Column Chromatographic Method Research Articles

A sustainable bioprocess technology for producing food-flavour (+)-γ-decalactone from castor oil-derived ricinoleic acid using enzymatic activity of Candida parapsilosis: Scale-up optimization and purification using novel composite

Ricinoleic acid (RA) from castor oil was employed in biotransformation of peach-flavoured γ-decalactone (GDL), using a Candida parapsilosis strain (MTCC13027) which was isolated from waste of pineapple crown base. Using four variables—pH, cell density, amount of RA, and temperature—the biotransformation parameters were optimized using RSM and BBD. Under optimized conditions (pH 6, 10 % of microbial cells, 10 g/L RA at 28°C), the conversion was maximum and resulted to 80 % (+)-GDL (4.4 g/L/120 h) yield in shake flask (500 mL). Furthermore, optimization was achieved by adjusting the aeration and agitation parameters in a 3 L bioreactor, which were then replicated in a 10 L bioreactor to accurately determine the amount of (+)-GDL. In bioreactor condition, 4.7 g/L (>85 %) of (+)-GDL is produced with 20 % and 40 % dissolved oxygen (1.0 vvm) at 150 rpm in 72 h and 66 h, respectively. Further, a new Al-Mg-Ca-Si composite column-chromatography method is developed to purify enantiospecific (+)-GDL (99.9 %). This (+)-GDL is 100 % nature-identical as validated through 14C-radio-carbon dating. Thorough chemical investigation of enantiospecific (+)-GDL is authenticated for its use as flavour. This bioflavour has been developed through a cost-effective biotechnological process in response to the demand from the food industry on commercial scale.

High-purity 1,2-dimyristoyl-sn-glycero-3-phosphocholine: synthesis and emulsifying performance evaluation.

1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC) is a promising emulsifier for bioactive delivery systems, but its industrial applications are limited by the lack of cost-effective and scalable synthetic routes. The purpose of this study was to economically produce high-purity DMPC by replacing commonly used column chromatography methods and to evaluate the emulsifying performance. DMPC was synthesized from sn-glycero-3-phosphocholine using Steglich esterification followed by sequential recrystallization from ethyl acetate and acetone. The structure of DMPC was identified and its purity was confirmed using various spectroscopy and chromatography techniques. The emulsifying performance was evaluated by examining the effects of storage on the properties of o/w emulsions prepared using soybean oil with (i) soy PC, (ii) soy PC + DMPC (1:1, w/w), and (iii) DMPC as emulsifiers. The chemical impurities formed during the synthesis of DMPC was removed, and its final purity was 96%, and the melt transition temperature was 37.6°C. No visible difference between the three emulsions (soy PC, soy PC+DMPC, and DMPC) was observed during two-week storage, and the DMPC-based emulsion was more stable than soy PC emulsion, showing smaller particle size distribution during 6 months. The highly pure DMPC was synthesized by an economical method, and DMPC-based emulsions demonstrated physicochemical stable, highlighting its potential for food and pharmaceutical industry-related applications. Our findings suggest that DMPC holds promise as an emulsifier with broad applications in the food industry.

Antimicrobial, Antioxidant, and Antityrosinase Activities of Morina persica L. and Its Isolated Compounds.

In this study, the isolation of compounds from the aerial parts of Morina persica L. and the antimicrobial, antioxidant and antityrosinase activities of various polarity extracts and isolated compounds were investigated. Column chromatography methods were used for isolation. A microdilution method was used to determine antimicrobial activity; Folin-Ciocalteu method was used to determine total phenolic content; DPPH and ABTS radical scavenging- capacity methods were used to determine antioxidant activity; and a mushroom tyrosinase method was used to determine antityrosinase activity. Kaempferol-3-O-β-glucopyranoside (astragalin) and quercetin-3-O-rutinoside (rutin) were isolated from M. persica. The extracts and compounds showed higher activity against Staphylococcus aureus and Enterococcus faecalis than other tested bacteria. The highest phenolic content, DPPH, and ABTS radical scavenging activity were detected in an ethyl acetate extract at 50 μg/mL concentration. The methanol extract showed the highest antityrosinase effect at 200 μg/mL concentration.

