Despite the proven therapeutic role of capsaicin in human health, its usage is still hampered by its high pungency. In this sense, nonpungent capsaicin analogues as olvanil are a feasible alternative ...to the unpleasant sensations produced by capsaicin while maintaining a similar pharmacological profile. Olvanil can be obtained by a lipase‐catalyzed chemoenzymatic process. In the present work, recombinant Candida antarctica lipase B (CALB) was expressed in Pichia pastoris and subsequently immobilized by cross‐linked enzyme aggregate (CLEA) methodology for the synthesis of olvanil. The CALB‐CLEAs were obtained directly from the fermentation broth of P. pastoris without any purification step in order to assess the role of the contaminant proteins of the crude extract as co‐feeders. The CALB‐CLEAs were also bioimprinted to enhance the catalytic performance in olvanil synthesis. When CALB was precipitated with isopropanol, the obtained CALB‐CLEAs exhibited the highest activity in the synthesis of olvanil, regardless of the glutaraldehyde concentration. The maximum product synthesis was found at 72 hr obtaining 6.8 g L−1 of olvanil with a reaction yield of 16%. When CALB was bioimprinted with olvanil, the synthesis was enhanced 1.3 times, reaching 10.7 g L−1 of olvanil at 72 hr of reaction with a reaction yield of 25%. Scanning electron microscopy images indicated different morphologies of the CLEAs depending on the precipitating agent and the template used for bioimprinting. Recombinant CALB‐CLEAs obtained directly from the fermentation broth are a suitable alternative to commercial enzymatic preparations for the synthesis of olvanil in organic medium.
In order to provide a second economic life to agave fibers, an important waste material from the production of tequila, filaments based on polylactic acid (PLA) were filled with agave fibers (0, 3, ...5, 10 wt%), and further utilized to produce biocomposites by fused deposition modeling (FDM)-based 3D printing at two raster angles (−45°/45° and 0°/90°). Differential scanning calorimetry, water uptake, density variation, morphology, and composting of the biocomposites were studied. The mechanical properties of the biocomposites (tensile, flexural, and Charpy impact properties) were determined following ASTM international norms. The addition of agave fibers to the filaments increased the crystallinity value from 23.7 to 44.1%. However, the fibers generated porous structures with a higher content of open cells and lower apparent densities than neat PLA pieces. The printing angle had a low significant effect on flexural and tensile properties, but directly affected the morphology of the printed biocomposites, positively influenced the impact strength, and slightly improved the absorption values for biocomposites printed at −45°/45°. Overall, increasing the concentrations of agave fibers had a detrimental effect on the mechanical properties of the biocomposites. The disintegration of the biocomposites under simulated composting conditions was slowed 1.6-fold with the addition of agave fibers, compared to neat PLA.
Background
Arylacetamide deacetylase (AADAC) is a lipolytic enzyme involved in xenobiotic metabolism. The characterization in terms of activity and substrate preference has been limited to a few ...mammalian species. The potential role and catalytic activities of AADAC from other organisms are still poorly understood. Therefore, in this work, the physicochemical properties, proteomic analysis, and protein-protein interactions from Gnathostomata organisms were investigated.
Results
The analysis were performed with 142 orthologue sequences with ~ 48–100% identity with human AADAC. The catalytic motif HGGA/G tetrapeptide block was conserved through all AADAC orthologues. Four variations were found in the consensus pentapeptide GXSXG sequence (GDSAG, GESAG, GDSSG, and GSSSG), and a novel motif YXLXP was found. The prediction of
N
-glycosylation sites projected 4, 1, 6, and 4 different patterns for amphibians, birds, mammals, and reptiles, respectively. The transmembrane regions of AADAC orthologues were not conserved among groups, and variations in the number and orientation of the active site and
C
-terminal carboxyl were observed among the sequences studied. The protein-protein interaction of AADAC orthologues were related to cancer, lipid, and xenobiotic metabolism genes.
Conclusion
The findings from this computational analysis offer new insight into one of the main enzymes involved in xenobiotic metabolism from mammals, reptiles, amphibians, and birds and its potential use in medical and veterinarian biotechnological approaches.
