Dipolar order in an amphidynamic crystalline metal-organic framework through reorienting linkers.

Amphidynamic crystals, which possess crystallinity and support dynamic behaviours, are very well suited to the exploration of emergent phenomena that result from the coupling on the dynamic moieties. Here, dipolar rotors have been embedded in a crystalline metal-organic framework. The material consists of Zn(II) nodes and two types of ditopic bicyclo[2.2.2]octane-based linkers-one that coordinates to the Zn clusters through two 1,4-aza moieties, and a difluoro-functionalized derivative (the dipolar rotor) that coordinates through linked 1,4-dicarboxylate groups instead. Upon cooling, these linkers collectively order as a result of correlated dipole-dipole interactions. Variable-temperature, frequency-dependent dielectric measurements revealed a transition temperature Tc = 100 K, when a rapidly rotating, dipole-disordered, paraelectric phase transformed into an ordered, antiferroelectric one in which the dipole moments of the rotating linkers largely cancelled each other. Monte Carlo simulations on a two-dimensional rotary lattice showed a ground state with an Ising symmetry and the effects of dipole-lattice and dipole-dipole interactions.

Proteolytic System of Streptococcus thermophilus.

The growth of lactic acid bacteria (LAB) generates a high number of metabolites related to aromas and flavors in fermented dairy foods. These microbial proteases are involved in protein hydrolysis that produces necessary peptides for their growth and releases different molecules of interest, like bioactive peptides, during their activity. Each genus in particular has its own proteolytic system to hydrolyze the necessary proteins to meet its requirements. This review aims to highlight the differences between the proteolytic systems of Streptococcus thermophilus and other lactic acid bacteria (Lactococcus and Lactobacillus) since they are microorganisms that are frequently used in combination with other LAB in the elaboration of fermented dairy products. Based on genetic studies and in vitro and in vivo tests, the proteolytic system of Streptococcus thermophilus has been divided into three parts: 1) a serine proteinase linked to the cellular wall that is activated in the absence of glutamine and methionine; 2) the transport of peptides and oligopeptides, which are integrated in both the Dpp system and the Ami system, respectively; according to this, it is worth mentioning that the Ami system is able to transport peptides with up to 23 amino acids while the Opp system of Lactococcus or Lactobacillus transports chains with less than 13 amino acids; and finally, 3) peptide hydrolysis by intracellular peptidases, including a group of three exclusive of S. thermophilus capable of releasing either aromatic amino acids or peptides with aromatic amino acids.

Probiotic Lactobacillus Strains Stimulate the Inflammatory Response and Activate Human Macrophages.

Lactobacilli have been shown to promote health functions. In this study, we analyzed the mechanism by which four different strains of probiotics affected innate immunity, such as regulation of ROS, cytokines, phagocytosis, bactericidal activity, signaling by NF-κB pp65, and TLR2 activation. The production of ROS was dependent on the concentration and species of Lactobacillus. The results obtained from the tested strains (Lactobacillus rhamnosus GG, L. rhamnosus KLSD, L. helveticus IMAU70129, and L. casei IMAU60214) showed that strains induced early proinflammatory cytokines such as IL-8,TNF-α, IL-12p70, and IL-6. However, IL-1ß expression was induced only by L. helveticus and L. casei strains (after 24 h stimulation). Phagocytosis and bactericidal activity of macrophages against various pathogens, such as S. aureus, S. typhimurium, and E. coli, were increased by pretreatment with Lactobacillus. The nuclear translocation NF-κB pp65 and TLR2-dependent signaling were also increased by treatment with the probiotics. Taken together, the experiments demonstrate that probiotic strains of Lactobacillus exert early immunostimulatory effects that may be directly linked to the initial inflammation of the response of human macrophages.

Enhancement of lactase activity in milk by reactive sulfhydryl groups induced by heat treatment.

The effects of heat treatments of milk and whey prior to lactose hydrolysis with Kluyveromyces lactis beta-galactosidase were studied. It was observed that heat treatment of milk significantly increases lactase activity, with a maximum activity increase found when milk was heated at 55 degrees C. In whey from 55 up to 75 degrees C, beta-galactosidase activity decreased slightly. Nevertheless, heating whey at 85 degrees C for 30 min raised the rate of hydrolysis significantly. Electrophoretic patterns and UV spectra proved that the activity change correlated with milk protein denaturation, particularly that of beta-lactoglobulin. Heating whey permeate did not increase the enzyme activity as heating whole whey; but heating whey prior to ultrafiltration also resulted in enzyme activation. Measurement of free sulfhydryl (SH) groups in both whey and heated whey permeate showed that the liberation of free SH is highly correlated to the change of the activity. Furthermore, this activation can be reversed by oxidizing the reactive sulfhydryl groups, proving that the observed effect may be related to the release of free SH to the medium, rather than to the denaturation of a thermolabile protein inhibitor.