Dicerandrol C Suppresses Proliferation and Induces Apoptosis of HepG2 and Hela Cancer Cells by Inhibiting Wnt/β-Catenin Signaling Pathway.

Overwhelming evidence points to an aberrant Wnt/β-catenin signaling as a critical factor in hepatocellular carcinoma (HCC) and cervical cancer (CC) pathogenesis. Dicerandrol C (DD-9), a dimeric tetrahydroxanthenone isolated from the endophytic fungus Phomopsis asparagi DHS-48 obtained from mangrove plant Rhizophora mangle via chemical epigenetic manipulation of the culture, has demonstrated effective anti-tumor properties, with an obscure action mechanism. The objective of the current study was to explore the efficacy of DD-9 on HepG2 and HeLa cancer cells and its functional mechanism amid the Wnt/β catenin signaling cascade. Isolation of DD-9 was carried out using various column chromatographic methods, and its structure was elucidated with 1D NMR. The cytotoxicity of DD-9 on HepG2 and HeLa cells was observed with respect to the proliferation, clonality, migration, invasion, apoptosis, cell cycle, and Wnt/β-catenin signaling cascade. We found that DD-9 treatment significantly reduced tumor cell proliferation in dose- and time-dependent manners in HepG2 and HeLa cells. The subsequent experiments in vitro implied that DD-63 could significantly suppress the tumor clonality, metastases, and induced apoptosis, and that it arrested the cell cycle at the G0/G1 phase of HepG2 and HeLa cells. Dual luciferase assay, Western blot, and immunofluorescence assay showed that DD-9 could dose-dependently attenuate the Wnt/β-catenin signaling by inhibiting β-catenin transcriptional activity and abrogating β-catenin translocated to the nucleus; down-regulating the transcription level of β-catenin-stimulated Wnt target gene and the expression of related proteins including p-GSK3-β, β-catenin, LEF1, Axin1, c-Myc, and CyclinD1; and up-regulating GSK3-β expression, which indicates that DD-9 stabilized the β-catenin degradation complex, thereby inducing β-catenin degradation and inactivation of the Wnt/β-catenin pathway. The possible interaction between DD-9 and β-catenin and GSK3-β protein was further confirmed by molecular docking studies. Collectively, DD-9 may suppress proliferation and induce apoptosis of liver and cervical cancer cells, possibly at least in part via GSK3-β-mediated crosstalk with the Wnt/β-catenin signaling axis, providing insights into the mechanism for the potency of DD-9 on hepatocellular and cervical cancer.

Anti-Toxic Principles from Musa Sapientum and Moringa Oleifera Ameliorated Skin Histo-Pathology, Decreased Lipid Peroxidation and Promoted Melatonin/TNF-Alpha/p53-Mediated Apoptosis in Cadmium Chloride-Induced Carcinogenesis in Rats

