Cholesterol Esterase Activity Research Articles

Inhibitory activity and molecular mechanism of the total alkaloids of Dendrobium officinale Kimura et Migo against cholesterol esterase

AbstractThis study aimed to explore the inhibitory effect and mechanism of the total alkaloids of Dendrobium officinale Kimura et Migo (DENA) against cholesterol esterase (CE). DENA was characterised by SEM, 1H NMR, and X-ray diffraction (XRD). The inhibitory effect and mechanism of DENA against CE were investigated through fluorescence chromatography, circular dichroism, and molecular docking. DENA inhibited CE activity (IC50 = 1.08 ± 0.09 mg mL−1), characterised by a non-competitive inhibition mechanism. Furthermore, DENA induced a fluorescence quenching in CE, causing a blue shift in the λmax. This coincided with a transition in the secondary structure of CE from a layered to a helical structure by circular dichroism, indicating a significant reduction in its stability. Moreover, molecular docking confirmed that DENA binds to amino acid residues in the enzyme through hydrogen bonds and hydrophobic interactions, leading to structural changes and reduced enzyme activity. These results suggest DENA has the potential to lower blood lipid levels by inhibiting CE activity.

Nanoliposome-Mediated Encapsulation of Chlorella Oil for the Development of a Controlled-Release Lipid-Lowering Formulation.

Chlorella oil nanoliposomes (CO-NLP) were synthesized through ultrasonic injection with ethanol, and their physicochemical properties and hypolipidemic efficacy were systematically investigated. The results revealed that the mean particle size of CO-NLP was 86.90 nm and the encapsulation efficiency (EE) was 92.84%. Storage conditions at 4 °C were conducive to the stability of CO-NLP, maintaining an EE of approximately 90% even after 10 days of storage. The release profile of CO-NLP adhered more closely to the first-order kinetic model during in vitro assessments, exhibiting a slower release rate compared to free microalgae oil. In simulated in vitro digestion experiments, lipolytic reactions of CO-NLP were observed during intestinal digestion subsequent to nanoliposome administration. Notably, the inhibitory effect of CO-NLP on cholesterol esterase activity was measured at 85.42%. Additionally, the average fluorescence intensity of nematodes in the CO-NLP group was 52.17% lower than in the control group at a CO-NLP concentration of 500 μg/mL, which suggests a pronounced lipid-lowering effect of CO-NLP. Therefore, the CO-NLP exhibited characteristics of small and uniform particle size, elevated storage stability, gradual release during intestinal digestion, and a noteworthy hypolipidemic effect. These findings designate CO-NLP as a novel lipid-lowering active product, demonstrating potential for the development of functional foods.

Ursolic Acid Inhibited Cholesterol Esterase and Pancreatic Lipase Activities and Decreased Micellar Cholesterol Solubility In Vitro

Ursolic acid (UA) is a natural triterpene carboxylic acid with an underlying anti-hyperlipidemia effect. This study examined the mechanism of interactions of UA with cholesterol esterase (CEase), pancreatic lipase (PL), and micellar cholesterol solubility in vitro. The half-maximal inhibitory concentration (IC50) of CEase, PL, and micellar cholesterol solubility was determined to be 0.07 ± 0.01 mg/mL, 1.81 ± 0.13 mg/mL, and 1.73 ± 0.08 mg/mL, respectively. Multispectral combination revealed that UA changed the secondary structure and quenched the intrinsic fluorescence by static quenching of the two enzymes. The interactions were exothermic reaction as determined by enthalpy. Furthermore, molecular docking confirmed that UA was bound to amino acids of two enzymes at active site through hydrophobic interaction and van der Waals forces. Molecular dynamics (MD) simulation found that CEase and PL rearranged with UA to form stable complexes. The strong inhibition effect of CEase and PL mediated by UA might provide additional insight into understanding the role of UA in the hypolipidemic effect.

