Binding Of Lipase Research Articles

Structural insights into polyethylene terephthalate (PET)-degrading archaeal lipase enzyme: An insilico approach

Biodegradation of plastic is the most eco-friendly degradation process conferred by various microbial enzymes, especially the lipase enzymes. The lipase enzymes depict a broad substrate range, high regio/stereoselectivity, and stability in organic solvents. However, enhancing the catalytic activity of lipases for industrial production remains a challenge, predominantly due to the need for their extensive mechanistic characterization. Here, we employed a multidisciplinary approach to illustrate the comprehensive physiochemical and mechanical properties of 102 lipase sequences from six different groups of plastic-degrading microorganisms. Molecular docking-based evaluation of the binding efficiencies of selected lipases with the most common plastic polymers- polyurethane (PUR) and polyethylene terephthalate (PET), revealed that the archaeal lipase had the lowest binding energy of −7.1 kcal/mol with PET. Remarkably, the molecular dynamic simulation studies (up to 100 ns) and very low RMSD (0.3 nm) and RMSF (0.25 nm) values further ascertained the stability of archaeal lipase after binding with PET polymer. Further, a 2.19 nm radius of gyration (Rg) within 100 ns simulation time, binding free energy ΔGTOTAL with −24.36 kcal/mol and entropy factor -TΔS 7.65 kcal/mol also indicated strong binding of archaeal lipase with PET polymer. The mechanistic exploration of archaeal lipase yielded valuable insights into its effective binding with PET polymer, paving the way for future lipase enzyme engineering and harnessing its industrial potential for plastic biodegradation.

Controlling release of astaxanthin in β-sitosterol oleogel-based emulsions via different self-assembled mechanisms and composition of the oleogelators

In this study, three types of β-sitosterol-based oleogels (β-sitosterol + γ-oryzanol oleogels, β-sitosterol + lecithin, oleogels and β-sitosterol + monostearate oleogels), loaded with astaxanthin, were employed as the oil phase to create oleogel-based emulsions (SO, SL, and SM) using high-pressure homogenization. The microstructure revealed that fine-scale crystals were dispersed within the oil phase of the droplets in the β-sitosterol oleogel-based emulsion. The bioaccessibility of astaxanthin was found to be 58.13 %, 51.24 %, 36.57 %, and 45.72 % for SM, SL, SO, and the control group, respectively. Interestingly, the release of fatty acids was positively correlated with the availability of astaxanthin (P = 0.981). Further analysis of FFAs release and kinetics indicated that the structural strength of the oil-phase in the emulsions influenced the degree and rate of lipolysis. Additionally, the micellar fraction analysis suggested that the nature and composition of the oleogelators in SM and SL also impacted lipolysis and the bioaccessibility of astaxanthin. Furthermore, interfacial binding of lipase and isothermal titration calorimetry (ITC) measurements revealed that the oleogel network within the oil phase of the emulsion acted as a physical barrier, hindering the interaction between lipase and lipid. Overall, β-sitosterol oleogel-based emulsions offer a versatile platform for delivering hydrophobic molecules, enhancing the bioavailability of active compounds, and achieving sustained release.

Design, Synthesis, Characterization, Enzyme Inhibition, Molecular Docking, and Pharmacological Evaluation of New Chalcone‐Sulfonate Derivatives Bearing Thiophene

AbstractThe novel chalcone‐sulfonate derivatives bearing thiophene motif were synthesized and characterized using 1H NMR, 13C NMR, and HRMS analysis. The evaluation of in vitro and in silico potential pancreatic lipase inhibition activity of the novel chalcone‐sulfonate derivatives bearing thiophene motif was scanned. IC50 values of compounds 5 i (28.76±2.11 μM) and 5 f (30.58±0.45 μM) were determined to be more effective pancreatic lipase inhibitors for in vitro studies. The best potential inhibitor for pancreatic lipase binding affinity was found as compound 5 f (−9.8 kcal mol−1) for in silico studies. Although compounds 5 f and 5 i were identified as the best pancreatic lipase inhibitor candidates in vitro and molecular docking studies, compounds 5 f and 5 i were predicted mutagenic and carcinogenic properties in mice according to ADMET studies. Deeply, compound 5 h was a more effective pancreatic lipase inhibitor according to enzyme inhibition, molecular docking, and ADMET studies. It can be said that compound 5 h may be a more efficient drug candidate than orlistat in the treatment of obesity.