This study evaluated anticancer potentials of Musa sapientum (MS) and Moringa oleifera (MO) in cadmium chloride (CdCl2)-induced skin and liver carcinogenesis. Musa sapientum fraction-1 (MSF1) and Moringa oleifera fraction-6 (MOF6) were extracted from MS suckers and MO leaves, respectively, using column-chromatography methods. Forty-five adult male rats were randomly divided into 11 groups (n = 5). Cancer-induction was via single intraperitoneal administration of 1.25 mg/kg CdCl2. Groups 1 and 2 received physiological saline and CdCl2 dose, respectively. Groups 3–5 received CdCl2 dose on day 1 but were post-treated on days 15–56 with 15 mg/kg MOF6, 30 mg/kg MOF6 and 10 mg/kg MSF1, respectively. Group 6 received CdCl2dose on day 1 and were post-treated on days 15– 28 with doxorubicin + cisplatin doses. Groups 7–9 received MOF6 dose and MSF1 dose, respectively (days 1–56). Groups 10 and 11 received CdCl2dose on day 1 and were posttreated with 30 mg/kg MOF6 and 10 mg/kg MSF1, respectively (days 1–56). Doxorubicin and extracts doses were administered orally. Skin histo-pathology (haematoxylin and eosin), sera melatonin and tumour necrosis factor-alpha (TNFa) (enzyme linked immunosorbent assays [ELISA]) levels were evaluated. Malondialdehyde (thiobarbituric acid assay) and p53 (ELISA) levels were evaluated in liver homogenates. Data were statistically analysed using ANOVA at p ≤ 0.05. Results showed skin histo-alterations in Group 2, compared with normal skin histology in Groups 1 and 3–11. Statistical analyses showed significant downregulation of p53, non-significant downregulations of malondialdehyde and TNFa, and non-significant upregulation of melatonin in Groups 3–11 compared with Group 2. Overall, post-treatments with MOF6 and MSF1 exhibited possible anticancer potentials of these plants extracts via degrees of amelioration of CdCl2-induced skin histo-pathology and carcinogenesis.

Degradation kinetics of lycopene from red amaranth & preparation of winter melon jelly using this lycopene and comparison with commercial jelly

This study was conducted to observe the storage conditions, such as solvent and temperature, of lycopene content and degradation kinetics from red amaranth (Amaranthus gangeticus). Jelly was prepared using the extracted lycopene, the physicochemical properties and lycopene content. The extract with the maximum amount of lycopene was obtained by extraction with hexane, acetone and ethanol (2:1:1),50 ± 9 mg/kg. Higher lycopene degradation was observed at refrigerated temperature as compared to ambient temperature in hexane acetone (6:4) solvent throughout the storage periods. In this period, the initial lycopene concentration was measured to be 17 ± 8 mg/kg, whereas at the end of the storage time, it was found to be 3.0 ± 0.8 mg/kg. Hence, the results indicate that the hexane, acetone, and ethanol (in a ratio of 2:1:1) solvent method is viable for extracting and purifying lycopene from red amaranth at refrigerated temperature. This lycopene can serve as both a natural colorant and a value-added product. However, it is worth noting that lycopene can also be extracted and purified using recrystallization, column chromatography, and thin-layer chromatography (TLC) methods. The Winter melon jelly using lycopene from red amaranth contained moisture 29.6 %, ash 0.67 %, acidity 0.35 %, reducing sugar 26.8 %, non-reducing sugar 35.4 %, total soluble solid 66°brix and lycopene content 26.04 mg/kg. Proper utilization of lycopene extracted from red amaranth during the preparation of bakery, confectionary, baby food etc., may help and encourage the development of small-scale industries in the country.

Synthesis of silver nanoparticles from coffee parasite leaf extract (Scurrulla ferruginea (Roxb. ex Jack) Danser) and potential activity as an antioxidant and anticancer

This study aim is to know the results of the synthesis of silver nanoparticles from quercetin and Scurrula ferruginea (Roxb. Ex Jack) danser as well as its potential as an antioxidant and anticancer. The solvent used in the maceration extraction method in this research is the polar solvent methanol. Isolation and purification of thick extract compounds from coffee parasite leaves were carried out using liquid vacuum chromatography (CVC) and gravity column chromatography (GCC) methods. Silver (Ag) nanoparticle is made with the stirrer method at a speed of 1500 rpm for 60 minutes at a temperature of 70o C. Silver nanoparticle solution was produced and then characterized using FTIR (Fourier Transform Infra Red) and PSA (Particles Size Analyzer). PSA results analysis using extract coffee parasite size 214 nm particles, whereas using flavonoid isolate quercetin confirmed size 29 nm. FTIR showed that there was a stretching vibration of the OH group at a wave number of 3229.25 cm-1, the C=O function at the wave number 1637.87 cm-1, and the CO functional group at the wave number 1016.31 cm-1. Antioxidant test using the DPPH method was carried out in silver nanoparticle solution using extract Scurrula ferruginea (Roxb. Ex Jack) danser and with quercetin obtained an IC50 value of around 115.4 and 114.9 this result describes as moderate level of antioxidants. Even though the nanoparticle anticancer test was very weak, overall, the synthesis of silver nanoparticles using flavonoid isolates was better than using extracts from coffee parasite leaves.