Exploring the inhibitory potential of KPHs-AL-derived GLLF peptide on pancreatic lipase and cholesterol esterase activities

The effective modulation of pancreatic lipase and cholesterol esterase activities proves critical in maintaining circulatory triglycerides and cholesterol levels within physiological boundaries. In this study, peptides derived from KPHs-AL, produced through the enzymatic hydrolysis of skipjack tuna dark muscle using alkaline protease, have a specific inhibitory effect on pancreatic lipase and cholesterol esterase. It is hypothesized that these peptides target and modulate the activities of enzymes by inducing conformational changes within their binding pockets, potentially impacting the catalytic functions of both pancreatic lipase and cholesterol esterase. Results revealed these peptides including AINDPFIDL, FLGM, GLLF and WGPL, were found to nestle into the binding site groove of pancreatic lipase and cholesterol esterase. Among these, GLLF stood out, demonstrating potent inhibition with IC50 values of 0.1891 mg/mL and 0.2534 mg/mL for pancreatic lipase and cholesterol esterase, respectively. The kinetics studies suggested that GLLF competed effectively with substrates for the enzyme active sites. Spectroscopic analyses, including ultraviolet–visible, fluorescence quenching, and circular dichroism, indicated that GLLF binding induced conformational changes within the enzymes, likely through hydrogen bond formation and hydrophobic interactions, thereby increasing structural flexibility. Molecular docking and molecular dynamics simulations supported these findings, showing GLLF's stable interaction with vital active site residues. These findings position GLLF as a potent inhibitor of key digestive enzymes, offering insights into its role in regulating lipid metabolism and highlighting its potential as functional ingredient.

Triterpenoids from Protorhus longifolia Exhibit Hypocholesterolemic Potential via Regulation of Cholesterol Biosynthesis and Stimulation of Low-Density Lipoprotein Uptake in HepG2 Cells.

The increasing incidence of hypercholesterolemia-related diseases even in the presence of the currently available cholesterol-lowering drugs indicates a need to discover new therapeutic drugs. This study aimed to investigate the hypocholesterolemic potential of two triterpenoids isolated from Protorhus longifolia stem bark. In silico techniques and in vitro enzyme assays were used to evaluate the potential inhibition of cholesterol esterase and HMG-CoA reductase by the triterpenoids (ARM-2 and RA-5). The toxicity, modulation of low-density lipoprotein (LDL) uptake, and associated gene expression were determined in HepG2 hepatocytes. In silico molecular docking revealed that ARM-2 compared with RA-5 has a relatively stronger binding affinity for both enzymes. Both triterpenoids further demonstrated promising in silico drug-likeness properties and favorable ADMET profiles characterized by high intestinal absorption and lack of CYP450 enzyme inhibition. The compounds further showed, to varying degrees of efficacy, inhibition of cholesterol micellization as well as both cholesterol esterase and HMG-CoA reductase activities with IC50 values ranging from 16.4 to 41.1 μM. Moreover, enhanced hepatic cellular LDL uptake and the associated upregulation of the LDL-R and SREBP-2 gene expression were observed in the triterpenoid-treated HepG2 cells. It is evident that the triterpenoids, especially ARM-2, possess hypocholesterolemic properties, and these molecules can serve as leads or structural templates for the development of new hypocholesterolemic drugs.

Inhibitory effect of procyanidin B2 and tannin acid on cholesterol esterase and their synergistic effect with orlistat

This study was aimed to analyze the effect of procyanidin B2 (PC) and tannin acid (TA) on the activities of cholesterol esterase (CEase) and the inhibitory mechanisms of enzymatic activity. The interaction mechanisms were investigated by enzymatic kinetics, multi-spectroscopy methods, thermodynamics analysis, molecular docking, and dynamic simulations. PC and TA could bind with CEase and inhibit the activity of enzyme in a mixed-competitive manner and non-competitive manner, which was verified by molecular docking simulations and dynamics simulations. Also, PC and TA showed the synergistic inhibition with orlistat. Fluorescence, UV-vis and the thermodynamic analysis revealed that the complexes were formed from CEase and inhibitors by noncovalent interaction. As revealed by the circular dichroism results, both PC and TA decreased enzymatic activities by altering the conformations of CEase. The inhibition of PC and TA on CEase might be one mechanism for its cholesterol-lowering effect.

