure ◽  
Water Soluble ◽  
Nuclear Magnetic Resonance Analysis ◽  
Molar Ratios ◽  
Jasmine Tea

There have been few studies dealing with chemical elucidation and pharmacological potentials of water-soluble polysaccharides from jasmine tea, limiting their use in functional foods. In this study, water-soluble polysaccharides (named as JSP) were extracted from Jasminum sambac (L.) Aiton tea and fractionated to afford two sub-fractions (JSP-1 and JSP-2). The main structural characteristics of novel JSP sub-fractions were determined by high performance gel permeation chromatography, ultra-performance liquid chromatography-tandem mass spectrometry, Fourier transform infrared, and nuclear magnetic resonance analysis. Physiologically, the abilities of JSP-1 and JSP-2 to reduce ferric ions, scavenge DPPH and hydroxyl radicals, as well as protect islet cells were confirmed in vitro. JSP-1 exhibited better antioxidant and hypoglycemic activities than JSP-2. The molecular weights of JSP-1 and JSP-2 were 18.4 kDa and 14.1 kDa, respectively. JSP-1 was made up of glucose, galactose, rhamnose, xylose, arabinose, and galacturonic acid with molar ratios 1.14:4.69:1.00:9.92:13.79:4.09, whereas JSP-2 with a triple helical structure was composed of galactose, rhamnose, xylose, arabinose, and galacturonic acid as 3.80:1.00:8.27:11.85:5.05 of molar ratios. JSP-1 contains →1)-α-Galƒ-(3→, →1)-α-Galƒ-(2→, →1)-α-Araƒ-(5→, →1)-α-Araƒ-(3→, →1)-α-Araƒ-(3,5→, →1)-β-Xylp-(2→ and →1)-β-Xylp-(3→ residues in the backbone. These results open up new pharmacological prospects for the water-soluble polysaccharides extracted from jasmine tea.

scholarly journals Development of Chitosan–Tripolyphosphate Nanoparticles as Glycopeptide Antibiotic Reservoirs and Ex vivo Evaluation for Their Potential to Enhance the Corneal Permeation in Ocular Drug Delivery

2021 ◽  
Author(s):  
Farhad Safari ◽  
Shahla Mirzaeei ◽  
Ghobad Mohammadi
Keyword(s):  
Ftir Spectroscopy ◽  
High Performance ◽  
Ex Vivo ◽  
Antimicrobial Efficacy ◽  
Ocular Delivery ◽  
In Vivo Evaluation ◽  
Pharmacologically Active ◽  
Corneal Permeation

Purpose: The present investigation aimed to prepare Vancomycin-loaded nanoparticles (VAN-NPs) using chitosan (CS) and tripolyphosphate (TPP) besides exploring the effects of changing CS/TPP ratio on the physicochemical properties, corneal permeation, and ocular delivery of the prepared NPs. Methods: Different pre-formulations were prepared using the modified ionic gelation process, then were characterized in terms of size distribution. Optimized formulations were furtherly evaluated by some characteristic tools such as Fourier-transform infrared (FTIR) spectroscopy and thermogravimetric analysis (TGA). The in vitro antimicrobial efficacy and drug release amounts along with the Ex-vivo corneal permeation of NPs through the sheep cornea were investigated. Quantification was performed using High-Performance Liquid Chromatography. Results: Spherical and uniformly distributed NPs were developed with a mean particle size varied between 215–290 nm. FTIR spectroscopy confirmed that the CS/TPP cross-linking has taken place without affecting the pharmacologically active moiety of the drug. The obtained zeta potential values were in the range of +34 to +37 mV, which could ensure the stability of formulations. TGA analysis indicated enhanced thermal stability for the encapsulated drug compared to the plain drug. Formulations indicated suitable antimicrobial efficacy while releasing more than 90% of the drug during 24 h. NPs offered a 10-fold enhancement in corneal permeation compared to the drug solution. Conclusions: Although further in vivo evaluation is still required to completely confirm the efficacy of the formulations, the enhanced release and corneal permeation of the drug suggest that the prepared NPs are suitable for ocular delivery of VAN.

scholarly journals Pharmacokinetics of Amino Acid Phosphoramidate Monoesters of Zidovudine in Rats