High-Precision Measurements of 44Ca/40Ca Using Multiple-Collector Inductively Coupled Plasma Mass Spectrometry without Collision-Cell Technology.

Herein, we present a new method for determining the Ca isotopic composition of geological samples. To eliminate matrix elements from Ca, a column chromatography method was developed using a N,N,N'N' tetraoctyl-1,5-diglycolamide (TODGA) resin. The Ca isotopic compositions were measured by a multiple collector inductively coupled plasma mass spectrometry (MC-ICP-MS) without collision cell equipment, especially that direct measurement to 44Ca/40Ca can be achieved. To mitigate the interference from 40Ar during 40Ca measurement, the cold plasma technique was used to suppress the Ar+ generation, resulting in a background Ar+ intensity of <300 mV, in contrast to the conventional hot plasma conditions, which typically yield thousands of volts for Ar+ intensities. Given the potential for a concentration mismatch between the sample and bracketed standard solutions to cause an intensive shift in measured Ca isotopic compositions, a correction for the [Ca] match is needed. To avoid matrix effects arising from residue matrix elements, it is crucial to limit the concentrations below 1% of Ca for most matrix elements (including Al, Mg, K, Na, and Sr) and below 1‰ for Fe. Notably, the tolerance of residue Sr is effectively improved compared to measurements with CC-MC-ICP-MS and traditional Hot-plasma-SSB-MC-ICP-MS methods with the conventional hot plasma technique, thereby lowering the complexity of column chemistry. The measured δ44/40Ca, δ44/42Ca, and ε40Ca values for eight reference materials agree well with previously reported values within analytical uncertainties. This method demonstrates long-term precision is better than 0.10‰ (two standard deviations) for both δ values (i.e., δ44/40Ca and δ44/42Ca). We anticipate that the proposed method will benefit the growth of the Ca isotope data set and foster an increase in the application of Ca isotope in Earth science studies.

Antidiabetic Ligand Based Reverse Docking Studies on Isolated Phytoconstituents from Stem Bark Chloroform Extract of Strychnos nux‐vomica

AbstractStrychnos nux‐vomica (S.N) also known as poisonous nut, Dog button, Kuchla whose seeds and barks have various components that are used in folklore and traditional medicines in different countries. Stem bark of SN is collected from local area and extracted using chloroform by soxhlation process and isolated the phytoconstituents from the chloroform extract using column chromatography method by gradient elution technique. Six compounds are isolated from the column out of which four compounds are found to be novel from the S.N. Obtained phytoconstituents are checked for in silico studies using reverse docking process with the help of Schrodinger software and the docking score indicates that the phytoconstituents are found to have very good antidiabetic action.

Brick sand particles as an adsorbent in column chromatography to remove heavy metals from solution

The column chromatography method was used with brick sand particles as an adsorbent for the removal of heavy metals (Cr, Fe, and Pb) from the aqueous solution. These metals can pollute water, and soil and also come in contact with the food chain, causing life-threatening health issues in the human body. The pore volume and the specific surface area of the brick sand particles were determined using the standard method which was 44.41% and 29.4 m2/g respectively. The different concentrations of toxic metals in the water solution were considered to determine the adsorption capacity of the adsorbent. The adsorption methods were able to remove about 94% of Cr, 24% Pb, and 69% Fe from the aqueous solution. From the investigation results, we may conclude that raw brick sand particles can play a vital role as an adsorbent in the treatment of heavy metal-contaminated wastewater. Moreover, the saturated adsorbent can be used as an element of construction materials to prevent further pollution. Bangladesh J. Sci. Ind. Res. 59(1), 47-54, 2024

