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1.
Food Res Int ; 174(Pt 1): 113521, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37986510

RESUMO

The differences in wheat flour characteristics caused by ancient (pestle and mortar), old (stone hand mill), and modern (roller and cyclone) milling techniques and their effect on in vitro starch digestibility of wheat porridge using the simulated TIM Gastrointestinal Model (TIM-1) were investigated. Ancient flour (AF) was the coarsest flour (∼70 % is >1000 µm), followed by old wholemeal flour (OWF) and old refined flour (ORF) with similar particle size distribution showing one prominent peak (at ∼1000 µm for OWF and ∼800 µm for ORF). Modern refined flour (MRF) had a monomodal distribution centered at a particle size of ∼100 µm, while modern wholemeal flour (MWF) particle size was distributed between 40 and 600 µm. MRF and MWF porridges had higher cumulative sugar bioaccessibility than OWF and AF porridges, with ORF porridge having an intermediate cumulative sugar bioaccessibility. Characterizing the cumulative sugar bioaccessibility profile with a shifted logistic model allows identifying that the maximum sugar bioaccessibility and rate of sugar release were significantly higher (p < 0.05) for MRF and MWF compared to OWF and AF porridges, while the induction times were shorter, demonstrating the importance of processing on modulating starch digestibility.


Assuntos
Açúcares , Triticum , Farinha , Digestão , Amido
2.
Gels ; 9(8)2023 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-37623103

RESUMO

Bigels have been mainly applied in the pharmaceutical sector for the controlled release of drugs or therapeutics. However, these systems, with their intricate structures, hold great promise for wider application in food products. Besides their classical role as carrier and target delivery vehicles for molecules of interest, bigels may also be valuable tools for building complex food structures. In the context of reducing or even eliminating undesirable (but often highly functional) food components, current strategies often critically affect food structure and palatability. The production of solid fat systems that are trans-fat-free and have high levels of unsaturated fatty acids is one of the challenges the food industry currently faces. According to recent studies, bigels can be successfully used as ingredients for total or partial solid fat replacement in complex food matrices. This review aims to critically assess current research on bigels in food and pharmaceutical applications, discuss the role of bigel composition and production parameters on the characteristics of bigels and further expand the use of bigels as solid fat replacers and functional food ingredients. The hydrogel:oleogel ratio, selected gelators, inclusion of surfactants and encapsulation of molecules of interest, and process parameters (e.g., temperature, shear rate) during bigel production play a crucial role in the bigel's rheological and textural properties, microstructure, release characteristics, biocompatibility, and stability. Besides exploring the role of these parameters in bigel production, future research directions for bigels in a food context are explored.

3.
Food Res Int ; 167: 112688, 2023 05.
Artigo em Inglês | MEDLINE | ID: mdl-37087260

RESUMO

Ultra-processed, plant-based burgers (PB) and traditional comminuted-beef burgers (BB) share similar organoleptic characteristics, yet a knowledge gap exists in understanding how consumption of these divergent physical structures alters the lipemic response and gut microbiota. PB, comprised of highly refined ingredients, is formulated with no intact whole food structure, while BB entraps lipids throughout the myofibrillar protein network. PB presented significantly higher free fatty acid (FFA) bioaccessibility (28.2 ± 4.80 %) compared to BB (8.73 ± 0.52 %), as obtained from their FFA release profiles over digestion time after characterizing them with a modified logistic model (SLM), using the simulated TIM Gastro-Intestinal Model (TIM-1). Additionally, the rate of lipolysis, k, obtained from the SLM for PB (90% CI [0.0175, 0.0277] min-1) was higher than for BB (90% CI [0.0113, 0.0171] min-1). Using the Simulated Human Intestinal Microbial Ecosystem (SHIME®), the Firmicutes to Bacteroidetes ratio (F/B ratio) was significantly higher for PB than BB; and linear discriminant analysis effect size (LEfSe) showed Clostridium and Citrobacter were more highly represented in the microbial community for the PB feed, whereas BB feed differentially enriched Megasphaera, Bacteroides, Alistipes, and Blautia at the genus level. Additionally, short-chain fatty acid (SCFA) production was altered (p < 0.05) site-specifically in each colon vessel, which could be attributed to the available substrates and changes in microbial composition. Total SCFAs were significantly higher for PB in the ascending colon (AC) and descending colon (DC) but higher for BB only in the transverse colon (TC). This research illustrates the crucial role of meat analog physical structure in modulating nutritional aspects beyond food composition alone.


