Structural characterization and angiotensin-converting enzyme (ACE) inhibitory mechanism of Stropharia rugosoannulata mushroom peptides prepared by ultrasound

To reveal the structural characteristics and angiotensin-converting enzyme (ACE) inhibition mechanism of Stropharia rugosoannulata mushroom peptides prepared by multifrequency ultrasound, the peptide distribution, amino acid sequence composition characteristics, formation pathway, and ACE inhibition mechanism of S. rugosoannulata mushroom peptides were studied. It was found that the peptides in S. rugosoannulata mushroom samples treated by multifrequency ultrasound (probe ultrasound and bath ultrasound mode) were mainly octapeptides, nonapeptides, and decapeptides. Hydrophobic amino acids were the primary amino acids in the peptides prepared by ultrasound, and the amino acid dissociation of the peptide bonds at the C-terminal under the action of ultrasound was performed mainly to produce hydrophobic amino acids. Pro and Val (PV), Arg and Pro (RP), Pro and Leu (PL), and Asp (D) combined with hydrophobic amino acids were the characteristic amino acid sequence basis of the active peptides of the S. rugosoannulata mushroom. The docking results of active peptides and ACE showed that hydrogen bond interaction remained the primary mode of interaction between ACE and peptides prepared by ultrasound. The peptides can bind to the amino acid residues in the ACE active pocket, zinc ions, or key amino acids in the domain, and this results in inhibition of ACE activity. Cation–pi interactions also played an important role in the binding of mushroom peptides to ACE. This study explains the structural characteristics and ACE inhibition mechanism used by S. rugosoannulata mushroom peptides prepared by ultrasound, and it will provide a reference for the development and application of S. rugosoannulata mushroom peptides.     

Valine 738 and lysine 735 in the fifth transmembrane domain of rTas1r3 mediate insensitivity towards lactisole of the rat sweet taste receptor

Background  

The sweet taste inhibitor lactisole acts on the human sweet taste receptor heteromer TAS1R2-TAS1R3 but not on its rodent counterpart. Recently, it was shown that the lactisole sensitivity of the human sweet taste receptor involves the part of TAS1R3 encompassing the seven transmembrane regions but not the huge N-terminal domain. Using mutational analysis we investigated which amino acid residues distinguish lactisole insensitive rat from sensitive human T1R3 receptors.     

Expression of Galpha14 in sweet-transducing taste cells of the posterior tongue

Background  

"Type II"/Receptor cells express G protein-coupled receptors (GPCRs) for sweet, umami (T1Rs and mGluRs) or bitter (T2Rs), as well as the proteins for downstream signalling cascades. Transduction downstream of T1Rs and T2Rs relies on G-protein and PLCβ2-mediated release of stored Ca²⁺. Whereas Gαgus (gustducin) couples to the T2R (bitter) receptors, which Gα-subunit couples to the sweet (T1R2 + T1R3) receptor is presently not known. We utilized RT-PCR, immunocytochemistry and single-cell gene expression profiling to examine the expression of the Gαq family (q, 11, 14) in mouse taste buds.     

Sonolytic hydrolysis of peptides in aqueous solution upon addition of catechol

The sonolytic hydrolysis of peptides with addition of phenolic reagents to aqueous solutions is described. Sonolysis of an aqueous solution of peptides to which catechol (o-dihydroxybenzene) had been added resulted in hydrolytic products reflecting the amino acid sequence without any side reactions, while sonolysis without any additives resulted in oxidation analytes and degradation products caused by side reactions. Although the use of additives such as resorcinol (m-dihydroxybenzene), hydroquinone (p-dihydroxybenzene) and phenol was also effective in producing sequence related products, several degradation products were produced by side reactions. A characteristic of the sonolysis of peptides is that the N-terminal side of proline, Xxx-Pro, is more susceptible than other amino acid residues to the process. This characteristic of sonolysis is superior to that of acid hydrolysis in which cleavage at the C-terminal side of proline, Pro-Xxx is difficult, and where dehydration products result due to side reactions.     

The effect of high hydrostatic pressure on taste substance distribution in fresh green tea leaves

ABSTRACT

High hydrostatic pressure (HHP) can be an alternative method to steaming to inhibit enzymatic fermentation in green tea making process. However, the effect of HHP treatment on green tea taste is not clear. Thus, this study aimed to determine the effect of HHP on substances associated with green tea taste. Fresh green tea leaves were immediately treated with HHP at 300, 500, or 700?MPa for 10, 30, or 60 min at 25°C. The concentration of free amino acids, catechins, and caffeine in HHP-treated samples was quantified by LC-MS. The taste intensity of the samples was detected by taste sensors. HHP resulted in a high accumulation of free amino acids in green tea leaves, which was likely due to proteolysis. In particular, theanine synthesis may have been promoted by an increase in the concentration of substrates during HHP. Compared to steaming, HHP enhanced umami richness, and inhibited bitterness and astringency.     