2002 ◽  
Vol 46 (5) ◽  
pp. 1357-1363 ◽  
Author(s):  
Heng Song ◽  
George W. Griesgraber ◽  
Carston R. Wagner ◽  
Cheryl L. Zimmerman
Keyword(s):  
Amino Acid ◽  
High Performance ◽  
Volume Of Distribution ◽  
Water Soluble ◽  
Pharmacokinetic Parameters ◽  
Sprague Dawley ◽  
Binding Studies ◽  
Total Body

ABSTRACT In vitro studies have demonstrated that water-soluble, nontoxic phosphoramidates of azidothymidine (zidovudine [AZT]) have significant and specific anti-human immunodeficiency virus and anticancer activity. Although polar, these compounds are internalized and processed to the corresponding nucleoside monophosphates. Eight methyl amide and methyl ester phosphoramidate monoesters composed of d- or l-phenylalanine or tryptophan and AZT were synthesized. The plasma stability and protein binding studies were carried out in vitro. Then in vivo pharmacokinetic evaluations of six of the compounds were conducted. Sprague-Dawley rats received each compound by intravenous bolus dose, and serial blood and urine samples were collected. AZT and phosphoramidate concentrations in plasma and urine were quantitated by high-performance liquid chromatography with UV or fluorescence detection. Pharmacokinetic parameters were calculated by standard noncompartmental means. The plasma half-lives of the phosphoramidates were 10- to 20-fold longer than the half-life of AZT. Although the renal clearances of the phosphoramidates were similar to AZT, their total body clearances were significantly greater than that of AZT. The 3- to 15-fold-larger volume of distribution (V ss) for the phosphoramidates relative to AZT appeared to be dependent on the stereochemistry of the amino acid, with the largest values being associated with the l-amino acids. The increased V ss indicates a much greater tissue distribution of the phosphoramidate prodrugs than of AZT. Amino acid phosphoramidate monoesters of AZT have improved pharmacokinetic properties over AZT and significant potential as in vivo pronucleotides.

scholarly journals Structural Characterization and Antioxidative Activity of Low-Molecular-Weights Beta-1,3-Glucan from the Residue of ExtractedGanoderma lucidumFruiting Bodies

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Pai-Feng Kao ◽  
Shwu-Huey Wang ◽  
Wei-Ting Hung ◽  
Yu-Han Liao ◽  
Chun-Mao Lin ◽  
...  
Keyword(s):  
Cell Death ◽  
High Performance ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Water Soluble ◽  
Dependent Manner ◽  
Ros Production ◽  
Fruiting Bodies ◽  
Concentration Dependent Manner

The major cell wall constituent ofGanoderma lucidum(G. lucidum) isβ-1,3-glucan. This study examined the polysaccharide from the residues of alkaline-extracted fruiting bodies using high-performance anion-exchange chromatography (HPAEC), and it employed nuclear magnetic resonance (NMR) and mass spectrometry (MS) to confirm the structures. We have successfully isolated low-molecular-weightβ-1,3-glucan (LMG), in high yields, from the waste residue of extracted fruiting bodies ofG. lucidum. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay evaluated the capability of LMG to suppress H2O2-induced cell death in RAW264.7 cells, identifying that LMG protected cells from H2O2-induced damage. LMG treatment decreased H2O2-induced intracellular reactive oxygen species (ROS) production. LMG also influenced sphingomyelinase (SMase) activity, stimulated by cell death to induce ceramide formation, and then increase cell ROS production. Estimation of the activities of neutral and acid SMasesin vitroshowed that LMG suppressed the activities of both neutral and acid SMases in a concentration-dependent manner. These results suggest that LMG, a water-solubleβ-1,3-glucan recycled from extracted residue ofG. lucidum, possesses antioxidant capability against H2O2-induced cell death by attenuating intracellular ROS and inhibiting SMase activity.

scholarly journals A Class of Pantothenic Acid Analogs Inhibits Plasmodium falciparum Pantothenate Kinase and Represses the Proliferation of Malaria Parasites