Assuntos
Ecossistema , Intestinos , Animais , Humanos , Bovinos , Fezes , Colo , Ácidos Graxos Voláteis , Bacteroidetes
4.
Foods ; 12(23)2023 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-38231609

RESUMO

This study investigated the mechanism of how lauric arginate ethyl ester (LAE) improves the photoinactivation of bacteria by curcumin after diluting the 100 µmol/L stock curcumin-LAE micelle solution to the concentration used during the treatment based on the curcumin concentration. The photoinactivation of bacteria was conducted by irradiating the 1 µmol/L curcumin-LAE solution containing cocktails of Escherichia coli and Listeria innocua strains (7 log CFU/mL) for 5 min with UV-A light (λ = 365 nm). The changes in solution turbidity, curcumin stability, and bacterial morphology, viability, and recovery were observed using SEM, TEM, and live/dead cell assays. The study found that LAE enhances the photoinactivation of bacteria by increasing the permeability of cell membranes which could promote the interaction of reactive oxygen species produced by photosensitized curcumin with the cell components. The combination of curcumin and LAE was demonstrated to be more effective in inhibiting bacterial recovery at pH 3.5 for E. coli, while LAE alone was more effective at pH 7.0 for L. innocua.

5.
Antioxidants (Basel) ; 11(11)2022 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-36358510

RESUMO

Lipid oxidation is a major pathway for the chemical deterioration of low-moisture foods. Little is known about how the physical properties of the fat used in crackers impact lipid oxidation kinetics. Fully hydrogenated soybean fat + interesterified soybean oil, fully hydrogenated soybean fat + sunflower oil, fully hydrogenated soybean oil, and soybean oil and interesterified fat alone were formulated to have varying solid fat content (SFC) at 55 °C but the same linoleic acid and tocopherol contents, so the fats had similar susceptibility to oxidation. A fluorescence probe showed that lipid mobility increased with decreasing SFC in both cracker doughs and fat blends, suggesting the probe could be used to monitor SFC directly in foods. Decreasing SFC decreased oxidation in crackers. Crackers made from interesterified fat (13.7% SFC) were more oxidatively stable (hexanal lag phase = 33 days) than crackers made from fat blends (hexanal lag phase = 24 days). These results suggest that blended fats result in regions of liquid oil high in unsaturated fatty acids within a food product prone to oxidation. Conversely, interesterified fats where unsaturated and saturated fatty acids are more evenly distributed on the triacylglycerols are more stable. Thus, interesterified fats could allow for the formulation of products higher in unsaturated fatty acids to improve nutritional profiles without sacrificing shelf life.

