Flavor Improvement of Maillard Reaction Intermediates Derived from Enzymatic Hydrolysates of Oudemansiella raphanipes Mushroom.

Maillard reaction intermediate (MRI) was prepared by the enzymatic hydrolysate (EH) of Oudemansiella raphanipes and fructose. The optimal preparation condition of MRIs was obtained when the Maillard reaction parameters were as follows: fructose addition of 5%, reaction time of 60 min, and temperature of 60 °C. E-Tongue results indicated that the umami and saltiness of MRIs were greater than those of Maillard reaction products (MRPs) and EH, and the taste-enhancing ability of MRIs was even more prominent than that of MRPs. E-Nose could obviously distinguish EH, MRIs, and MRPs, and there was an obvious difference between MRPs and MRIs regarding volatile aroma compounds. A total of 35 volatile flavor substances were identified among the three samples, including 6 alcohols, 13 aldehydes, 9 ketones, 2 esters, and 5 other compounds. Overall, MRIs could avoid the production of complete reaction products with an inferior flavor, and further enhance the umami taste.

Study on the Optimization, Extraction Kinetics and Thermodynamics of the Ultrasound-Assisted Enzymatic Extraction of Tremella fuciformis Polysaccharides.

In this study, Tremella fuciformis polysaccharides (TFPs) were extracted by ultrasound-assisted enzymatic extraction (UAE) at different extraction parameters in order to explore the potential of ultrasound in intensifying the extraction yield. The effects of experimental conditions on the extraction yields were optimized using response surface methodology, with the optimal ultrasonic power of 700 W, temperature of 45 °C and time of 50 min. The kinetic analysis revealed that UAE significantly promoted the dissolution, diffusion and migration with the maximum yield of 26.39%, which was enhanced by 40.45% and 156.96% compared with individual ultrasonic extraction (UE) and enzymatic extraction (EE). According to the modified Fick's second law of diffusion, the extraction process of TFPs illustrated a good linear correlation (R2 ≥ 0.9), and the rate constant gradually elevated as the temperature increased from 25 to 45 °C, while the presence of ultrasound exerted a vital role in extracting TFPs. Regarding to the thermodynamic results, the positive values of ΔH and ΔG demonstrated that UAE, UE and EE were endothermic and unspontaneous processes. This study provides a theoretical basis for polysaccharide extraction processing.

Sono-activation of food enzymes: From principles to practice.

Over the last decade, sono-activation of enzymes as an emerging research area has received considerable attention from food researchers. This kind of relatively new application of ultrasound has demonstrated promising potential in facilitating the modern food industry by broadening the application of various food enzymes, improving relevant industrial unit operation and productivity, as well as increasing the yield of target products. This review aims to provide insight into the fundamental principles and possible industrialization strategies of the sono-activation of food enzymes to facilitate its commercialization. This review first provides an overview of ultrasound application in the activation of food protease, carbohydrase, and lipase. Then, the recent development on ultrasound activation of food enzymes is discussed on aspects including mechanisms, influencing factors, modification effects, and its applications in real food systems for free and immobilized enzymes. Despite the far fewer studies on sono-activation of immobilized enzymes compared with those on free enzymes, we endeavored to summarize the relevant aspects in three stages: ultrasound pretreatment of free enzyme/carrier, assistance in immobilization process, and modification of the already immobilized enzyme. Lastly, challenges for the scalability of ultrasound in these target areas are discussed and future research prospects are proposed.

Insights into the relationships of modifying methods, structure, functional properties and applications of chitin: A review.

Chitin as the second plentiful polysaccharide has arouse widely attention due to its remarkable availability and biocompatibility. While the strong inter/intra molecular hydrogen bonds and crystallinity severely restrict its applications. Recently, multiple emerging technologies are increasingly used to modify chitin structure for the sake of obtaining excellent functional properties, as well as broadening the corresponding applications. Firstly, this review systematically outlines the features of single and combined methods for chitin modification. Then, the impacts of various modifying methods on the structural characteristics of chitin, including molecular weight, degree of acetylation and functional groups, are further summarized. In addition, the effects of these structural characteristics on the functional properties as well as its potential related applications are illustrated. The conclusion of this review provides better understanding of the relationships among the modifying methods, structure, properties and applications, contributing to chitin modification for the targeted purpose in the future study.