Mechanism for improving the in vitro digestive properties of coconut milk by modifying the structure and properties of coconut proteins with monosodium glutamate

In this paper, the effect of monosodium glutamate (MSG) on coconut protein (CP) solubility, surface hydrophobicity, emulsification activity, ultraviolet spectroscopy and fluorescence spectroscopy was investigated. Meanwhile, the changes in the in vitro digestive properties of coconut milk were also further analyzed. MSG treatment altered the solubility and surface hydrophobicity of CP, thereby improving protein digestibility. Molecular docking showed that CP bound to pepsin and trypsin mainly through hydrogen bonds and salt bridges. And MSG increased the cleavable sites of pepsin and trypsin on CP, thus further improving the protein digestibility. In addition, MSG increased the Na+ concentration in coconut milk, promoted flocculation and aggregation between coconut milk droplets, which prevented the binding of lipase and oil droplets and inhibited lipid digestion. These findings may provide new ideas and insights to improve the digestive properties of plant-based milk.

Activated carbon from sugarcane bagasse as support for lipase immobilization by physical adsorption technique

Lipases are recognized as the most important group of catalysts in biotechnology. However, utilization of free enzymes is often hampered by the need for more operational stability, high cost, and non-reusability. Most of these obstacles can be solved by lipase immobilization. This work's objective was to evaluate the performance of the activated carbon obtained from sugarcane straw (SAC) as a support for lipase immobilization. Two lipases were immobilized by physical adsorption on SAC: Aspergillus niger 11T53A14 lipase and CalB (lipase B from Candida antarctica, Novozymes). Results revealed that the lipase had been anchored on the activated carbon with the lipase binding efficiency of 89 % (A. niger lipase) and 100 % (CalB) at the optimum experimental conditions (initial protein concentration 0.1 mg mL-1, 0.15 g of SAC, 25 °C, and 120 min). Langmuir isotherm fitted the adsorption equilibrium data of the lipases on SAC. SAC presents a high surface area and protein adsorption capacity. These results show that activated carbon synthesized from the sugarcane straw is a promising support for enzyme immobilization.

A silver cluster-assembled material as a matrix for enzyme immobilization towards a highly efficient biocatalyst.

Silver cluster-assembled materials (SCAMs) epitomize well-defined extended crystalline frameworks that combine the ingenious designability at the atomic/molecular level and high structural robustness. They have captivated the interest of the scientific fraternity because of their modular construction which enables to systematically tailor their functions, and their capacity to not only inherit the characteristics of component building units but also introduce their uniqueness in endowing the final material with extraordinary properties. Herein, we demonstrate the synthesis of a novel (3,6)-connected two-dimensional (2D) SCAM [Ag12(StBu)6(CF3COO)6(THIT)6]n (described as TUS 5, THIT = 2,4,6-tri(1H-imidazol-1-yl)-1,3,5-triazine) composed of Ag12 cluster nodes and tritopic imidazolyl linkers. We have leveraged, for the first time, this precisely architected extended SCAM structure as a support matrix for enzyme immobilization. The electrostatic attraction between the negatively charged amano lipase PS and positively charged TUS 5 as well as the surface hydrophobicity of TUS 5 catered to great binding of lipase onto the TUS 5 matrix, in addition to boosting the activity of lipase via interfacial activation. Capitalizing on the cooperative benefits of organic and inorganic support matrices wherein organic supports impart with cost-efficiency, biocompatibility, and improved enzyme stability and reusability and inorganic supports confer high thermal, mechanical and microbial resistance, we have utilized the immobilized lipase on TUS 5 SCAM (lipase@TUS 5) for the kinetic resolution of (R,S)-1-phenylethanol by transesterification reaction. Importantly, lipase@TUS 5 could attain appreciably higher conversion into (R)-1-phenylethyl acetate, besides featuring superior thermal stability, solvent tolerance and recyclability, over the native lipase.