6.
Curr Res Food Sci ; 5: 479-490, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35265856

RESUMO

The volume-spanning network formed by gluten during breadmaking is crucial in the production of high-quality bakery products. Zein proteins are also capable of forming a protein network under specific conditions. Vibrational (Fourier transform infrared spectroscopy (FTIR) and Raman scattering) and fluorescence spectroscopy are powerful, non-invasive techniques capable of assessing protein structures and interactions. The main objective of this project was to explore the suitability of these techniques to study zein and gluten structures and interactions in complex dough systems. The dough samples were prepared by mixing 20 w/w% of protein (with different proportions of zein and gluten) and 80 w/w% of corn starch. The tyrosine (Tyr) fluorescence emission peak (λexc = 280 nm) was still present even in those zein-gluten samples containing the highest gluten concentration and lowest zein concentration. This suggests that the Tyr moieties (stemming from zein) are not in close proximity to tryptophan (Trp) of gluten and their fluorescence is not quenched efficiently. Raman scattering results also showed the presence of different Tyr residues, exposed and buried, as well as different conformations of disulfide bridges, in zein and gluten samples. Based on the results from spectroscopic measurements and scanning electron microscopy (SEM), two distinct network structures composed of gluten and zein were identified in the mixed dough systems. The present work illustrates how complementary vibrational (Raman scattering and FTIR) and fluorescence spectroscopy methods can be combined to non-invasively assess protein structure and interactions in a complex food matrix.

7.
Food Chem ; 382: 132266, 2022 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-35134719

RESUMO

C-Phycocyanin (C-PC) represents an alternative to artificial blue/green dyes in food products. This study characterized and gained insights into C-PC thermal stability mechanisms and provided a model to estimate its thermal degradation. Aqueous solutions of C-PC (0.3 µM, pH:6.1) were isothermally heated at 45-80 °C. C-PC degradation was monitored based on the photophysical properties of its lumiphores (phycocyanobilins and aromatic aminoacids-AAs). While C-PC was stable at 45 °C, less than 10 min at 80 °C sufficed to degrade most of it. The thermal degradation curves were characterized using the Weibull model, which was validated with data obtained under non-isothermal conditions. Deviations between estimated and experimental values were lower than 8%. Hypsochromic shifts of the AAs' spectra (from 340 to 315 nm) and increase (>30%) in anisotropy at λexc = 280 and 520 nm suggest that colour losses are not solely associated with alterations of the chromophore but also with conformational changes and possible aggregation of the protein subunits.


Assuntos
Temperatura Alta , Ficocianina , Cinética , Água
8.
Food Chem ; 376: 131928, 2021 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-34968903

RESUMO

The molecular mobility of amorphous excipients is important for the stability of biomaterials during preservation, facilitating matrix formulation and product design. Phosphorescence spectroscopy is a sensitive optical method to study molecular mobility. However, there is a need to expand the pool of probes available for analysis since molecules differ in sensitivity. This research explored the feasibility and limitations of using riboflavin as a phosphorescent probe for monitoring matrix molecular mobility. Phosphorescence decays of riboflavin in four amorphous cryosolvents (aqueous solutions of glycerol, ethanol, sucrose, and dextran) were collected at 77 K to capture its natural phosphorescence lifetime (estimated at 170 ms). Decays were also collected during ballistic heating to assess the sensitivity of riboflavin towards changes in matrix molecular mobility. Riboflavin exhibited good sensitivity towards matrix secondary relaxations in the glass, indicating that riboflavin has excellent potential as an edible phosphorescent probe for molecular mobility in food and pharmaceutical products.

9.
Curr Res Food Sci ; 4: 598-602, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34485928

RESUMO

The COVID-19 pandemic has generated increased interest in potential transmission routes. In food retail settings, transmission from infected customers and workers and customers through surfaces has been deemed plausible. However, limited information exists on the presence and survival of SARS-CoV-2 on surfaces, particularly outside laboratory settings. Therefore, the purpose of this project was to assess the presence of the virus at commonly found surfaces at food retail stores and the potential role that these spaces play in virus transmission. Samples (n=957) were collected twice a week for a month in food-retail stores within Ontario, Canada. High-touch surfaces were identified and surveyed in 4 zones within the store (payment stations, deli counters, refrigerated food section and carts and baskets). The samples were analyzed using a molecular method, i.e., reverse transcriptase quantitative Polymerase Chain Reaction (RT-qPCR). Regardless of the store's location, the sampling day or time, the location of the surface within the store or the surface material, all samples tested negative for SARS-CoV-2. These results suggest that the risk of exposure from contaminated high-touch surfaces within a food retailer store is low if preventive measures and recommended sanitizing routines are maintained.