Characterization of Physicochemical Properties of Oil-in-Water Emulsions Stabilized by Tremella fuciformis Polysaccharides.

In this paper, emulsions stabilized by Tremella fuciformis polysaccharides (TFP) were prepared and the physiochemical properties were assessed. Results showed that the TFP emulsions illustrated the highest emulsifying activity (EAI) and emulsifying stability (ESI) when the concentration of TFP and oil were 0.8% and 10% (wt%). The higher pH value was in favor of the emulsifying properties, while the addition of NaCl impaired the stability, and the greater the concentration, the lower the EAI and ESI. Besides, the emulsifying and rheological properties and stability analysis were evaluated in comparison with gum arabic, pectin, and carboxymethyl cellulose emulsions. It was discovered that TFP illustrated better storage and freeze-thaw stability, which was proved by the result of zeta-potential and particle size. The rheological measurement revealed that all the emulsions behaved as pseudoplastic fluids, while TFP displayed a higher viscosity. Meanwhile, TFP emulsions tended to form a more stable network structure according to the analysis of the parameters obtained from the Herschel-Bulkley model. FTIR spectra suggested that the O-H bond could be destructed without the formation of new covalent bonds during the emulsion preparation. Therefore, this study would be of great importance for the research of emulsions stabilized by TFP as a natural food emulsifier.

Enhanced adsorption of Congo red using chitin suspension after sonoenzymolysis.

In the present work, chitin suspensions after enzymolysis and sonoenzymolysis were taken as adsorbents to evaluate the adsorption properties of Congo red (CR) dyes. Compared with untreated chitin suspension, the CR adsorption performance was significantly improved after enzymolysis and even more after sonoenzymolysis. According to different adsorption kinetic and isotherm models, Langmuir isotherm and the pseudo-second order model were more reliable to describe the adsorption process of CR onto different chitin samples and demonstrated a monolayer and favorable physisorption process. What's more, negative values of ΔG (Gibbs free energy change) and the shifts to higher negative values with the temperature increasing from adsorption thermodynamic study proved a spontaneous CR adsorption process. The structural characterization before and after adsorption further verified the physical adsorption between chitin and CR, and a larger specific area and higher porosity of chitin suspension was obtained after sonoenzymolysis with more available active sites.

Ultrasound enhanced the binding ability of chitinase onto chitin: From an AFM insight.

In order to evaluate the effect of ultrasound to chitinase from a molecular level, atomic force microscopy (AFM) was employed to investigate the interaction force of chitinase binding onto chitin surface. In the measurement of force-distance curve, a series of pull-off events were discovered using the immobilized AFM tips with chitinase either treated by ultrasound or not, whereas no interaction peak was observed by the AFM tips without chitinase, indicating that the obtained adhesion forces were coming from the binding functions between chitinase and chitin. Through the analysis of these force curves, at the loading velocity of 0.3 µm/s, the maximum binding force of the chitinase treated by ultrasound for 20 min onto chitin was measured to be 105.33 ± 23.51 pN, while the untreated onto chitin was 71.05 ± 12.73 pN, suggesting the stronger binding force between ultrasonic treated chitinase and chitin substrate. Therefore, AFM has provided a useful method to directly and quantitatively characterize the interactions between chitinase and chitin, and successfully proved that ultrasound could activate chitinase by enhancing the binding ability of chitinase onto chitin.

Proteomic responses of spores of Bacillus subtilis to thermosonication involve large-scale alterations in metabolic pathways.