Chemical characterization of Passiflora edulis extracts and their in vitro antioxidant, anti-inflammatory, anti-lipid activities, and ex-vivo vasodilation effect

Passion fruit is a dietary plant with the protective proterties against many diseases such as cardiovascular diseases. The aim of this study was to explore the biological properties (anti-oxidant, anti-inflammatory, anti-lipid, and vascular relaxation) of the P. edulis extracts obtained from seed (PSEE) and fruit (PFWE). Thin-layer chromatography (TLC) and HPLC were used to analyze the phytochemical constituents in the extracts. Antioxidant and enzyme inhibition properties were examined. Anti-inflammatory activities of PFWE and PSEE were evaluated in lipopolysaccharide (LPS)-induced macrophages and ex-vivo vascular relaxation were also determined. A high contents of piceatannol and polyphenolic stilbene were found in PSEE extracts. On the other hand, beta-carotene and gamma-tocopherol were predominant in PFWE. PFWE exhibited stronger anti-oxidant activity than PSEE. Both PSEE and PFWE showed significant inhibition of lipase and cholesterol esterase activities. However, only PFWE showed minimal inhibitory effect against HMG-CoA reductase. PSEE and PFWE caused a significant reduction of nitric oxide level in LPS-induced inflammation in RAW264.7 cells. Both extracts also showed ex-vivo vasorelaxation of rat aortic rings. It has been demonstrated that PFWE and PSEE possesses anti-oxidant activity, anti-inflammatory activity, pancreatic lipase and cholesterol esterase, and ex-vivo vasorelaxation of rat aortic rings, however, the activity of HMG-CoA reductase was inhibited by only PFWE. These findings suggest the beneficial effect of PSEE and PFWE against hyperlipidemia thus preventing the development of hypertension. However, further investigations in animal are needed to elucidate.

Assessment of the Hypoglycemic and Hypolipidemic Activity of Flavonoid-Rich Extract from Angelica keiskei.

Angelica keiskei contains a variety of bioactive compounds including chalcone, coumarin, and phytochemicals, endowing it with pharmacological effects such as lipid-lowering activity, antitumor activity, liver protection, and nerve protection. This study aims to study the hypoglycemic and hypolipidemic effects of the flavonoid-rich extract from Angelica keiskei (FEAK) in an effort to exploit new applications of FEAK and increase its commercial value. In this paper, flavonoid compounds in Angelica keiskei were extracted using 50% ethanol, and the contents of the flavonoid compounds were analyzed by UPLC-MS/MS. Then, the hypoglycemic and hypolipidemic activities of the FEAK were investigated through in vitro enzyme activity and cell experiments as well as establishing in vivo zebrafish and Caenorhabditis elegans (C. elegans) models. The UPLC-MS/MS results show that the major flavonoid compounds in the FEAK were aureusidin, xanthoangelol, kaempferol, luteolin, and quercetin. The inhibitory rates of the FEAK on the activity of α-amylase and cholesterol esterase were 57.13% and 72.11%, respectively. In cell lipid-lowering experiments, the FEAK significantly reduced the total cholesterol (TC) and total triglyceride (TG) levels in a dose-dependent manner, with 150 μg/mL of FEAK decreasing the intracellular levels of TC and TG by 33.86% and 27.89%, respectively. The fluorescence intensity of the FEAK group was 68.12% higher than that of the control group, indicating that the FEAK exhibited hypoglycemic effects. When the concentration of the FEAK reached 500 μg/mL, the hypoglycemic effect on zebrafish reached up to 57.7%, and the average fluorescence intensity of C. elegans in the FEAK group was 17% lower than that of the control group. The results indicate that the FEAK had hypoglycemic and hypolipidemic activities. The findings of this study provide theoretical references for the high-value utilization of Angelica keiskei and the development of natural functional food with hypoglycemic and hypolipidemic activities.