2005 ◽  
Vol 49 (11) ◽  
pp. 4649-4657 ◽  
Author(s):  
Christina Spry ◽  
Christina L. L. Chai ◽  
Kiaran Kirk ◽  
Kevin J. Saliba
Keyword(s):  
Plasmodium Falciparum ◽  
Cell Line ◽  
Pantothenic Acid ◽  
Human Cell Line ◽  
Single Step ◽  
Water Soluble ◽  
Pantothenate Kinase ◽  
Absolute Requirement ◽  
Essential Enzyme

ABSTRACT The growth and proliferation of the human malaria parasite Plasmodium falciparum are dependent on the parasite's ability to obtain essential nutrients. One nutrient for which the parasite has an absolute requirement is the water-soluble vitamin pantothenic acid (vitamin B5). In this study, a series of pantothenic acid analogs which retain the 2,4-dihydroxy-3,3-dimethylbutyramide core of pantothenic acid but deviate in structure from one another and from pantothenic acid in the nature of the substituent attached to the amide nitrogen were synthesized using an efficient single-step synthetic route. Eight of 10 analogs tested inhibited the proliferation of intraerythrocytic P. falciparum parasites in vitro, doing so with 50% inhibitory concentrations between 15 and 200 μM. The compounds were generally selective, inhibiting the proliferation of a human cell line (the Jurkat cell line) only at concentrations severalfold higher than those required for inhibition of parasite growth. It was demonstrated that compounds in this series inhibited the phosphorylation of pantothenic acid by pantothenate kinase, the first step in the parasite's biosynthesis of the essential enzyme cofactor coenzyme A, doing so competitively, with Ki values in the nanomolar range.

scholarly journals In vitro Assessment of Chemical and Pre-biotic Properties of Carboxymethylated Polysaccharides From Passiflora edulis Peel, Xylan, and Citrus Pectin

2021 ◽  
Vol 8 ◽  
Author(s):  
Yongjin Sun ◽  
Yuan Guan ◽  
Hock Eng Khoo ◽  
Xia Li
Keyword(s):  
Culture Media ◽  
Chloroacetic Acid ◽  
Streptococcus Thermophilus ◽  
Lactobacillus Brevis ◽  
Water Soluble ◽  
Lactobacillus Delbrueckii ◽  
Citrus Pectin ◽  
Passiflora Edulis ◽  
Probiotic Strains

This study aimed to determine the carboxymethylation effect of crude water-soluble polysaccharides of Passiflora edulis peel (WPEP), xylan (XY), and citrus pectin (CP). Their chemical and pre-biotic properties were also determined. The polysaccharides were carboxymethylated by reacting with chloroacetic acid and sodium hydroxide. The carboxymethylated and non-carboxymethylated polysaccharides were also used as pre-biotics to study the growth pattern of selected intestinal microflora. These polysaccharides substituted the glucose solution in culture media for culturing Lactobacillus brevis GIM1.773, Lactobacillus plantarum GIM1.19, Lactobacillus delbrueckii subsp. bulgaricus GIM1.155, and Streptococcus thermophilus GIM1.540. The results showed that the carboxymethylated polysaccharides c-XY, c-CP, and c-WPEP, had substitution degrees of 0.682, 0.437, and 0.439, respectively. The polysaccharides demonstrated resistance to digestion in the simulated human digestive models. The resistance to digestion was enhanced by carboxymethylation, especially the carboxymethylated CP and WPEP. The results also showed that the pre-biotic activities of the polysaccharides increased after carboxymethylation. The c-XY had a better pre-biotic effect than XY and the other carbohydrate samples. The findings suggested that carboxymethylated polysaccharides may be developed into novel pre-biotics and nutraceuticals that could promote growth of the probiotic strains.

scholarly journals Water-Soluble Melanoidin Pigment as a New Antioxidant Component of Fermented Willowherb Leaves (Epilobium angustifolium)

Antioxidants ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1300
Author(s):  
Daniil N. Olennikov ◽  
Christina S. Kirillina ◽  
Nadezhda K. Chirikova
Keyword(s):  
High Performance ◽  
Chemical Shifts ◽  
Water Soluble ◽  
Antioxidant Potential ◽  
Photodiode Array Detection ◽  
Plant Materials ◽  
Isolation Technique ◽  
Epilobium Angustifolium ◽  
General Chemical