10.
Foods ; 10(8)2021 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-34441554

RESUMO

Microbial photoinactivation using ultraviolet (UV) or visible light can be enhanced by photosensitizers. This study assessed the efficacy of encapsulating a food-grade photosensitizer (curcumin) in surfactant micelles on its water dispersibility, chemical stability, and antimicrobial activity. Stock curcumin-surfactant solutions were prepared with Surfynol 465 (S465) or Tween 80 (T80) (5 mM sodium citrate buffer). The antimicrobial activity of curcumin-loaded surfactant solutions was determined by monitoring the inactivation of Escherichia coli O157: H7 and Listeria innocua after 5-min irradiation with UV-A light (λ = 365 nm). The solutions mixed with the bacterial suspensions contained 1 µM curcumin and each surfactant below, near, and above their critical micelle concentrations (CMCs). The addition of surfactants at any level to the curcumin solution enhanced its dispersibility, stability, and efficacy as a photosensitizer, thereby enhancing its antimicrobial activity. Gram-positive bacteria were more susceptible than Gram-negative bacteria when curcumin-loaded micelles were used against them. The photoinactivation efficacy of curcumin-surfactant solutions depended on the pH of the solution (low > high), surfactant type (S465 > T80), and the amount of surfactant present (below CMC ≥ near CMC > above CMC = unencapsulated curcumin). This result suggests that excessive partitioning of curcumin into micelles reduced its ability to interact with microbial cells. Synergistic antimicrobial activity was observed when S465 was present below or near the CMC with curcumin at pH 3.5, which could be attributed to a more effective interaction of the photosensitizer with the cell membranes as supported by the fluorescence lifetime micrographs. The use of a micelle-based delivery system facilitates adsorption and generation of reactive oxygen species in the immediate environment of the microbial cell, enhancing photoinactivation.

11.
Heliyon ; 6(4): e03769, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32373726

RESUMO

The present study identified the threshold concentration of xylo-oligosaccharides (XOS) that resulted in minimal quality changes (rheology, color, water activity, pH, and total soluble solids) in strawberry puree. Optimization of XOS concentration to 5% (w/w) did not significantly alter the quality attributes of the strawberry puree. In addition, this study also monitored the rheological properties, composition (total soluble solids, total phenolic content, flavonoids, and tannin content), physicochemical attributes (color, water activity, pH) and sensorial properties of XOS-enhanced (5%, w/w) strawberry puree after thermal processing (HTST: 75 °C, 15s and UHT: 121 °C, 2s) and storage after 1, 15, and 36 days at 4 °C and 55 °C. At 5% (w/w) concentration, the addition of XOS increased consumer preference without significantly compromising quality attributes. Thermally treated strawberry puree (HTST and UHT) were less preferred by consumers than fresh puree. However, all strawberry samples incorporated with XOS (5%, w/w) received statistically higher scores than the samples without the XOS addition. Thus, the proposed supplementation of strawberry puree with XOS could be a viable solution to increase consumers' dietary fiber intake with little need for behavioral changes.

12.
Soft Matter ; 15(45): 9205-9214, 2019 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-31710326

RESUMO

Peptides are a promising class of gelators, due to their structural simplicity, biocompatibility and versatility. Peptides were synthesized based on four amino acids: leucine, phenylalanine, tyrosine and tryptophan. These peptide gelators, with systematic structural variances in side chain structure and chain length, were investigated using Hansen solubility parameters to clarify molecular features that promote gelation in a wide array of solvents. It is of utmost importance to combine both changes to structural motifs and solvent in simultaneous studies to obtain a global perspective of molecular gelation. It was found that cyclization of symmetric dipeptides, into 2,5-diketopiperazines, drastically altered the gelation ability of the dipeptides. C-l-LL and C-l-YY, which are among the smallest peptide LMOGs reported to date, are robust gelators with a large radius of gelation (13.44 MPa1/2 and 13.90 MPa1/2, respectively), and even outperformed l-FF (5.61 MPa1/2). Interestingly, both linear dipeptides (l-FF and l-LL) gelled similar solvents, yet when cyclized only cyclo-dityrosine was a robust gelator, while cyclo-diphenylalanine was not. Changes in the side chains drastically affected the crystal morphology of the resultant gels. Symmetric cyclo dipeptides of leucine and tyrosine were capable of forming extremely high aspect ratio fibers in numerous solvents, which represent new molecular motifs capable of driving self-assembly.