Graphical Abstract
In this work, cellulose nanocrystals (CNCs), bleached cellulose nanofibers (bCNFs), and unbleached cellulose nanofibers (ubCNFs) isolated by acid hydrolysis from Agave tequilana Weber var. Azul ...bagasse, an agro-waste from the tequila industry, were used as reinforcements in a thermoplastic starch matrix to obtain environmentally friendly materials that can substitute contaminant polymers. A robust characterization of starting materials and biocomposites was carried out. Biocomposite mechanical, thermal, and antibacterial properties were evaluated, as well as color, crystallinity, morphology, rugosity, lateral texture, electrical conductivity, chemical identity, solubility, and water vapor permeability. Pulp fibers and nanocelluloses were analyzed via SEM, TEM, and AFM. The water vapor permeability (WVP) decreased by up to 20.69% with the presence of CNCs. The solubility decreases with the presence of CNFs and CNCs. The addition of CNCs and CNFs increased the tensile strength and Young’s modulus and decreased the elongation at break. Biocomposites prepared with ubCNF showed the best tensile mechanical properties due to a better adhesion with the matrix. Images of bCNF-based biocomposites demonstrated that bCNFs are good reinforcing agents as the fibers were dispersed within the starch film and embedded within the matrix. Roughness increased with CNF content and decreased with CNC content. Films with CNCs did not show bacterial growth for Staphylococcus aureus and Escherichia coli. This study offers a new theoretical basis since it demonstrates that different proportions of bleached or unbleached nanofibers and nanocrystals can improve the properties of starch films.
A detailed study of the different structural transitions of the triblock copolymer PEO
-PPO
-PEO
(P104) in water, in the dilute and semi-dilute regions, is addressed here as a function of temperature ...and P104 concentration (C
) by mean of complimentary methods: viscosimetry, densimetry, dynamic light scattering, turbidimetry, polarized microscopy, and rheometry. The hydration profile was calculated through density and sound velocity measurements. It was possible to identify the regions where monomers exist, spherical micelle formation, elongated cylindrical micelles formation, clouding points, and liquid crystalline behavior. We report a partial phase diagram including information for P104 concentrations from 1 × 10
to 90 wt.% and temperatures from 20 to 75 °C that will be helpful for further interaction studies with hydrophobic molecules or active principles for drug delivery.
Capsaicin analogues are typically synthetized by condensation of the amide group of vanillylamine with a fatty acid derivative. The enzyme of choice to perform this reaction is
Candida antarctica
...lipase B; however, this enzyme is unable to react with the phenolic substiruents of the vanillyl ring. So far, this can only be achieved chemically, resulting in capsaicin esters with higher lipophilicity and less irritation than the parent compound. In need of searching for new capsaicin derivatives, we investigated the transesterification of the phenolic OH group of capsaicin and several capsaicin analogues by
Candida antarctica
lipase A (CALA) with vinyl esters. Capsaicin esters were successfully synthetized with a reaction yield of 80.6% and 57.5% with vinyl butyrate and vinyl laurate, respectively. When the reactions were performed with capsaicin analogues of different acyl chain lengths, CALA exhibited a noticeable transesterification preference for medium-length capsaicin analogues. In an attempt to explore the catalytic limits of CALA, we performed the transesterification of capsaicin from
Capsicum
oleoresin. The results showed similar transesterification yields to those obtained with semi-pure capsaicin. Within our knowledge, this is the first report to achieve the synthesis of phenolic OH esters of capsaicin and capsaicin analogues by enzymatic means.
•Olvanil inhibits adipogenesis of preadipocytes 3T3-L1.•Olvanil reduces lipid accumulation and triglycerides in 3T3-L1 adipocytes.•Olvanil decreases fat accumulation in the body of mice with ...DIO.•Olvanil improve lipid profile in serum of mice with DIO.
The present study is aimed to examine olvanil's effect in preadipocyte cell culture and on a murine model of diet-induced obesity. We hypothesized that olvanil by being a capsaicinoid will reduce the differentiation to mature adipocytes and reduce the weight of fat tissue in the studied mice.