Thermosonication (TS) impacts numerous characteristics of spores, such as morphology, cell metabolism, and stress resistance. However, relevant mechanisms need to be clarified. In the present study, the effect of TS treatment on Bacillus subtilis spores was investigated at phenotypic and proteomic levels. The results showed that TS treatment induced significant changes to spores in growth kinetics and morphology. A total of 167 differentially expressed proteins (DEPs) were obtained after TS treatment at 6.67 W/mL and 80 °C. Among these proteins, 80 were up-regulated, whereas 87 were down-regulated. These DEPs were classed into 20 functional categories. Enrichment analysis of the proteome revealed that the major categories were associated with metabolic functions, including energy metabolic processes, amino acids biosynthesis and metabolism, translation and ribosomal protein. In summary, B. subtilis spores showed alteration primarily in the proteins that were associated with metabolism under TS treatment. These findings could be applied to the development and optimization of TS-based sporicidal treatment.

Study on the mechanism of ultrasound-accelerated enzymatic hydrolysis of starch: Analysis of ultrasound effect on different objects.

Enzymatic hydrolysis of starch is an important process in the food industry. In the present work, ultrasound was introduced in glucoamylase pretreatment, starch pretreatment and mixed reaction system treatment to enhance starch hydrolysis efficiency. These different processes were studied to explore the mechanism of ultrasound in promoting enzymatic reactions. The hydrolysis degree of starch was determined via measuring the reducing sugar yield. Ultrasound caused enzyme inactivation under high temperatures, high ultrasonic power and long-time treatment, especially at high temperatures exceeding 65 °C. Ultrasound pretreatment of starch before enzymolysis led to the furtherance of starch hydrolysis degree. Meanwhile, sonicating the mixed enzymatic reaction system below 65 °C promoted starch hydrolysis significantly, inducing more than five- fold growth in the degree of starch hydrolysis as much as the ultrasound pretreatment caused. Molecular weights analysis conducted by the MALLS system reflected the enormous damage of starch molecules caused by ultrasound. The amylose contents and chain length distributions of samples were separately analyzed by iodine binding method and size exclusion chromatography. The results of the two experiments illustrate that ultrasound could promote the enzymatic hydrolysis of amylopectin, which is harder for glucoamylase to hydrolyze compared to amylose.

Enhancement of chitin suspension hydrolysis by a combination of ultrasound and chitinase.

This study evaluated the degradation kinetics and structural characteristics of chitin suspension (CS) with a combination of ultrasound and chitinase. Compared with the enzymolysis, the degradation degree of sonoenzymolysis was enhanced to the maximum by 27.93 % at an intensity of 25 W/mL for 20 min. According to degradation kinetics, ultrasound intensified molecular collision rate between chitinase and substrate, thereby increasing the degradation degree. What's more, combined with chitinase, ultrasound intensified the rate of the breakage of glycosidic bond and viscosity-average molecular weight (Mv) decrease, but no obvious change in acetylation degree (DA). Additionally, the intra- or inter-hydrogen bindings were weakened by ultrasound during sonoenzymolysis, leading to a slight decrease in crystalline index and a more ordered structure, which increased the accessibility of the substrate to enzyme. In conclusion, combination of chitinase and ultrasound could enhance the hydrolysis of CS while without changing its primary structure.

Comparison of citrus pectin and apple pectin in conjugation with soy protein isolate (SPI) under controlled dry-heating conditions.

In this study, we selected two most commonly-available commercial pectin, i.e. citrus pectin and apple pectin as the grafting polysaccharides to prepare soy protein isolate-pectin conjugates. Despite the similar degrees of methoxylation and acetylation for two pectin samples, apple pectin showed much more complex structures compared to citrus pectin, with a 2.20-fold higher molecular weight and large numbers of side chains. The conjugates were prepared under controlled dry-heating conditions and achieved the degree of graft of 25.00% and 21.85% for citrus and apple pectin, respectively. Formation of the conjugates was further confirmed by SDS-PAGE gel electrophoresis and IR spectra. Attributed to the strong steric-hindrance effect of pectin, the fluorescence intensity and surface hydrophobicity of the soy protein isolate were significantly decreased after Maillard reaction. However, both solubility and emulsifying properties of the conjugates were significantly improved. Results indicated that both pectin samples played favorable roles in protein modification.