Production of high quality biodiesel from sunflower soapstock acid oil as novel feedstock: Catalyzed by immobilized pancreatic lipase

AbstractEdible refined oils, which are utilized as raw materials in biodiesel production, have been replaced by by‐products (acid oil, fatty acid, deodorized distillate, and soapstock distillates) obtained from the vegetable oil refining industry, in recent years. This study aims to investigate the production of high quality biodiesel fuel in accordance with the standards (TS EN and ASTM) from sunflower soapstock acid oil. This feedstock was donated by an oil factory in the Thrace region, Turkey. Esterification reaction was performed by immobilized enzyme. For this, immobilization was carried out by covalent binding of pancreatic lipase in glutaraldehyde activated chitosan and optimum immobilization conditions were determined. The activity of the immobilized lipase and the retained activity were found to be 65.69 U/μg and 61.8%, respectively (pH: 7.5, 37°C). The Km (Michealis constant) and Vmax (maximum velocity of an enzymatically catalyzed reaction) values of the immobilized enzyme were found to be 5.1 mmol/L and 48.6 U/min/mg protein, respectively. The immobilized enzyme was employed as a biocatalyst for esterification of sunflower soapstock acid oil. Notably, an impressive yield of 75.6% was attained under the conditions of a 1:5 molar ratio of soapstock acid oil to methanol, with 10 wt% immobilized lipase as the catalyst, and a reaction temperature of 45°C for 36 h. The resulting biodiesel exhibits fuel characteristics that meet the standards outlined in TS EN 14214:2012 + A2 and ASTM D6751‐02.

Maximizing Efficiency and Affordability with Ceramic Membranes for Enzyme Immobilization

AbstractLow‐cost ceramic microfiltration membranes were explored for immobilizing lipases. Expensive commercial ceramics (e.g., alumina, zirconia) have advantages like chemical robustness, but high sintering temperatures limit their usability. To overcome this, researchers used Fuller's earth clay with rice husk ash to create cost‐effective membranes. However, fewer reactive sites on ceramic surfaces hinder enzyme immobilization. To address this, 3‐aminopropyltriethoxysilane and glutaraldehyde activated the membrane for covalent lipase binding. The immobilized membrane retained activity for over five 1‐h reaction cycles with a minimal performance decline of around 60 %. After five cycles, it retained over 40 % of the initial activity. These results show promise for using low‐cost ceramic membranes in various industries, such as in food, beverages, pharmaceuticals, and chemical production.

Effect of wheat aleurone on lard emulsions during in vitro digestion

Dietary wheat aleurone has been shown to affect lipid metabolism and reduce the incidence of obesity. However, the underlying mechanisms are not fully understood. This work aimed to investigate how whole wheat aleurone affects lipolysis during the whole digestion process in vitro. The physicochemical and microstructural changes and the lipolysis kinetics of different lard emulsion mixtures were determined. The results showed that the lipolysis rate and degree are inversely proportional to the amount of wheat aleurone. Wheat aleurone and flour promoted the aggregation and flocculation of lipid droplets by increasing the viscosity. More importantly, the dietary fibers released from aleurone digestion can reduced the binding of lipase to lipid droplets by adsorbing lipid droplets to increase the steric hindrance effect. These results provide a better understanding of how whole grains affect lipid digestibility and will further contribute to the development of functional foods and the improvement of individual health.

Immobilization of Candida Rugosa Lipase on the Glutaraldehyde-Activated Chitosan Beads

An immobilized enzyme is a biocatalyst that speeds up the conversion of a chemical reaction. The application of enzymes for chemical synthesis is an effort toward a responsible production initiative to ensure the sustainability of chemical synthesis. Therefore, in the present work, Candida rugosa lipase was immobilized onto glutaraldehyde-activated chitosan beads through covalent bond linkages. Chitosan is biodegradable and contains amine groups, which serve as bases for lipase binding via cross-linking with bifunctional cross-linkers like glutaraldehyde. The immobilization of lipase on the chitosan beads was confirmed by determining lipase activity through the hydrolysis of a standard substrate. The effect of lipase and glutaraldehyde concentrations on the immobilization and activity yield was investigated. In general, lipase and glutaraldehyde concentrations have a significant effect on immobilization and activity yield. The interaction between the investigated parameters is significant toward the activity yield rather than the immobilization yield. The optimized immobilization procedures give lipase activity up to 46 IU by using 0.013 g/mL lipase and 2 %/v/v glutaraldehyde. It was found that the immobilized enzyme was rather stable and could be recycled 7 times. Therefore, immobilization of lipase onto glutaraldehyde-activated chitosan support is feasible.