Assuntos
Peptídeos/química , Motivos de Aminoácidos , Sequência de Aminoácidos , Ciclização , Géis , Solubilidade
13.
J Colloid Interface Sci ; 556: 568-576, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31479830

RESUMO

The objective of this study was to provide insights into the mechanisms involved in the mass transport of antimicrobial compounds from essential oil nanoemulsions to bacterial cell membranes. Origanum oil-in-water nanoemulsions were produced using spontaneous emulsification by titrating a mixture of essential oil, ripening inhibitor, and surfactant (Tween 80) into 5 mM sodium citrate buffer (pH 3.5). Stable nanoemulsions containing relatively small droplets (d < 60 nm) were produced using this low-energy method. The nature of the ripening inhibitor used in the oil phase of the nanoemulsions affected the antimicrobial activity of the nanoemulsions: corn (LCT) > medium-chain triglycerides (MCT). Differences in antimicrobial activity were attributed to the differences in the rate of transfer of hydrophobic antimicrobial constituents from the nanoemulsion to the MCT emulsion, which was used to mimic the hydrophobic region of the bacterial cell membranes. Each antimicrobial nanoemulsion was separated from the MCT emulsion by a dialysis tubing. Dialysis tubing with two different pore sizes was used, one excluding nanoemulsion droplet and micelle delivery, allowing the delivery of antimicrobial compounds only through the aqueous phase and the other by both the aqueous phase and micelles. For origanum oil nanoemulsions, the delivery of all antimicrobial agents occurred more efficiently when micelles were present.


Assuntos
Anti-Infecciosos , Nanoestruturas/química , Óleos Voláteis , Polissorbatos , Salmonella/crescimento & desenvolvimento , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Parede Celular/metabolismo , Emulsões , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Polissorbatos/química , Polissorbatos/farmacologia
14.
Food Sci Nutr ; 7(2): 506-518, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30847129

RESUMO

Lowering the sodium content in meat products, particularly in beef patties, can be challenging because sodium plays many functional roles in these products. Meat extenders can contribute to lower sodium content by imparting complementary flavors while reducing caloric and sodium content. A systematic comparison of two meat extenders, namely mushrooms and textured soy (TSP) in terms of physical and sensory characteristics, is presented herein. The physical properties of the samples suggested that the use of mushroom and TSP extender would perform statistically similar to an all-meat control depending on the level of substitution. Hedonic sensory analysis showed meat extension using mushrooms yielded liking scores more similar to the all-meat formulations than TSP in reduced sodium applications. The results of this research suggest that mushrooms have the potential to be successfully incorporated into reduced sodium meat products to provide a healthier product.

15.
J Funct Foods ; 45: 268-276, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30416540

RESUMO

Malanga (Xanthosoma sagittifolium) is used as a medicinal food for infant development and gastritis. We compared the physicochemical properties and gut microbial effects of malanga versus potato (Solanum tuberosum) using nutritional analysis, rheometry, in vitro TNO Intestinal Model, and C57Bl/6J mouse models. Malanga was characterized by higher starch (70.7% v. 66.3%), lower amylose:amylopectin (0.33 v. 0.59), higher free sugar (5.44% v. 3.23%), lower viscosity (271.0 v. 863.0 mPa.s), and higher bioaccessible and bioavailable sugar (0.89 v. 0.11 g bioaccessible sucrose per 20 g load in vitro; blood glucose levels of 129.1 v. 95.2 and 133.8 v. 104.3 mg/dL after 20 and 60 min in vivo). Gut microbiota of mice fed a high fat diet containing 20% malanga for 14 d exhibited significantly higher α diversity than those fed 20% potato, indicating that minor physicochemical differences between similar tuber crops are associated with significantly different effects on the gut microbiome.