Steps to demarcate the effects of chromophore aggregation and planarization in poly(phenyleneethynylene)s. 1. Rotationally interrupted conjugation in the excited states of 1,4-bis(phenylethynyl)benzene.

A series of photophysical measurements and semiempirical calculations were carried out with 1,4-bis(phenylethynyl)benzene in search of evidence on the effects of phenyl group rotation and chromophore aggregation of oligo- and poly(phenyleneethynylene)s. It is suggested that planarization gives rise to relatively modest shifts of ca. 20-30 nm, which preserve the vibronic structure of the monomer and retain a high emission quantum yield. In contrast, it is proposed that aggregation gives rise to larger shifts and loss of vibronic structure.

Engineering reactions in crystalline solids: photochemical generation of secondary and tertiary enol radical pairs from crystalline ketodiesters.

The photochemical decarbonylation of several crystalline 1,3-acetonedicarboxylates has been analyzed in solution and in the solid state. It is shown that the efficiency of the solid-state reaction depends on the stability of the intermediate acyl-alkyl and alkyl-alkyl radical pairs. Reactions proceeding through tertiary enol radicals are more efficient than reactions proceeding through secondary enol radical centers. Solid-state reactions that require the intermediacy of primary enol radicals do not occur. It is also shown that the selectivity of product formation in crystals depends on the structure of the reactant solid phase.

Steps to demarcate the effects of chromophore aggregation and planarization in poly(phenyleneethynylene)s. 2. The photophysics of 1,4-diethynyl-2-fluorobenzene in solution and in crystals.

Crystals of 1,4-bis(2-hydroxy-2-methyl-3-butynyl)-2-fluorobenzene 4 have a rich packing structure with four distinct molecules in the unit cell. A complex hydrogen bonding network results in the formation of cofacial trimers, cofacial dimers, and monomers within the same unit cell. Given a remarkable opportunity to investigate the effect of aggregation on the photophysics of 1,4-diethynylbenzenes, we analyzed the absorption, diffuse reflectance, and emission spectra of compound 4 in solutions and in crystals. Diffuse reflectance and fluorescence excitation revealed a red-shifted absorption that is absent in dilute solution but becomes observable at high concentrations and low temperatures. The fluorescence emission in the solid state is dual with components assigned to monomers and aggregates. The excitation and emission assigned to the monomer are nearly identical in crystals and dilute solutions. The absorption and emission bands assigned to aggregates are broad and red-shifted by 60--80 nm. As expected for a sample with absorbers and emitters with different energies and incomplete equilibration, efficient monomer-to-aggregate energy transfer was observed by a proper selection of excitation wavelengths. The fluorescence quantum yield of 4 in solution is relatively low (Phi(F) = 0.15) and the singlet lifetime short (tau(F) = 3.8 ns). A lower limit for the triplet yield of Phi(T) = 0.64 was determined indirectly in solution by (1)O(2) sensitization, and a relatively strong and long-lived phosphorescence was observed in low-temperature glasses and in crystals at 77 K.

Effect of beta-galactosidase hydration on alcoholysis reaction in organic one-phase liquid systems.

Alcoholysis reactions were performed in organic one-phase liquid systems with E. coli beta-galactosidase to produce heptyl-beta-galactoside from lactose and 1-heptanol. The reaction rate was highly dependent on the amount of water solubilized in the alcohol. A larger amount of water led to a system of two liquid phases in which the alcoholysis rate was 73% faster than in the one-phase system. No hydrolysis reaction of either lactose or product was observed in one-phase liquid systems up to 20 h, independent of the water content. Solubility of lactose in the organic phase increased with the water content in the system and the reaction followed the Michaelis-Menten model. Water activity was calculated for heptanol containing different amounts of water and the obtained values were used to estimate the hydration of beta-galactosidase from known models. Enzyme activity correlated with sorbed water, similar to the behavior reported for lysozyme in low water environments. It is concluded that water contribution to enzyme hydration dominates the rate of reaction compared to its effect on lactose solubilization.

Alcoholysis and reverse hydrolysis reactions in organic one-phase system with a hyperthermophilic beta-glycosidase.