Interaction of fungal lipase with potential phytotherapeutics.

Interaction of thymol, carvacrol and linalool with fungal lipase and Human Serum Albumin (HSA) have been investigated employing UV-Vis spectroscopy Fluorescence and Circular dichroism spectroscopy (CD) along with docking studies. Thymol, carvacrol and linalool displayed approximately 50% inhibition at 1.5 mmol/litre concentrations using para-nitrophenyl palmitate (pNPP). UV-Vis spectroscopy give evidence of the formation of lipase-linalool, lipase-carvacrol and lipase—thymol complex at the ground state. Three molecules also showed complex formation with HSA at the ground state. Fluorescence spectroscopy shows strong binding of lipase to thymol (Ka of 2.6 x 109 M-1) as compared to carvacrol (4.66 x 107 M-1) and linalool (5.3 x 103 M-1). Number of binding sites showing stoichiometry of association process on lipase is found to be 2.52 (thymol) compared to 2.04 (carvacrol) and 1.12 (linalool). Secondary structure analysis by CD spectroscopy results, following 24 hours incubation at 25°C, with thymol, carvacrol and linalool revealed decrease in negative ellipticity for lipase indicating loss in helical structure as compared with the native protein. The lowering in negative ellipticity was in the order of thymol > carvacrol > linalool. Fluorescence spectra following binding of all three molecules with HSA caused blue shift which suggests the compaction of the HSA structure. Association constant of thymol and HSA is 9.6 x 108 M-1 which along with ‘n’ value of 2.41 suggests strong association and stable complex formation, association constant for carvacrol and linalool was in range of 107 and 103 respectively. Docking results give further insight into strong binding of thymol, carvacrol and linalool with lipase having free energy of binding as -7.1 kcal/mol, -5.0 kcal/mol and -5.2 kcal/mol respectively. To conclude, fungal lipases can be attractive target for controlling their growth and pathogenicity. Employing UV-Vis, Fluorescence and Circular dichroism spectroscopy we have shown that thymol, carvacrol and linalool strongly bind and disrupt structure of fungal lipase, these three phytochemicals also bind well with HSA. Based on disruption of lipase structure and its binding nature with HSA, we concluded thymol as a best anti-lipase molecule among three molecules tested. Results of Fluorescence and CD spectroscopy taken together suggests that thymol and carvacrol are profound disrupter of lipase structure.

Taguchi design-assisted co-immobilization of lipase A and B from Candida antarctica onto chitosan: Characterization, kinetic resolution application, and docking studies

In the present communication, the simultaneous co-immobilization by covalent binding of lipase A from Candida antarctica (CALA) and lipase B from Candida antarctica (CALB) in glutaraldehyde (GLU) activated chitosan (CHI) was optimized using the Taguchi method. Under optimized conditions (pH 9, 5 mM, 6:1 (protein load/g of support and 1 h), it was possible to reach 80.00 ± 0.01% for the immobilization yield (IY) and 46.01 ± 0.35 U/g for the activity of the derivative (AtD); in this case, load protein and ionic strength were the only statistically significant parameters and, therefore, those that most influenced the immobilization process. Furthermore, at pH 7, CALA-CALB-CHI had a half-life 2–6 times longer than the mixture of CALA and CALB for a temperature range of 50−80 °C. CALA-CALB showed the highest activity at pH 7, whereas CALA-CALB-CHI, except at pH 7, was more active than the soluble lipase mixture in the pH range (5–9), especially at pH 9. CHI, CHI-GLU, and CALA-CALB-CHI were characterized by X-ray powder diffraction (XRPD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscope (SEM), Thermogravimetry (TGA), and Energy Dispersive Spectroscopy (EDS), proving the immobilization of CALA and CALB in chitosan. CALA-CALB-CHI derivative evaluated in the kinetic resolution of halohydrins acetates rac-2-bromo-1-(2-chlorophenyl) ethyl acetate (2a) and rac-2-chloro-1-(2,4-dichlorophenyl) ethyl acetate (2b), to produce the corresponding halohydrins 3a-b, which are intermediates in the synthesis of the drugs chlorprelanine (antiarrhythmic) and luliconazol (antifungal), respectively. (S)-bromohydrin 3a was obtained with 79% enantiomeric excess (ee), whereas (S)-chlorohydrin 3b produced with 98% ee, conversion of 46% and E > 200. Additionally, molecular docking was performed to elucidate the hydrolysis interaction reaction between β-halohydrin acetates and lipases CALA-CALB.