16.
Annu Rev Food Sci Technol ; 9: 251-269, 2018 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-29328810

RESUMO

The labels currently used on food and beverage products only provide consumers with a rough guide to their expected shelf lives because they assume that a product only experiences a limited range of predefined handling and storage conditions. These static labels do not take into consideration conditions that might shorten a product's shelf life (such as temperature abuse), which can lead to problems associated with food safety and waste. Advances in shelf-life estimation have the potential to improve the safety, reliability, and sustainability of the food supply. Selection of appropriate kinetic models and data-analysis techniques is essential to predict shelf life, to account for variability in environmental conditions, and to allow real-time monitoring. Novel analytical tools to determine safety and quality attributes in situ coupled with modern tracking technologies and appropriate predictive tools have the potential to provide accurate estimations of the remaining shelf life of a food product in real time. This review summarizes the necessary steps to attain a transition from open labeling to real-time shelf-life measurements.


Assuntos
Armazenamento de Alimentos , Inocuidade dos Alimentos , Modelos Estatísticos , Fatores de Tempo
17.
Crit Rev Food Sci Nutr ; 58(11): 1902-1916, 2018 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-28662357

RESUMO

Fluorescent molecular rotors (MRs) are compounds whose emission is modulated by segmental mobility; photoexcitation generates a locally excited (LE), planar state that can relax either by radiative decay (emission of a photon) or by formation of a twisted intramolecular charge transfer (TICT) state that can relax nonradiatively due to internal rotation. If the local environment around the probe allows for rapid internal rotation in the excited state, fast non-radiative decay can either effectively quench the fluorescence or generate a second, red-shifted emission band. Conversely, any environmental restriction to twisting in the excited state due to free volume, crowding or viscosity, slows rotational relaxation and promotes fluorescence emission from the LE state. The environmental sensitivity of MRs has been exploited extensively in biological applications to sense microviscosity in biofluids, the stability and physical state of biomembranes, and conformational changes in macromolecules. The application of MRs in food research, however, has been only marginally explored. In this review, we summarize the main characteristics of fluorescent MRs, their current applications in biological research and their current and potential applications as sensors of physical properties in food science and engineering.


Assuntos
Corantes Fluorescentes/química , Tecnologia de Alimentos , Fluorescência , Interações Hidrofóbicas e Hidrofílicas , Sondas Moleculares/química , Estrutura Molecular , Conformação Proteica , Dobramento de Proteína , Proteínas/química , Proteólise , Relação Estrutura-Atividade , Viscosidade
18.
Food Chem ; 245: 104-111, 2018 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-29287320

RESUMO

The objective of this research was to study the impact of ripening inhibitor level and type on the formation, stability, and activity of antimicrobial thyme oil nanoemulsions formed by spontaneous emulsification. Oil-in-water antimicrobial nanoemulsions (10 wt%) were formed by titrating a mixture of essential oil, ripening inhibitor, and surfactant (Tween 80) into 5 mM sodium citrate buffer (pH 3.5). Stable nanoemulsions containing small droplets (d < 70 nm) were formed. The antimicrobial activity of the nanoemulsions decreased with increasing ripening inhibitor concentration which was attributed to a reduction in the amount of hydrophobic antimicrobial constituents transferred to the separated hydrophobic domain, mimicking bacterial cell membranes, by using dialysis and chromatography. The antimicrobial activity of the nanoemulsions also depended on the nature of the ripening inhibitor used: palm ≈ corn > canola > coconut which also depended on their ability to transfer hydrophobic antimicrobial constituents to the separated hydrophobic domain.