Plasma amino acids spectrum and Holter ECG monitoring indexes in coronary artery disease patients with atrial fibrillation

The research was conducted with the aim of establishing a connection between plasma amino acid peculiarities and Holter ECG monitoring indexes in Coronary Artery Disease (CAD) and Atrial Fibrillation (AF) patients. 300 patients were examined divided into 3 groups: I – 149 patients with CAD without arrhythmias, II – 124 patients with CAD and AF paroxysm, and the Control Group (CG) – 27 patients without CAD and arrhythmias. Holter ECG monitoring was performed for the patients with AF within 24 hours after sinus rhythm restoration, and for the patients without AF on the first day of observation. Plasma AA level was detected by the method of ion exchange liquid column chromatography. It was established that there is a significant increase glutamate and Branched-Chain Amino Acids (BCAA) levels in patients of the II group, and significant depletion in glycine levels comparison with the I group patients (p&lt;0.05). In the II group patients, an increase in SupraVentricular (SVE) and Ventricular Extrasystoles (VE) was checked in comparison with the I group patients (p&lt;0.05). Total SVE was significantly correlated with threonine (r=-0.316), serine (r=-0.336), glycine (r=-0.397), isoleucine (r=0.317), BCAA (r=0.356), and glycine+serine sum (r=-0.302), p&lt;0.05. AF paroxysm was significantly correlated with taurine (r=-0.302), serine (r=-0.328), glycine (r=-0.311), glutamine (r=-0.304), and glycine+serine sum (r=-0.379), p&lt;0.05. Total VE was significantly correlated with glycine (r=-0.370) and tyrosine (r=0.325), p&lt;0.05. Changes in ST-segment were significantly correlated with tyrosine (r=0.307), phenylalanine (r=0.318), and Aromatic Amino Acids (AAA) (r=0.379), p&lt;0.05. We concluded that glycine, serine, and BCAA are significantly correlated with cardiac arrhythmias. Changes in ST segment are significantly correlated with AAA levels. Keywords: heart rhythm violations, ischemia, arrhythmia, metabolomics.

Isolation and Identificationof Octadecane-3-on Compound From Ethyl Acetate Fraction of Momordica balsamina L. Leaves

Momordica balsamina L. is a wild plant that generally grows in moist soil and gets a lot of sunlight. The leaves and fruit are reported to have various medicinal and nutritional properties. This study aimed to isolate and identify the compounds contained in the ethyl acetate fraction in Momordica balsamina L The isolation stage was carried out by the maceration method and the separation and purification of the compounds using gravity column chromatography methods to obtain isolate fraction F1.6.2 with a sample weight of 2.4 mg. Identification of compounds using proton nuclear magnetic resonance (1H-NMR) and carbon nuclear magnetic resonance (13C-NMR) spectroscopy techniques. The proposed identified compound is octadecan-3-one or 3-octadecanone with the molecular formula C18H36O

Active compounds of Michelia champaca bark extract against Curvularia verruculosa fungi causing leaf spot disease in rice (Oryza sativa L.)