Bionic-immobilized recombinant lipase obtained via bio-silicification and its catalytic performance in biodiesel production

In this study, bio-silicification was employed to develop a green and simple bionic-immobilization method for the immobilization of recombinant lipase. The results showed that under the optimized bionic-immobilization conditions, a maximum immobilization efficiency and an activity recovery of 97.71% and 89.62%, respectively, were obtained. Further, bionic-immobilization significantly improved the thermal stability, storage stability, and short-chain alcohol resistance of the bionic-immobilized lipase. Furthermore, molecular dynamics simulation of the bionic-immobilization mechanism revealed that the key amino acid residues that facilitated the binding of the recombinant lipase to silica particles included Arg336, Lys345, His391, His392, His394, and His395, indicating that hydrogen bonds had a special effect in intermolecular recognition, and the active center of the bionic-immobilized lipase favored substrate entry due to the conformational changes of Phe291 and Ile320. Finally, biodiesel was prepared using the bionic-immobilized lipase as catalyst, and after optimizing the enzymatic process, the biodiesel yield reached 91.04% using 20% immobilized lipase dosage within 12 h at 45 °C, and a catalytic activity of 55.02% was observed following the reutilization of the bionic-immobilized lipase seven times. These findings suggest that bionic-immobilization is a novel and rapid method to realize recombinant enzyme immobilization, and can provide economic and ecological support for the biocatalytic preparation of industrial products.

Fabrication of immobilized Candida rugosa lipase on magnetic Fe3O4-poly(glycidyl methacrylate-co-methacrylic acid) composite as an efficient and recyclable biocatalyst for enzymatic production of biodiesel

Abstract The development of a magnetically recyclable immobilized lipase for enzymatic biodiesel production is of great research value to meet the need of green and sustainable development. In this work, the organic copolymer, poly(glycidyl methacrylate-co-methacrylic acid) (poly(GMA-co-MAA)), was encapsulated onto the Fe3O4 nanoparticles, and then Candida rugosa lipase was bound on the magnetic composites through coupling reactions with epoxy and carboxyl groups. The Fe3O4-poly(GMA-co-MAA) composite and immobilized lipase were characterized by several techniques, including enzyme activity assay, FT-IR,XRD,SEM,TEM,XPS,VSM and nitrogen adsorption–desorption techniques. Results revealed that the Fe3O4-poly(GMA-co-MAA) composites were successfully synthesized, and the lipase had been anchored on the magnetic composites with the lipase binding efficiency of 88.7% and activity recovery of 67.3%. The immobilized lipase displayed well-defined structure and good magnetic responsibility with saturation magnetization of 28.73 emu/g, and furthermore showed high catalytic activities to the transesterification of soybean oil. The biodiesel yield of 92.8% was attained through three-step addition of methanol at 40 °C with this magnetic biocatalyst. The immobilized lipase is recovered facilely by magnetic separation, still retaining at 79.4% of biodiesel yield after reuse for five cycles, and thus highlighting its great potential in the sustainable production of biodiesel.

The influence of equine body weight gain on inflammatory cytokine expressions of adipose tissue in response to endotoxin challenge