Assuntos
Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Thymus (Planta)/química , Triglicerídeos/química , Bactérias/química , Bactérias/efeitos dos fármacos , Membrana Celular , Diálise , Emulsões/química , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Nanoestruturas/química , Óleos Voláteis/química , Polissorbatos/química , Tensoativos/química
19.
Langmuir ; 33(41): 10907-10916, 2017 10 17.
Artigo em Inglês | MEDLINE | ID: mdl-28926708

RESUMO

1,3:2,4-Dibenzylidene-d-sorbitol (DBS) is the gold-standard for low-molecular-weight organogelators (LMOGs). DBS gels a wide array of solvents, as illustrated by the large Hansen sphere representing gels (2δd = 33.5 MPa1/2, δp = 7.5 MPa1/2, and δh = 8.7 MPa1/2; radius = 11.2 MPa1/2). Derivatives of DBS have been synthesized to isolate and determine molecular features essential for organogelation. In this work, π-π stacking and hydrogen bonding are the major noncovalent interactions examined. The importance of π-π stacking was studied using 1,3:2,4 dicyclohexanecarboxylidene-d-sorbitol (DCHS), which eliminates possible π-π stacking while still conserving the other structural aspects of DBS. The replacement of the benzyl groups with cyclohexyl groups led to a very a poor gelator; only one of the several solvents examined, carbon tetrachloride, formed a gel. 1,3:2,4-Diethylidene-d-sorbitol (DES), another DBS analogue incapable of π-π stacking but with very different polarity, gelated a large Hansen space (2δd = 34.0 MPa1/2, δp = 10.9 MPa1/2, and δh = 10.8 MPa1/2; radius = 9.2 MPa1/2). DES gels solvents with higher δp and δh values than DBS. To assess the role of hydrogen bonding, DBS was acetalated (A-DBS), and it was found that the Hansen space gelated by A-DBS shifted to less polar solvents with higher hydrogen-bonding Hansen solubility parameters (HSPs) (2δd = 33.8 MPa1/2, δp = 6.3 MPa1/2, and δh = 9.6 MPa1/2; radius = 11.1 MPa1/2) than for DBS. These systematic structural modifications are the first step in exploring how specific intermolecular features alter aspects of Hansen space corresponding to positive gelation outcomes.

20.
J Fluoresc ; 27(5): 1621-1631, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28432632

RESUMO

Five variants of glucokinase (ATP-D-hexose-6-phosphotransferase, EC 2.7.1.1) including wild type and single Trp mutants with the Trp residue at positions 65, 99, 167 and 257 were prepared. The fluorescence of Trp in all locations studied showed intensity changes when glucose bound, indicating that conformational change occurs globally over the entire protein. While the fluorescence quantum yield changes upon glucose binding, the enzyme's absorption spectra, emission spectra and fluorescence lifetimes change very little. These results are consistent with the existence of a dark complex for excited state Trp. Addition of glycerol, L-glucose, sucrose, or trehalose increases the binding affinity of glucose to the enzyme and increases fluorescence intensity. The effect of these osmolytes is thought to shift the protein conformation to a condensed, high affinity form. Based upon these results, we consider the nature of quenching of the Trp excited state. Amide groups are known to quench indole fluorescence and amides of the polypeptide chain make interact with excited state Trp in the relatively unstructured, glucose-free enzyme. Also, removal of water around the aromatic ring by addition of glucose substrate or osmolyte may reduce the quenching.


Assuntos
Fluorescência , Glucoquinase/química , Conformação Proteica , Triptofano/química , Glucoquinase/genética , Glucoquinase/metabolismo , Humanos , Mutação , Espectrometria de Fluorescência , Especificidade por Substrato , Triptofano/metabolismo
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