Curvularia verruculosa fungal causes leaf spot disease in rice plants. The bark extract of Michelia champaca could inhibit the growth of the fungi. The present research aimed to know the active compound responsible for antifungal activity. Extraction was done using the Maceration method, antifungal activity was measured using the Diffusion well method, and identification of active compounds was carried out using Gas chromatography-mass spectroscopy (GC-MS). The methanol extract obtained had a yield of 18.7%. It showed strong activity against C. verruculosa with an inhibition zone until 30.01 mm. The fractionation results showed that n-hexane extract (HE) was the strongest inhibiting the growth of C. verruculosa (32.45 mm), followed by chloroform extract (CE) (29.20 mm), while n-butanol extract (BE) was not active. Separating active compounds from HE extract was made using Column chromatography (CC) method with silica gel as the stationary phase and the mixture of n-hexane-acetone (3:1) as the mobile phase. This separation resulted in 5 combined fractions; HE3 and HE5 extracts showed very strong activity against C. verruculosa, with a diameter of the inhibition zone of 26.73 and 33.46 mm, respectively; HE2 extract showed strong activity with a diameter of the inhibition zone of 15.21 mm, while HE1 and HE4 extracts did not show activity. Identification using GC-MS, especially the HE3 extract, revealed that the extract contained two compounds: tributyl acetyl citrate and terephthalic acid, dodecyl-2-ethylhexyl ester. The result indicated that the bark extract of M. champaca had the potential to be a botanical fungicide.

A new bufadienolide from Bufo gargarizans Cantor

Bufo gargarizans Cantor (B. gargarizans) is the most widely distributed and abundant species of toad in China. Bufadienolides and indole alkaloids have cardiotonic and anti-tumor activities and are important pharmacological components of B. bufo gargarizans. In this experiment, a novel compound (1) and two known compounds (2 and 3) were isolated and identified from the dry skin of B. bufo gargarizans, both of which are bufadienolides. Various column chromatography methods were used to separate and purify the extract from the dried skin of B. bufo gargarizans. Accurate molecular weights were measured by HR-ESI-MS, and the chemical structure of the compounds was determined by NMR spectrometers. The structures were named as (2β,5β,16α)-2,5,16-trihydroxide bufa-14,20,22 dienolide (1), gamabufotalin (2) and desacetylbufotalin (3). In vitro cytotoxic activity assay indicated that compound 1 showed a moderate cytotoxicity against A549 cells with IC50 value of 12.65 μM.

ATRIAL FIBRILATION IN CORONARY ARTERY DISEASE PATIENTS: PLATELETS CHARACTERISTICS AND ECHOCARDIOGRAPHY INDEXES

The aim: To reveal the connections between platelets morphological and biochemical characteristics and echocardiography (TTE) indexes in patients with coronary artery disease (CAD) and atrial fibrillation (AF).&#x0D; Materials and methods: 300 patients were divided into 3 groups: first (CAD) – 149 patients with CAD but without arrhythmias, second (CAD+AF) – 124 patients with CAD and AF paroxysm and control group – 27 patients without CAD and arrhythmias. TTE was done by ALOKA SSD-5000. Platelets count (PC), mean platelet volume (MPV), platelets distribution width (PDW) and platelet-to-leucocyte ratio (PLR) was obtained by common blood count. Platelets amino acids spectrum was detected by method of ion exchange liquid column chromatography.&#x0D; Results: II group patients had significantly lower PC (12.20%) and higher MPV (9.30%) and PLR (41.12%) then in I group, P&lt;0.05. II group patients had significantly higher levels of platelets isoleucine (10.73%), leucine (12.63%) and lower threonine (23.05%), serine (5.06%), glycine (32.21%), valine (30.83%) levels then in I group, P&lt;0.05. II group patients had significantly higher levels of left atrium diameter (10.03%), left atrium volume (15.40%) and left atrium volume index (11.48%) then in I group, P &lt;0.05. Left atrium volume and left atrium volume index were corelated with PC (r=-0.315 and r=-0.444 consequently), MPV (r=0.327 and r=0.331 consequently), PLR (r=0.313 and r=0.303 consequently) and platelets serine (r=-0.302 and r=-0.310 consequently), alanine (r=-0.306 and r=-0.355 consequently) and leucine (r=0.346 and r=0.311 consequently), P&lt;0.05. Left ventricular mass and left ventricular mass index were corelated with MPV (r=0.308 and r=0.301 consequently), PLR (r=0.307 and r=0.305 consequently) and platelets arginine (r=0.366 and r=0.305 consequently), threonine (r=-0.474 and r=-0.383 consequently), serine (r=-0.387 and r=-0.361 consequently), alanine (r=-0.342 and r=-0.326 consequently), isoleucine (r=0.355 and r=0.328 consequently) and tyrosine (r=0.307 and r=0.321 consequently), P&lt;0.05.&#x0D; Conclusions: Left atrium size and left ventricular mass are connected with MPV, PLR and platelets amino acids content: serine, threonine, alanine, leucine, isoleucine and tyrosine levels.