BackgroundHuman obesity is linked with systemic inflammation. However, it is still controversial if equines produce more inflammatory cytokines with increasing body weight and if the production of those show breed type specific patterns. The main objective of this study was to determine if diet induced obesity is associated with increased inflammatory signatures in adipose tissue of equines and if a breed predisposition exists between ponies and horses. Additionally, we aimed to identify adipose tissue depot differences in inflammatory cytokine expression. Nineteen healthy, non-overweight and metabolically healthy equines received a hypercaloric diet for 2 years. Body weight, body condition score and cresty neck score were assessed weekly throughout the study. At three time points, insulin sensitivity was determined by a combined glucose-insulin test. Adipose tissue samples were collected from two intra-abdominal and two subcutaneous depots under general anesthesia at each time point after an endotoxin trigger. In the adipose tissue samples levels of CD68 mRNA (a marker of macrophage infiltration) and pro-inflammatory cytokine mRNA (IL-1β, IL-6 and TNFα) were analyzed with RT-qPCR. As markers of lipid metabolism mRNA levels of lipoprotein lipase (LPL) and fatty acid binding protein 4 (FABP4) were determined with RT-qPCR.ResultsCD68 mRNA levels increased with body weight gain in several adipose tissue (AT) depots (Wilcoxon signed rank test with Bonferroni correction; retroperitoneal AT horses: P = 0.023, mesocolonial AT horses: P = 0.023, subcutaneous tail head AT ponies: P = 0.015). In both abdominal depots CD68 mRNA levels were higher than in subcutaneous adipose tissue depots (Kruskal–Wallis-ANOVA with Bonferroni correction: P < 0.05). No breed related differences were found. Pro-inflammatory cytokine mRNA IL-1β, IL-6 and TNFα levels were higher in subcutaneous depots compared to abdominal depots after body weight gain. IL-1β, IL-6 and TNFα mRNA levels of mesocolon adipose tissue were higher in obese horses compared to obese ponies (Mann–Whitney-U test; IL-1β: P = 0.006; IL-6: P = 0.003; TNFα: P = 0.049). In general, horses had higher FABP4 and LPL mRNA levels compared to ponies in neck AT and tail AT at all time points.ConclusionOur findings suggest an increased invasion of macrophages in intra-abdominal adipose tissue with increasing body weight gain in equines in combination with a low dose endotoxin stimulus. This might predispose equines to obesity related comorbidities. In obese horses mesocolon adipose tissue showed higher inflammatory cytokine expression compared to obese ponies. Additionally, subcutaneous adipose tissue expressed more pro-inflammatory cytokines compared to intra-abdominal adipose tissue. Horses had higher FABP4 and LPL mRNA levels in selected AT depots which may indicate a higher fat storage capacity than in ponies. The differences in lipid storage might be associated with a higher susceptibility to obesity-related comorbidities in ponies in comparison to horses.

Unfolding of monomeric lipoprotein lipase by ANGPTL4: Insight into the regulation of plasma triglyceride metabolism

The binding of lipoprotein lipase (LPL) to GPIHBP1 focuses the intravascular hydrolysis of triglyceride-rich lipoproteins on the surface of capillary endothelial cells. This process provides essential lipid nutrients for vital tissues (e.g., heart, skeletal muscle, and adipose tissue). Deficiencies in either LPL or GPIHBP1 impair triglyceride hydrolysis, resulting in severe hypertriglyceridemia. The activity of LPL in tissues is regulated by angiopoietin-like proteins 3, 4, and 8 (ANGPTL). Dogma has held that these ANGPTLs inactivate LPL by converting LPL homodimers into monomers, rendering them highly susceptible to spontaneous unfolding and loss of enzymatic activity. Here, we show that binding of an LPL-specific monoclonal antibody (5D2) to the tryptophan-rich lipid-binding loop in the carboxyl terminus of LPL prevents homodimer formation and forces LPL into a monomeric state. Of note, 5D2-bound LPL monomers are as stable as LPL homodimers (i.e., they are not more prone to unfolding), but they remain highly susceptible to ANGPTL4-catalyzed unfolding and inactivation. Binding of GPIHBP1 to LPL alone or to 5D2-bound LPL counteracts ANGPTL4-mediated unfolding of LPL. In conclusion, ANGPTL4-mediated inactivation of LPL, accomplished by catalyzing the unfolding of LPL, does not require the conversion of LPL homodimers into monomers. Thus, our findings necessitate changes to long-standing dogma on mechanisms for LPL inactivation by ANGPTL proteins. At the same time, our findings align well with insights into LPL function from the recent crystal structure of the LPL•GPIHBP1 complex.