Insight into structural characteristics of theabrownin from Pingwu Fuzhuan brick tea and its hypolipidemic activity based on the in vivo zebrafish and in vitro lipid digestion and absorption models

Fuzhuan brick tea (FBT) is a post-fermented dark tea preferred by consumers for its excellent hypolipidemic activity, and theabrownin (TB) is the main bioactive composition in FBT. This work explored the structural and hypolipidemic properties of TB derived from Pingwu FBT, and investigated whether it exerted hypolipidemic activity by inhibiting intestinal lipid absorption. Structural characterization revealed that TB was an amorphous polymerized phenolic compound rich in hydroxyl and carboxyl groups with good thermostability. In vivo, TB and its fractions with different molecular weights (TB-LT3k, TB-3-10k, TB-10-30k, TB-30-100k, TB-GT100k) significantly reduced the lipid levels of hyperlipidemia zebrafish (P < 0.05). Moreover, TB attenuated hyperlipidemia by inhibiting intestinal lipid absorption, as TB effectively bound to bile acids, inhibited enzymatic activity of pancreatic lipase and cholesterol esterase, influenced micelle formation, and decreased micellar cholesterol solubility. Results facilitated research on TB and offered support for its feasibility as a natural alternative to prevent hyperlipidemia.

Cholesterol esterase substantially enhances phytosterol ester bioaccessibility in a modified INFOGEST digestion model

Standardization and of in vitro digestion models is crucial to obtain results that may be compared across laboratories. The INFOGEST network has published multiple iterations of a static in vitro digestion protocol. The protocol lacks cholesterol esterase (CE) activity during the intestinal phase, and few studies have evaluated its incorporation into the model. In the present study, the effect of additional CE on phytosterol ester (PSE) bioaccessibility in emulsions with different droplet sizes was evaluated. Different droplet sizes were obtained by utilizing different homogenization techniques to produce large (9.9 μm), medium (3.4 μm), and fine (1.3 μm) emulsions. Addition of CE resulted in an increase in PSE bioaccessibility for all emulsions. This change was emphasized in emulsions with smaller droplet sizes, with bioaccessibility of large (19.6%), medium (34.2%), and fine (40.8%) emulsions increasing 1.5, 2.2, and 2.1-fold. This finding provides evidence for the inclusion of CE into the INFOGEST protocol.

In Vitro Antioxidant, Anti-Inflammatory, and Digestive Enzymes Inhibition Activities of Hydro-Ethanolic Leaf and Bark Extracts of Psychotria densinervia (K. Krause) Verdc.

Psychotria densinervia hydro-ethanolic leaf extract (PHELE) and bark extract (PHEBE) were evaluated for antioxidant, anti-inflammatory, and inhibition of digestive enzymes activities. The antioxidant activity was characterized by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), total phenolic content (TPC), and total flavonoid content (TFC) assays. The anti-inflammatory activity was characterized by protein denaturation and antiproteinase tests, while the inhibition of the enzymes was assessed using α-amylase, α-glucosidase, lipase, and cholesterol esterase activities. PHELE presented low (p < 0.001) IC50 (59.09 ± 5.97 μg/ml) for DPPH compared with ascorbic acid (71.78 ± 6.37 μg/ml) and PHEBE (115.40 ± 1.21 μg/ml). The IC50 of PHELE (262.4 ± 4.46 μg/ml) and PHEBE (354.2 ± 1.97 μg/ml) was higher (p < 0.001) than that of catechin (33.48 ± 2.02 μg/ml) for ABTS. PHELE had high (p < 0.001) FRAP (341.73 ± 21.70 mg CE/g) than PHEBE (150.30 ± 0.32 mg CE/g). PHELE presented (p < 0.001) high TPC (270.05 ± 7.53 mg CE/g) and TFC (23.43 ± 0.032 mg CE/g) than PHEBE (TPC: 138.89 ± 0.91 and TFC: 20.06 ± 0.032 mg CE/g). PHELE showed antiprotein denaturation with IC50 (257.0 ± 7.51 μg/ml) (p < 0.001) and antiproteinase activity (74.37 ± 1.10 μg/ml) lower than PHEBE (316.1 ± 6.02 μg/ml and 177.6 ± 0.50 μg/ml), respectively. Orlistat inhibited lipase (p < 0.001) activity with IC50 (37.11 ± 4.39 μg/ml) lower than PHELE and PHEBE (50.57 ± 2.89 μg/ml and 62.88 ± 1.74 μg/ml, respectively). PHELE inhibited cholesterol esterase with IC50 (34.75 ± 3.87 μg/ml) lower than orlistat (54.61 ± 2.56) and PHEBE (80.14 ± 1.71 μg/ml). PHELE inhibited α-amylase IC50 (6.07 ± 4.05 μg/ml) lower than PHEBE (19.69 ± 6.27 μg/ml) and acarbose (20.01 ± 2.84 μg/ml). Acarbose inhibited α-glucosidase (p < 0.001) activity with IC50 (4.11 ± 3.47 μg/ml) lower than PHELE (24.41 ± 2.84 μg/ml) and PHEBE (38.81 ± 2.46 μg/ml). PHELE presented better antioxidant, anti-inflammatory, and enzyme inhibition activity than PHEBE.