Ultrasound pretreatment enhances the inhibitory effects of nisin/carvacrol against germination, outgrowth and vegetative growth of spores of Bacillus subtilis ATCC6633 in laboratory medium and milk: Population and single-cell analysis.

This study evaluated the synergetic inhibitory effects of ultrasound and nisin/carvacrol on spore germination, outgrowth, and subsequent growth of vegetative cell of Bacillus subtilis in laboratory medium and milk. Ultrasound pretreatment (3.33 W/mL, 15 min) and nisin/carvacrol (0.01%, 0.02%) synergistically inhibited spore germination, outgrowth, and vegetative growth of spores in laboratory medium. Whereas no such inhibitory effect was observed in milk even with a 10-fold increase in the concentration (1%) of nisin. Flow cytometry analysis showed that the germination capacities of ultrasound pretreated spores combined with nisin/carvacrol (67.3% and 30.5%, respectively) was lower than that of the untreated spores (95.1%). These results quantitatively revealed the inhibitory effect of the combined treatments which were confirmed by phase-bright spore observations at single cell level. In general, the current work identified the combined ultrasound-carvacrol treatment as an effective strategy to control spores and vegetative cells of B. subtilis in the laboratory medium and milk during abusive storage.

Thermosonication damages the inner membrane of Bacillus subtilis spores and impels their inactivation.

The extreme resistance of bacterial spores to most killing agents makes them a major concern to the food industry and consumers. This gave rise an increasing interest in developing new strategies to inactivate spores and understand the mechanisms of inactivation by various agents. In this study, ultrasound combined with heat (thermosonication, TS) was used to inactivate the spores of Bacillus subtilis and the factors that influence the resistance to TS were analyzed. The spores of wild-type B. subtilis and isogenic mutants were subjected to heat at 80 °C and ultrasound at 6.67-20 W/mL and 23 °C for 0-40 min. TS treatment has synergistically resulted in spore inactivation, and spores of wild-type B. subtilis and isogenic mutants showed different resistance to TS treatment, which was in the following order: Strains 533 (wild-type) ≈ strains PS3518 (gfp) ≈ strains PS2318 (recA-) > strains PS578 (α-ß-), and spores of strains PS3328 (cotE-) were also more susceptible than those of wild-type strains. The inactivated spores lost some proteins in the spore core but initiated germination normally. The germinated inactivated spores did not swell and their plasma membrane permeability was equally altered. It was concluded that the damage to spores' inner membrane (IM) proteins or the IM itself has led to the leakage of intracellular substances and the death of a spore by TS treatment. Our results could support the development and optimization of TS treatment, which has great significance for its further utilization in food industry.

Formation of soy protein isolate (SPI)-citrus pectin (CP) electrostatic complexes under a high-intensity ultrasonic field: Linking the enhanced emulsifying properties to physicochemical and structural properties.

In this study, a high-intensity ultrasonic field was applied to the electrostatic interactions between soy protein isolate (SPI) and citrus pectin (CP). The emulsifying properties of SPI-CP soluble complexes formed under different ultrasound powers and durations were investigated and peaked at 630 W for 10 min. Micrographs of emulsions revealed that ultrasound-treated complexes generated a more homogeneous emulsion with significantly reduced and uniformly-distributed droplet sizes. To better understand the mechanism for the improved emulsifying properties, the physicochemical and structural properties of the SPI-CP complexes at pH 3.5 with and without ultrasound treatment were investigated. It was revealed that ultrasound increased the absolute values of the zeta potential and surface hydrophobicity of complexes, but significantly decreased their particle sizes, fluorescence intensity and turbidity. Results indicated that cavitation effects resulted in structural modifications in both biomacromolecules, as well as enhanced the electrostatic interactions between SPI and CP, which in combination contributed to the more desirable emulsifying properties of the complex.

Activation and conformational changes of chitinase induced by ultrasound.