Willowherb (Epilobium angustifolium L., family Onagraceae) is a well-known food and medicinal plant used after fermentation as a source of beverages with high antioxidant potential. Despite this long history of use, only a few papers have described the chemical profile and bioactivity of fermented willowherb tea in general. To understand the basic metabolic differences of non-fermented and fermented E. angustifolium leaves, we used general chemical analysis, high-performance liquid chromatography with photodiode array detection and electrospray ionization triple quadrupole mass spectrometric detection assay, and an isolation technique. As a result, the content of 14 chemical groups of compounds was compared in the two plant materials; 59 compounds were detected, including 36 new metabolites; and a new water-soluble phenolic polymer of melanoidin nature was isolated and characterized. The fundamental chemical shifts in fermented E. angustifolium leaves relate mainly to the decrease of ellagitannin content, while there is an increase of melanoidin percentage and saving of the antioxidant potential, despite the significant changes detected. The strong antioxidative properties of the new melanoidin were revealed in a series of in vitro bioassays, and a simulated gastrointestinal and colonic digestion model demonstrated the stability of melanoidin and its antioxidant activity. Finally, we concluded that the new melanoidin is a basic antioxidant of the fermented leaves of E. angustifolium, and it can be recommended for additional study as a promising food and medicinal antioxidant agent.

Radiation Protection of Carboxymethylation of an Acid Polysaccharides Extracted from Auricularia Auricula against Uvb In Vitro

2011 ◽  
Vol 365 ◽  
pp. 199-204
Author(s):  
Hua Zhang ◽  
Zhen Yu Wang ◽  
Zi Luan Fan ◽  
Xin Yang ◽  
Xue Wang ◽  
...  
Keyword(s):  
Radiation Protection ◽  
High Performance ◽  
Dose Range ◽  
Chemical Characterization ◽  
Water Soluble ◽  
Molar Ratio ◽  
Ft Ir ◽  
Acid Polysaccharide ◽  
Soluble Acid

This paper reports on a water-soluble acid polysaccharide (AAP) and in how it was extracted from Auricularia auricular, acquired by CTAB, and prepared it’s carboxymethylation. Chemical characterization by high-performance liquid chromatography/gel permeation chromatograph (HPLC/GPC), Fourier transform infrared (FT-IR) spectrometer and gas chromatography–mass spectrophotometer (GC–MS) were investigated. Chemical analysis indicated that C AAAP was composed of arabinose, xylose, mannose, glucose and galactose, with the molar ratio at 0.04: 0.13: 1.00: 0.59: 0.29. Moreover, radiation protection against UVB in vitro indicated that at the dose range of 200–500 μg/mL, C AAAP enhanced the protection of HepG2 cells against UVB cytotoxicity than AAAP. However, but at the dose range of 50–150 μg/mL the result was just opposite.

scholarly journals Investigation of In Vivo and In Vitro Pharmacokinetic Characteristics of Kaji-ichigoside F1 in Rats

2020 ◽  
Vol 15 (3) ◽  
pp. 1934578X2091501
Author(s):  
Wenjuan Cui ◽  
Xiaoguang Fan ◽  
Naizhi Wang ◽  
Zhikun Zhang ◽  
Zhaolong Zhang ◽  
...  
Keyword(s):  
High Performance ◽  
Single Step ◽  
Selected Reaction Monitoring ◽  
Pharmacokinetic Studies ◽  
Triterpenoid Saponin ◽  
Pentacyclic Triterpenoid ◽  
Validation Parameters ◽  
Liquid Chromatography Tandem Mass

Kaji-ichigoside F1, a pentacyclic triterpenoid saponin, exhibits various beneficial pharmacological effects. In this study, a simple, rapid, and specific high-performance liquid chromatography-tandem mass spectrometry method for rapid quantification of kaji-ichigoside F1 in rat biological matrix was developed. Plasma was prepared by a single-step protein precipitation followed by separation of the analyte using an Inertsil ODS-3 column with a gradient mobile phase. Positive ion electrospray was used and selected reaction monitoring transitions were m/ z 673.27 → 511.15 for kaji-ichigoside F1 and m/ z 429.19 → 267.29 for morroniside, respectively. The developed method was validated with linear range of 20 to 10 000 ng/mL. All validation parameters were well within the acceptance limit based on the guidance of FDA. The validated method was successfully applied to analyze samples from the in vivo and in vitro pharmacokinetic studies in rats.

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