Evaluation of in vitro Cholesterol Esterase Inhibitory Activity of Few Selected Vegetables – An Indication of Cardioprotective Property

Pharmacognosy Research,2022,14,2,188-194.DOI:10.5530/pres.14.2.27Published:April 2022Type:Original ArticleAuthors:Thiyagarajan Sathishkumar, Muruvanda Bhuvan Uthaiah, Ravichandran Vinu Varshith, Kuppamuthu Kumaresan, and Vinohar Stephen Rapheal Author(s) affiliations:Thiyagarajan Sathishkumar, Muruvanda Bhuvan Uthaiah*, Ravichandran Vinu Varshith, Kuppamuthu Kumaresan, Vinohar Stephen Rapheal Department of Biotechnology, Kumaraguru College of Technology, Coimbatore, Tamil Nadu, INDIA. Abstract:Background: Cardiovascular diseases are responsible for severe mortality rate amongst the population. Several drugs available to control and/ or cure the same, cause severe adverse effects amidst human population. Contextually, the present study was devised to screen the in vitro cholesterol esterase (CEase) activity of few selected vegetables as an indicator of cardioprotective property. Objectives: To screen and purify the polyphenolic fractionate from the vegetable(s) extract that record an optimal CEase activity. Materials and Methods: CEase inhibitory activity was investigated using standard spectrophotometric assay. Microwave assisted central composite design based response surface methodology (CCD based RSM) was executed to quantify an improvement in the CEase inhibitory activity of Solanum melongena extract. One dimensional thin layer chromatography (1D TLC) and two-dimensional preparative thin layer chromatography (2D PTLC) analyses were carried out for Brassica oleracea var. botrytis hot water extract to identify and eluate the polyphenolic fractionate, respectively. Results: Significant CEase inhibitory activity was noticed in the hot water extract of Brassica oleracea var. botrytis (84.65 ± 3.29%) than the other vegetable extracts. Even though the optimal process model was validated (at 5% level), no significant improvement was recorded in the CEase inhibition. The presence of polyphenolic content in the hot water extract of Brassica oleracea var. botrytis (Rf = 0.69) was confirmed by comparing with standard quercetin (Rf = 0.60). Strong blue and moderate brownish yellow color spots revealed the presence of phenolic acid and flavonoid eluates respectively, were purified using 2D PTLC technique. The eluate has recorded a near moderate (28.57 ± 2.12%) CEase inhibitory activity. Conclusion: Brassica oleracea var. botrytis possess significant CEase inhibitory activity and proved to be a good diet to protect from various cardiovascular diseases. Key words: Brassica oleracea var. botrytis, Cholesterol esterase, Solanum melongena, Microwave Assisted Response Surface Methodology, Flavonoid. View:PDF (245.05 KB)

Piper nigrum and Morinda lucida Possess Antioxidant Capacities and Regulate the Activities of Key Carbohydrate and Lipid Digestive Enzymes