This study investigated the effect of ultrasound on chitinase activity and conformational changes. Results revealed that ultrasound activated chitinase with a maximum enhancement of 19.17% compared with the untreated chitinase. Furthermore, an increase of Vmax and a decrease of Km after sonication were obtained, illustrating that the affinity between chitinase and substrate was intensified. No obvious effect on the tolerance to most metal ions was exhibited whether sonicated or not (p > 0.05). The conformational changes of chitinase were analyzed by circular dichroism (CD), Fourier transform infrared (FTIR), Raman and fluorescence spectroscopy. Results indicated that the activation of chitinase induced by ultrasound was presumably due to the decrease of tryptophan on the chitinase surface and the increase of ß-sheet and random coil in chitinase secondary conformation. In brief, ultrasound is a possible way to activate chitinase to increase its application in food industry.

Synergistic inactivation and mechanism of thermal and ultrasound treatments against Bacillus subtilis spores.

Some Bacillus species are causative agents of food spoilage and a wide array of diseases. Due to their ability to form highly heat-resistant spores, it is of great interest to develop more effective inactivation strategies whereby these spores could be inactivated. Therefore, this work assessed inactivation of thermal and ultrasound treatments against Bacillus subtilis spores. The study further investigated the thermosonication (thermal and ultrasound, TS) -induced inactivation to the spores through a combination of morphology observation and internal factor analyses. The results of TS indicated that the TS combination synergistically inactivated spores by the maximum log reduction of 2.43 ±â€¯0.08 at 80 °C and 20 W/ml and caused severe cell damage. The visual images revealed that the destructive mode of action of TS had multitarget sites, including coat, cortex, and inner membrane. Three distinct sub-populations were detected by Flow cytometry (FCM), and an unknown step with some physical compromise of the spore's inner membrane and partially hydrolyzed cortex involving the three steps model of inactivation was suggested. The combination of DPA (pyridine-2,6 dicarboxylic acid) content and the relative viabilities of the fractions suggested that during the TS treatment DPA release took place largely after spore death. The dead spores that retained DPA germinated relatively normally, but outgrow poorly, indicating that some key enzymes of intermediary metabolism has been damaged by TS treatment. Such understanding of the lethal action of TS may lead to the development of novel strategies involving spore destruction.

Degradation of carboxymethylcellulose using ultrasound and ß-glucanase: Pathways, kinetics and hydrolysates' properties.

In order to provide an efficient way to degrade carboxymethylcellulose (CMC), three pathways were investigated: enzymolysis, combination of ultrasound pretreatment and enzymolysis, and sonoenzymolysis. Effects of these treatments on enzymatic kinetics, degradation kinetics and properties of degraded CMC were investigated. The degradation degree of CMC was increased by 18.90% and 35.73% with ultrasound pretreatment (at an intensity of 24 W/mL for 30 min) and sonoenzymolysis (at an intensity of 9 W/mL for 50 min), compared with that obtained under the traditional enzymolysis. Analysis of kinetics demonstrated that ultrasound, both pretreatment and combined with ß-glucanase, could accelerate CMC degradation. Measurements of rheological properties, molecular weight and structures of CMC hydrolysates revealed that ultrasound broke the glycosidic bond of CMC chains without changing its primary structure. The sonoenzymolysis process was the most efficient method to degrade CMC, with potential to provide a way to obtain CMC with lowest molecular weight or viscosity.

Alkali solution extraction of rice residue protein isolates: Influence of alkali concentration on protein functional, structural properties and lysinoalanine formation.

This study evaluated the nutrient property and safety of the rice residue protein isolates (RRPI) product (extracted by different alkali concentrations) by exploring the protein functional, structural properties and lysinoalanine (LAL) formation. The results showed that with the rising of alkali concentration from 0.03M to 0.15M, the solubility, emulsifying and foaming properties of RRPI increased at first and then descended. When the alkali concentration was greater than 0.03M, the RRPI surface hydrophobicity decreased and the content of thiol and disulfide bond, Lys and Cys significantly reduced. By the analysis of HPLC, the content of LAL rose up from 276.08 to 15,198.07mg/kg and decreased to 1340.98mg/kg crude protein when the alkali concentration increased from 0.03 to 0.09M and until to 0.15M. These results indicated that RRPI alkaline extraction concentration above 0.03M may cause severe nutrient or safety problems of protein.