To determine the effect of olvanil on adipogenesis, 3T3-L1 cells were cultured. Oil red staining was performed to determine lipid accumulation, whereas triglycerides were measured by biochemical determination. Expression of PPAR-γ and PREF-1 were measured by RT-PCR. Therefore, male C57BL/6J mice (CICUAL:2018-020B) were fed with a high-fat diet for 12 weeks to develop a murine model of diet-induced obesity (DIO). Animals were separated into 6 experimental groups: control (standard diet), DIO (high-fat diet), DIO + orlistat (10 mg/kg), DIO + olvanil (10 mg/kg), DIO + olvanil (25 mg/kg) and DIO + olvanil (50 mg/kg), olvanil was administered for 4 weeks.
Olvanil inhibits adipogenesis, reduces lipid accumulation and triglycerides in 3T3-L1 adipocytes. PPAR-ɣ gene expression was suppressed while PREF-1 was increased in adipocytes treated with olvanil. Whereas protein expression of FABP4 and PPAR-ɣ decreases significantly with olvanil. The results suggest that olvanil can inhibit the differentiation of preadipocytes to adipocytes through the overexpression or maintenance of PREF-1 levels and the suppression of PPAR-ɣ, and FABP4. Therefore, in diet-induced obesity in mice, olvanil decreases fat accumulation in the body and improve lipid profile by decreasing LDL, VLDL and triglycerides in serum.
Olvanil inhibits adipocyte differentiation in 3T3-L1 cells and reduces fat accumulation and ameliorate lipid profile in diet-induced obese mice.
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This study addresses the environmental problem of PET plastic through in silico bioprospecting for the identification and experimental validation of novel PET degrading eukaryotes through the in ...silico bioprospectingI of PETases, employing a methodology that combines Hidden Markov Models (HMMs), clustering techniques, molecular docking, and dynamic simulations. A total of 424 putative PETase sequences were identified from 219 eukaryotic organisms, highlighting six sequences with low affinity energies. The Aspergillus luchuensis sequence showed the lowest Gibbs free energy and exhibited stability at different temperatures in molecular dynamics assays. Experimental validation, through a plate clearance assay and HPLC, confirmed PETase activity in three wild-type fungal strains, with A. luchuensis showing the highest efficiency. The results obtained demonstrate the effectiveness of combining computational and experimental approaches as proof of concept to discover and validate eukaryotes with PET-degrading capabilities opening new perspectives for the sustainable management of this type of waste and contributing to its environmental mitigation.
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•In silico bioprospecting identified 219 eukaryotes with potential PETases.•Hidden Markov models retrieved 424 potential eukaryotic enzymes for PET degradation.•AluCut from A. luchuensis exhibits in silico thermostability and PETase activity.•Molecular dockings of AluCut with 2PET and 4PET showed desirable interactions.•These integrated methods confirmed PETase activity in targeted WT fungal strains.
A multilevel factorial design of 2
with 12 experiments was developed for the preparation of cellulose nanocrystals (CNC) from
Weber var. Azul bagasse, an agro-industrial waste from tequila ...production. The studied parameters were acid type (H
SO
and HCl), acid concentration (60 and 65 wt% for H
SO
, 2 and 8N for HCl) temperature (40 and 60 °C for H
SO
, 50 and 90 °C for HCl), and hydrolysis time (40, 55 and 70 min for H
SO
; and 30, 115 and 200 min for HCl). The obtained CNC were physical and chemically characterized using dynamic light scattering (DLS), atomic force microscopy (AFM), Fourier-transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and X-ray diffraction (XDR) techniques. The maximum CNC yield was 90 and 96% for HCL and H
SO
, respectively, and the crystallinity values ranged from 88-91%. The size and morphology of
CNC strongly depends on the acid type and hydrolysis time. The shortest CNC obtained with H
SO
(65 wt%, 40 °C, and 70 min) had a length of 137 ± 68 nm, width 33 ± 7 nm, and height 9.1 nm, whereas the shortest CNC obtained with HCl (2 N, 50 °C and 30 min) had a length of 216 ± 73 nm, width 69 ± 17 nm, and height 8.9 nm. In general, the obtained CNC had an ellipsoidal shape, whereas CNC prepared from H
SO
were shorter and thinner than those obtained with HCl. The total sulfate group content of CNC obtained with H
SO
increased with time, temperature, and acid concentration, exhibiting an exponential behavior of CSG=aebt.
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