Introduction: Carbohydrate and lipid digestive enzymes are instrumental in the absorbability of nutrients associated to diabetes and obesity. This study evaluated hydroethanolic extracts of Piper nigrum leaf and Morinda lucida stem bark for antioxidant capacity and enzymes (carbohydrate and lipid digestive) inhibition.&#x0D; Methods: Colorimetric assays determined enzyme (α-amylase, α-glucosidase, lipase and cholesterol esterase) inhibition and antioxidant capacity (total phenolic (TPC) and flavonoid (TFC) content, radical scavenging activity (DPPH, ABTS), and ferric reducing antioxidant power (FRAP)) of hydroethanolic ethanolic extracts, ethyl acetate and hexane fractions.&#x0D; Results: At 1 mg/ml extracts of P nigrum and M lucida inhibited α- amylase (9.82±1.05 - 36.63±0.69 %) and α-glucosidase (22.47±0.34 - 67.77±0.58 %) activities. At 100 µg/ml extracts and fractions inhibited lipase (56.72±1.11 - 81.61±0.71 %) and cholesterol esterase (18.14 ±0.79 - 36.84±0.70 %) activities. IC50 for α- amylase (2.20±0.02 - 7.8±1.42 mg/ml), α-glucosidase (0.16±0.01 - 3.74±0.01 mg/ml), lipase (8.58±2.57 - 53.03±5.20 µg/ml) and cholesterol esterase (172.20±5.12 - 419.80±4.55 µg/ml) were registered. At 4 mg/ml, P. nigrum presented a higher TPC (153.78±8.31 - 354.63±6.33 mg/ml), TFC (21.65±1.14 -33.86±0.00 mg/ml) than M lucida TPC (10.21±0.11 - 169.89±6.54 mg/ml), TFC (ND - 87.32±6.14 mg/ml). P nigrum presented radical scavenging (DPPH and ABTS) activity with IC50 0.12±0.00 - 1.27±0.01 mg/ml compared to 1.31±0.02 - 3.44±0.12 mg/ml of M lucida. The FRAP IC50 values were better for P nigrum (3.38±0.14- 4.48±1.05 mg/ml) than M lucida (3.34±1.32 - 15.4±2.03 mg/ml).&#x0D; Conclusion: P nigrum presented better antioxidant capacity and more effective on lipid digestive enzymes while M lucida was more effective on carbohydrate digestive enzymes.

Effect of Abutilon indicum (L) Extract on Adipogenesis, Lipolysis and Cholesterol Esterase in 3T3-L1 Adipocyte Cell Lines.

Abutilon indicum (L) is an Indian traditional plant used for the treatment of diabetes and heart diseases. The present study is to evaluate the functional of A. indicum leaf extract as insulin like character to inhibit lipolysis and stimulates Adipogenesis activity. The ability of the A. indicum leaf extract in anti-obesity effect of Adipogenesis, lipolysis and cholesterol esterase functions can be predicted by using 3T3-L1 adipocyte cell lines. Substances were isolated from A. indicum leaves and the double filtered crude sample were used for Adipogenesis, lipolysis and cholesterol esterase activity using 3T3-L1 adipocytes at different concentrations. We used differential media-I, differential media-II and maintenance media (MM1) at concentrations of 20, 40, 60, 80, 100, 200 and 400µg/mL respectively. In addition to the extract, there is a significance increase in glycerol release (p < 0.001) compared with crude and reference compounds. Cholesterol esterase activity predicts the IC50 = 27.11µg/mL of orlistat positive control compare with IC50 = 8.158µg/mL of crude extract. Based on the observation, A. indicum leaf extract can promotes lipolysis and differentiated adipocytes. It is potentially used as adjuvant in the treatment of Type 2 diabetes.