A Simple, Low‐Blank Batch Purification Method for High‐Precision Boron Isotope Analysis

AbstractBoron (B) isotopes are widely used in the Earth sciences to trace processes ranging from slab recycling in the mantle to changes in ocean pH and atmospheric CO2. Boron isotope analysis is increasingly achieved by multi‐collector inductively coupled plasma mass spectrometry, which requires separation of B from the sample matrix. Traditional column chromatography methods for this separation have a well‐established track record but are time consuming and prone to contamination from airborne blank. Here, we present an extensive array of tests that establish a novel method for B purification using a batch method. We discuss the key controls and limitations on sample loading, matrix removal and B elution including sample volume, ionic strength, buffer to acid ratio and elution volume, all of which may also help optimize column‐based methods. We find consistent, low procedural blanks of 10 ± 16 pg and excellent reproducibility: 10 ng NIST RM 8301 foram [8301f] yields 14.58 ± 0.11‰ 2SD n = 15; 2.5 ng 8301f yields 14.60 ± 0.19‰ 2SD, n = 31; and overall long term 2SD on n = 218 samples pooling different sample sizes yields 14.62 ± 0.21‰ 2SD. This method also offers significant advantages in throughput, allowing the processing of 24 samples in ∼5 hr. This boron batch method thus provides a fast, reproducible, low‐blank method for purification of boron for high precision isotopic analyses.

Triterpene Compound from Ethyl Acetate Fraction of Kesambi Bark (Schleichera oleosa) and Its Activity as Anti-Bacterial

In our earlier study, we managed to find the antibacterial activity of the ethyl acetate fraction of the kesambi bark stem. In our next study, we isolated secondary metabolite compounds from the ethyl acetate fraction of the kesambi stem bark and tested the antibacterial activity of isolates against the bacteria Escherichia coli and Staphylococcus aureus. Separation is carried out by the method of column chromatography. Analysis of stain patterns and purity was performed with thin-layer chromatography (TLC). The test of antibacterial activity was carried out by the Kirby Bauer method. Characterization of pure isolates was performed with IR, 1H -NMR, 13C -NMR and DEPT 135° spectroscopy. The results of the characterization of pure isolates show that the isolated compound is a triterpenoid compound with the name IUPAC compound lup-20(29)-en-3-ol. The results of antibacterial tests showed that the compound lup-20(29)-en-3-ol has antibacterial activity against E. coli and S. aureus which is relatively moderate.

Optimization of culture conditions to express AA9 Polysaccharide monooxygenases AN3860 in Escherichia coli

Lignocellulose biomass is a copious source for second generation biomaterial production. The participant of Polysaccharide monooxygenases enzyme (PMO) in the reactions which convert lignocellulose biomass into monosaccharides enhances the activity and improve the efficiency of hydrolysis of hydrolase enzymes on lignocellulose substrate. Enzyme AN3860, obtained from Aspergillus nidulans strain belonging to AA9 PMO, is expected to catalyze flexibly at C1 and C4 carbon positions of β-glycosidic linkage. As an enzyme with high potential of improving cellulose crystals hydrolysis capacity, AN3860 was successfully cloned into the expression system of E. coli BL21 (DE3) strain. In this study, the culture process of recombinant strain with AN3680 gene is optimized to increase the target proteins yield, thus ensure the outcome of purification process, and save production cost. The results demonstrate that the E. coli recombinant strains grow sufficiently in TB (Terrific Broth) culture media and the highest yield of AN3680 protein achieved when the concentration of Isopropyl β-D-1-thiogalactopyranoside (IPTG) is 0.05 mM and the temperature of the reaction is 30 0C at 150 rpm. After 6 hours of induction, the biomass reaches 500 mg/L and the yield of AN3860 account for (7-10) % total protein generated. The recombinant AN3860 protein is later harvested on larger scale and purified by Ni-NTA column chromatography method for analysis of bioactivities on lignocellulose substrates in the future. ® 2022 Journal of Science and Technology - NTTU