Alcoholysis and reverse hydrolysis reactions were performed enzymatically in one-phase water-saturated 1-heptanol systems. Lactose or glucose was used as substrate to produce heptyl-beta-galactoside and/or heptyl-beta-glucoside, respectively. When alcoholysis of lactose was performed at 37 degrees C with beta-galactosidase from Escherichia coli, the initial rate was 14 nmol/mL min, and the limiting factors were the poor solubility of the substrate in 1-heptanol and low thermal stability of the enzyme. When a hyperthermophilic beta-glycosidase was used at 90 degrees C, the rate was 3.14-fold higher; in this case a higher concentration of soluble lactose in the water-saturated heptanol was available to the enzyme due to the higher temperature. The hyperthermophilic beta-glycosidase was also able to use glucose and galactose as substrates to achieve the reverse hydrolysis reaction. As a consequence, when lactose was used as substrate, heptyl-beta-galactoside was formed by alcoholysis, while the released glucose moiety was used in a secondary reverse hydrolysis reaction to produce heptyl-beta-glucoside. Both reactions followed Michaelis-Menten kinetics behavior. Neither lactose nor heptyl glycosides were hydrolyzed by this enzyme in water-saturated heptanol. However, the conversion was limited by a strong product inhibition and the formation of oligosaccharides, especially at high substrate concentrations, reducing the final glycoside yield.

[Biosynthesis of congeners during alcohol fermentation]. / Biosíntesis de congenéricos durante las fermentaciones alcohólicas.

The flavor of alcoholic beverages is a consequence of a complex mixture of many compounds, including small concentrations of some volatile metabolites known as congeners, which are produced by the yeast during the fermentation. The more important compounds are those that can be found in all the alcoholic beverages in different concentrations, and they can be grouped on the following chemical species: higher alcohols, esters, and carboniles. In the current paper the biochemical pathways that produce these compounds from the raw materials are reviewed. Research done in this field has led to a more complete knowledge concerning to organoleptic profiles of alcoholic beverages and to a better control for the production of the final product.

Enzymes involved in carbohydrate metabolism and their role on exopolysaccharide production in Streptococcus thermophilus.

The role of the enzymes uridine-5'-diphospho-(UDP) glucose pyrophosphorylase and UDP galactose 4-epimerase in exopolysaccharide production of Gal- ropy and non-ropy strains of Streptococcus thermophilus in a batch culture was investigated. Growth of the ropy and non-ropy strains was accompanied by total release of the galactose moiety from lactose hydrolysis in modified Bellinker broth with lactose as the only carbon source. This was associated with a greater exopolysaccharide production by the ropy strain. The polymer produced by both strains in cultures with lactose or glucose as carbon sources contained glucose, galactose and rhamnose, indicating that glucose was used as a carbon source for bacterial growth and for exopolysaccharide formation. UDP-glucose pyrophosphorylase activity was associated with polysaccharide production during the first 12 h in a 20 h culture in the ropy strain, but not in the non-ropy strain. UDP-galactose 4-epimerase was not associated with exopolysaccharide synthesis in any strain. The evidence presented suggests that the glucose moiety from lactose hydrolysis is the source of sugar for heteropolysaccharide synthesis, due to a high UDP-glucose pyrophosphorylase activity.

[Uses of microbial beta-galactosidases to reduce lactose content in milk and dairy products]. / Usos de beta-galactosidasas microbianas para reducir el contenido de lactosa en leche y productos lácteos.

The commercial sources of microbial beta-galactosidases (lactases) include the yeasts species Kluyveromyces marxianus, Kluyveromyces lactis and Candida kefyr which are used to hydrolyse lactose in milk due to their optimum pH. On the other hand, lactases obtained from the moulds Aspergillus niger and Aspergillus oryzae have an acid optimum pH and therefore are used to hydrolyse lactose in whey to obtain whey syrups to be used as raw materials in the food industry. The lactose intolerance problem has led to many studies concerning lactose hydrolysis by means of these microbial enzymes to obtain milk suitable for people with lactose maldigestion and special diets for ill persons, elderly population and intolerant babies due to secondary deficiency of lactose. Most industries obtain hydrolyzed lactose milk with free enzyme; however, there are some developments of immobilized lactase catalysts which are being used mainly in whey.

Chemical composition of a mixture of single-cell protein obtained from Kluyveromyces fragilis and whey proteins.

A mixture of Kluyveromyces fragilis biomass and coagulated whey proteins was obtained by fermentation of whole whey. This product had a chemical composition similar to that of washed products reported in the literature with high-crude protein and low-ash contents. The product had a high content of sulphur-containing amino acids and tryptophan, which are usually limiting in yeast biomass. Lysine content was inexplicably lower than the expected value, being the limiting amino acid in this case. The chemical score of the protein was 91%. From the biomass-whey proteins product a protein isolate could be recovered with a yield of 80%. The protein content of the isolate was 75%, and the nucleic acids were reduced by 90.8%. The cell-wall debris were also considerably reduced.