Identification and characterization of cholesterol esterase and lipase inhibitory peptides from amaranth protein hydrolysates

Human diet is undergoing a shift towards plant-based diet as a sustainable source of protein compared to animal-derived protein. In this study, cholesterol esterase (CEase) and pancreatic lipase (PL) inhibitory activities of amaranth protein hydrolysates (APHs) were studied. Bromelain, chymotrypsin, and actinase E were used for generating APHs at 2, 4 & 6 h of hydrolysis. Higher PL inhibiting potential were observed in bromelain-derived APHs (IC50 = 0.38–0.66 mg/mL) in comparison to intact amaranth proteins (IC50 = 3.93 mg/mL). Bromelain-4 h hydrolysates (AB4) demonstrated significant inhibitory potential for both CEase (IC50 = 0.47 mg/mL) and PL (IC50 = 0.48 mg/mL) activity. Peptide identification in AB-4 hydrolysate revealed that among 17 bioactive peptides, three peptides (FPFPPTLGY, FGAPR, and FPFVPAPT) were predicted as potential PL inhibitors and only one peptide (FPFVPAPT) was predicted as CEase inhibitor based on the number of substrate binding sites on active site of the enzymes. This is the first study providing insights into amaranth protein derived bioactive peptide possessing CEase and LIP inhibitory potential.

Inhibitory in vitro effects of Basil ( Ocimum basilicum ) leaf extracts on cholesterol esterase activity and the growth of Escherichia coli

The present study was to analyze the in vitro antibacterial activity and the effects on cholesterol esterase activity of the methanol extract from basil leaves and its hexane, ethyl acetate, butanol, and aqueous fractions. The highest total phenolic and flavonoid contents were recorded in the methanol extract. Total 92 compounds were determined by using gas chromatography-mass spectrometry belong to hydrocarbons, fatty acids, alcohols, esters, and phenols which reportedly have a variety of biological activities such as antibacterial, antiviral, anti-inflammatory, and anti-carcinogenic activities. This study evaluated the antibacterial activity of the basil methanol extract and its fractions based on the inhibitory concentrations of Escherichia coli at 50 mg/ml. The best results were obtained with the methanol extract, hexane, and ethyl acetate fractions, which exhibited anti-cholesterol esterase activity. Novelty impact statement The highest phenolic content, flavonoid content, and cholesterol esterase inhibitory activity were detected in the methanol extract. Ninety-two compounds were identified using gas chromatography-mass spectrometry from Thai Basi. Methanol extract, hexane, and ethyl acetate fractions had strong antibacterial activity at 50 mg/ml.

An In Vitro Study on the Effects of Selected Natural Dietary Fiber from Salad Vegetables for Lowering Intestinal Glucose and Lipid Absorption.

Salad vegetables are good sources of dietary fiber and are becoming increasingly popular among consumers. Therefore, these plants have the potential to be developed as functional foods. Using an in vitro model, this study investigated the physical properties and intestinal glucose and lipid absorption capacities of dry dietary fiber from vegetables typically consumed in salads (types of lettuce, including red oak, red coral, green oak, butterhead, and cos). Fiber was prepared from each type of lettuce using an enzymatic method and then characterized. Physical properties, including solubility and water-binding, swelling, cation-exchange, and oil-binding capacities, and antihyperglycemic and antihypercholesterolemic effects of fiber were investigated. The hydration capacity of total dietary fiber and insoluble fiber from the majority of sources was significantly different from that of cellulose. Adsorption and diffusion of glucose were directly proportional to incubation time, and the diffusion rate was significantly lower in the treatments containing fiber compared to the cellulose control. Fiber from these vegetables also inhibited amylase and alpha-glucosidase activities. Moreover, fiber from all sources exhibited significantly higher sodium cholate and cholesterol-binding capacity compared to cellulose, and also retarded pancreatic cholesterol esterase activity in a concentration-dependent manner. This study demonstrates that natural dietary fiber from salad vegetables can reduce glucose and lipid absorption and breakdown rates, thus preventing increases in postprandial blood glucose and cholesterol levels, which can be beneficial to human health.

Phenolic Profile of Nipa Palm Vinegar and Evaluation of Its Antilipidemic Activities.