ISOLASI DAN UJI AKTIVITAS ANTIBAKTERI SENYAWA METABOLIT SEKUNDER DARI EKSTRAK METANOL DAUN MARPUYAN (Rhodamnia cinerea Jack.)

A study on isolation and antibacterial activity of secondary metabolites from the methanol extract of Marpuyan (Rhodamnia cinerea Jack.) Leaves on Escherichia coli has been carried out. Isolation was carried out by column chromatography methods. Whereas the antibacterial activity test was carried out using agar diffusion method. The results of the isolation produced 17.9 mg of RC-F13 isolate compound in the form of light yellow crystals, with a melting point of 170-172oC and soluble in methanol. The results of identification of RC- F13 isolate compounds through testing with AlCl3 reagents, positive including the flavonoid group, were strengthened by the results of the UV spectrum and FT-IR spectrum. Based on the results of the UV spectrum, it was seen that the RC- F13 isolate compound showed λ max 353.60, 257.00 and 209.20 nm. The FT-IR spectrum shows the presence of several functional groups such as OH, C aliphatic C-H, C = O carbonyl, C = C aromatic and C-O. testing the antibacterial activity of methanol extract with a concentration of 20%; 10%; 1%; 0.1% and 0.01%, fraction 13 at 1% concentration; 0.1% and 0.01% and RC- F13 isolate compound concentration of 0.1%, against Escherichia coli bacteria did not show any antibacterial activity which was marked by the absence of clear areas around the disc.

Platelet amino acid spectrum and gut microbiota, their links in patients with coronary artery disease and atrial fibrillation

Background. The aim of our work was to identify the links between platelet amino acid (AA) spectrum and gut microbiota composition in patients with coronary artery disease (CAD) and atrial fibrillation (AF) and to evaluate them. Materials and methods. Three hundred patients were enrolled in the study. They were divided into 3 groups: I (CAD) — 149 patients with CAD but without arrhythmias, II (CAD + AF) — 124 people with CAD and AF paroxysm, and control group (CG) — 27 individuals without CAD and arrhythmias. Platelet AA level was assessed by method of ion-exchange liquid column chromatography. Gut microbiota composition was studied by 16-S rRNA sequencing. Results. A significant increase in isoleucine (10.73 %), leucine (12.63 %) and a decrease in threonine (23.05 %), serine (5.06 %), glycine (32.21 %) and valine (30.83 %) platelets levels was found in patients with CAD and AF (P &lt; 0.05). In addition, they had a significant increase in Bacteroides spp., Faecalibacterium prausnitzii, Actinobacter spp., Streptococcus spp., Ruminococcus spp. and a decrease in Lactobacillus spp., Bifidobacterium spp., Eubacterium rectale (P &lt; 0.05). Platelet glutamine acid, valine, glycine, asparagine acid, threonine had the highest number of significant correlations with gut microbiota species (P &lt; 0.05). Actinobacter spp., Blautia spp., Streptococcus spp., Akkermansia muciniphila and Roseburia inulinivorans had the highest number of significant correlations with platelet amino acids (P &lt; 0.05). Conclusions. Platelet amino acid spectrum and gut microbiota composition in patients with coronary artery disease and atrial fibrillation are closely linked.