Lipase immobilized on magnetic hierarchically porous carbon materials as a versatile tool for the synthesis of bioactive quercetin derivatives

The preparation, characterization and application of a novel robust nanobiocatalyst, developed through the covalent binding of lipase B from Pseudozyma antarctica on magnetic hierarchically porous carbon materials (HPCFe), is reported. The nanobiocatalyst was characterized by combination of spectroscopic and microscopic techniques. Structural and catalytic characterization indicates that HPCFe nanostructures create a microenvironment which stabilizes the structure of the immobilized enzyme, resulting in enhanced activity and stability in non-aqueous media over other forms of the biocatalyst. The nanobiocatalyst was effectively applied for the selective deacetylation of peracetylated quercetin towards the synthesis of 3,5,7-triacetoxy-3′,4′-dihydroxyflavone, a compound with high antiproliferative activity. At the optimum bioprocess conditions, the produced amount of the bioactive quercetin derivative in a single-step process reached values up to 3.48 g L−1 which is 13 times higher than that reported to date. The immobilized enzyme retains ~100% of its catalytic activity after 10 repeated reaction cycles.

Bio-ceramic, mesoporous cuttlebone of Sepia officinalis is an ideal support for the immobilization of Bacillus subtilis AKL13 lipase: optimization, adsorption, thermodynamic and reaction kinetic studies

Mesoporous cuttlebone powder of Sepia officinalis (CBP) was characterized and used as matrix for the immobilization of Bacillus subtilis AKL 13 lipase (BsL). Particle size and surface area of the matrix used for enzyme immobilization were 89.95 µm and 1.631 m2 g−1, respectively. Remarkable thermostability (54% of weight loss at 700 °C) of CBP was determined in TGA profile. The lipase immobilization process parameters were sequentially optimized by response surface methodology followed by artificial neural network and genetic algorithm. The maximum lipase loading capacity of CBP was 255 mg g−1 of support. Immobilized lipase (CBP-BsL) showed maximum specific activity of 5808 U mg−1 of protein in p-nitrophenol palmitate hydrolysis. Adsorption isotherm study revealed that the binding of lipase on the surface of CBP was Langmuir and the binding was physical adsorption. CBP-BsL showed lower activation energy (51.4 KJ mol−1) and higher thermal stability with half-lives of 13.3 h at 50 °C. Higher activity retention in nonpolar solvents and 69% of operational stability after 15 cycle of reaction were measured.

Betatrophin/Lipasin/C19orf80: In Silico Approach for Protein-Based Biomaterial Marker in Metabolic Syndrome and Colorectal Cancer Through Computational-Based Study

This computational material-based study aimed to provide a comprehensive data with the molecular interaction between betatrophin and its target as the confirmation of the primary role of betatrophin on serological lipid profile alteration and metabolic syndrome progression. We analyzed the binding behavior of betatrophin and LPL using docking model. The underlying molecular interaction between betatrophin and lipoprotein lipase (LPL) binding site results in the inhibitory activity of betatrophin on LPL. Our in Silico data showed that betatrophin could regulate that essential enzyme and suggested indirectly to restrict TG level during the circadian cycle in the human circulation. To sum up, betatrophin or RIFL shows as a promising future early biomarker for obesity and cancer linked metabolic syndrome. Hence, this preliminary computational material analysis data provided a hallmark for the future development of betatrophin as the clinical biomaterial marker in metabolic syndrome and gastrointestinal cancer.

Basic study on lipase-immobilized magnetic nanoparticles

Herein, the magnetic nanoparticles (MNPs) were produced by the solgel technique. This magnetic carrier was characterized by transmission electron microscopy, Fourier-transform infrared spectroscopy and X-ray diffraction. The size of the MNPs was determined to be 10.33 nm from the diffraction patterns, and also, least agglomeration was observed in TEM. The presence of required peaks in FT-IR confirmed MNPs formation, and their significant shifts confirmed the lipase binding. Then, lipase was immobilized onto MNPs by covalent attachment following surface modification and activation. The study reported maximum immobilization percentage as 92.3% which reduced on successive lipase loadings. Excellent thermostability and reusability were exhibited by the lipase-bound MNPs in the activity assay. At optimum pH (7.5), relative activity of the immobilized lipase showed no notable change, whereas, at higher temperature, it was higher than that of free lipase. This can be inferred that good thermal resistance was provided by the MNPs to lipase from unconditional disturbances. Increased Vmax showed the negligible steric hindrance in the active sites of immobilized lipase. At the eighth cycle of the process, it was observed that enzyme attained half of its maximum activity. Hence, this magnetite carrier may be considered as the best choice for immobilization of industrially important enzyme such as lipase and their efficient application.