Obesity and overweight are strongly associated with dyslipidemia which can promote the development of cardiovascular diseases. Recently, natural products have been suggested as alternative compounds for antioxidant and antilipidemic activity. The objective of this study was to determine the phenolic compounds and assess the inhibitory activities on pancreatic lipase, cholesterol esterase, and cholesterol micellization of nipa palm vinegar (NPV). Total phenolic content was assessed and phenolic compounds were determined using the Folin–Ciocalteu assay and liquid chromatography-mass spectrometry (LC-MS), respectively. Pancreatic lipase and cholesterol esterase inhibitory activities of the NPV were measured using enzymatic colorimetric assays. The formation of cholesterol micelles was assessed using a cholesterol assay kit. The phenolic content of NPV was 167.10 ± 10.15 µg GAE/mL, and LC-MS analyses indicated the presence of gallic acid, isoquercetin, quercetin, catechin, and rutin as bioactive compounds. Additionally, the NPV inhibited pancreatic lipase and cholesterol esterase activities in a concentration-dependent manner. Moreover, the NPV also suppressed the formation of cholesterol micellization. These results suggest that phenolic compounds, especially gallic acid, isoquercetin, quercetin, catechin, and rutin, from NPV may be the main active compounds with possible cholesterol-lowering effects through inhibition of pancreatic lipase and cholesterol esterase activities as well as the inhibition of solubility of cholesterol micelles. Therefore, NPV may delay postprandial dyslipidemia, and it could be used as a natural source of bioactive compounds with antilipidemic activity. However, NPV should be extensively evaluated by animal and clinical human studies.

Isolation and identification of cholesterol esterase and pancreatic lipase inhibitory peptides from brewer's spent grain by consecutive chromatography and mass spectrometry.

The isolation and identification of cholesterol esterase (CE) and pancreatic lipase (PL) inhibitory peptides obtained from the protein hydrolysate of brewer's spent grain (BSG) was performed. BSG peptides were fractionated and purified sequentially by anion exchange, gel filtration (FPLC), and reversed phase high-performance liquid chromatography (RP-HPLC). The fractions obtained from each chromatographic step were collected and the in vitro enzyme inhibitory activity was evaluated. The chromatographic purification process increased the in vitro activities. The most active fractions were evaluated using MALDI-TOF tandem mass spectrometry, which identified three peptides: a peptide with the highest CE inhibition capacity (WNIHMEHQDLTTME) and two peptides with PL inhibition capacity (DFGIASF and LAAVEALSTNG). These three peptides showed hydrophobic and acidic amino acid residues (Asp and Glu) and/or their amines (Asn and Gln), which could be a common feature among lipid-lowering peptides related to CE and PL enzyme inhibition. The in silico studies showed that the three peptides had high hydrophobicity and were susceptible to enzymatic hydrolysis performed by trypsin, pepsin, and pancreatin. The BSG byproduct was a good source of CE and PL inhibitory peptides, thus adding value to this byproduct of the beer industry. This is the first report to demonstrate that BSG peptides can inhibit CE and PL enzymes.

Production of recombinant extracellular cholesterol esterase using consistently active promoters in Burkholderia stabilis.

Burkholderia stabilis FERMP-21014 produces highly active cholesterol esterase in the presence of fatty acids. To develop an overexpression system for cholesterol esterase production, we carried out RNA sequencing analyses to screen strongly active promoters in FERMP-21014. Based on gene expression consistency analysis, we selected nine genes that were consistently expressed at high levels, following which we constructed expression vectors using their promoter sequences and achieved overproduction of extracellular cholesterol esterase under fatty acid-free conditions. Of the tested promoters, the promoter of BSFP_0720, which encodes the alkyl hydroperoxide reductase subunit AhpC, resulted in the highest cholesterol esterase activity (24.3 U mL-1). This activity level was 243-fold higher than that of the wild-type strain under fatty acid-free conditions. We confirmed that cholesterol esterase was secreted without excessive accumulation within the cells. The gene expression consistency analysis will be useful to screen promoters applicable to the overexpression of other industrially important enzymes.