Effects of ultrasound treatment on muscle structure,volatile compounds,and small molecule metabolites of salted Culter alburnus fish

This study investigated the effects of ultrasound treatment on the quality of salted Culter alburnus fish. The results showed that with the increasing ultrasound power, the structural degradation of muscle fibers was intensified, and the conformation of myofibrillar protein was significantly changed. The high-power ultrasound treatment group (300 W) had relatively higher thiobarbiturate reactive substance content (0.37 mg malondialdehyde eq/kg) and peroxidation value (0.63 mmol/kg). A total of 66 volatile compounds were identified with obvious differences among groups. The 200 W ultrasound group exhibited fewer fishy substances (Hexanal, 1-Pentene-3-ol, and 1-Octane-3-ol). Compared with control group, ultrasound groups (200, 300 W) contained more umami taste-related amino peptides such as γ-Glu-Met, γ-Glu-Ala, and Asn-pro. In the ultrasound treatment group, L-isoleucine and L-methionine, which may be used as flavor precursors, were significantly down-regulated, while carbohydrates and its metabolites were up-regulated. Amino acid, carbohydrate, and FA (fatty acyls) metabolism products in salted fish were enriched by ultrasound treatment, and those products might ultimately be related to the taste and flavor of salted fish.     

Influence of ultrasound-assisted ionic liquid pretreatments on the functional properties of soy protein hydrolysates

In this work, the effect of dual-frequency ultrasound-assisted ionic liquids (ILs) pretreatment on the functional properties of soy protein isolate (SPI) hydrolysates was investigated. The degree of hydrolysis (DH) of SPI pretreated by ultrasound and [BMIM][PF₆] increased by 12.53% as compared to control (P < 0.05). More peptides with low molecular weight were obtained, providing support for the changes in DH. The trichloroacetic acid-nitrogen soluble index presented an increase, suggesting a better protein hydrolysate property. The increase in the calcium-binding activity showed the ultrasound-assisted ILs pretreatment could potentially improve bone health. The foaming capacity and stability of SPI hydrolysates pretreated by ultrasound-assisted [BMIM][PF₆] always increased remarkably as compared to ultrasound-assisted [BDMIM][Cl] pretreatment. However, the synergistic effect of ultrasound-assisted [BMIM][PF₆] on the emulsifying activity and antioxidant activities (DPPH and hydroxyl radical scavenging activity) was not as ideal as ultrasound-assisted [BDMIM][Cl] pretreatment, which may be affected by the structure of peptide. In conclusion, these results indicated the combination of dual-frequency ultrasound and ionic liquids would be a promising method to improve the functional properties of SPI hydrolysates and broaden the application scope of compound modification in proteolysis industry.     

Evidence for a role of glutamate as an efferent transmitter in taste buds

Background  

Glutamate has been proposed as a transmitter in the peripheral taste system in addition to its well-documented role as an umami taste stimulus. Evidence for a role as a transmitter includes the presence of ionotropic glutamate receptors in nerve fibers and taste cells, as well as the expression of the glutamate transporter GLAST in Type I taste cells. However, the source and targets of glutamate in lingual tissue are unclear. In the present study, we used molecular, physiological and immunohistochemical methods to investigate the origin of glutamate as well as the targeted receptors in taste buds.     

Impact of different ultrasound-assisted processes for preparation of collagen hydrolysates from Asian bullfrog skin on characteristics and antioxidative properties

This study focused on impact of ultrasound-assisted process (UAP) at pre-treatment (UP) and simultaneous treatment (US) during papain hydrolysis for preparing collagen hydrolysate (CH) from Asian bullfrog skin. Ultrasonication times were varied (10, 20, 30 min), and CH prepared using papain hydrolysis without UAP was used as control. Different UAPs provided CH with various hydroxyproline contents, α-amino group contents, surface hydrophobicities, and antioxidative activities. UP at 20 min (UP-20) and US at 30 min (US-30) provided highly antioxidative CHs, which were selected for further studies on their Oxygen reactive absorbance capacity (ORAC) and molecular characteristics. CHs from UP-20 and US-30 had higher ORAC than that of control group (p ≤ 0.05). Slight difference in amino acid composition was detected between samples. Based on these results, molecular characteristic styles, molecular weight profile, antioxidative peptide content, and secondary structure of each sample were obtained. These results indicate that UP and US used varied enzymatic hydrolysis patterns and modified molecular conformation of CH, resulting in enhanced antioxidative activity. Therefore, different UAPs as UP and US could be effectively used in preparation of CH using papain hydrolysis from Asian bullfrog skin, which could improve production process efficiency by enhancing their bioactivity, particularly antioxidative activity.     

Conversion of waste meat to resources by enzymatic reaction under high pressure carbon dioxide conditions

ABSTRACT

Every year, a tremendous amount of meat is wasted, which may have adverse effects on the environment. Therefore, waste meat should be recycled. Building a recycling system for waste meat to reduce financial loss and careful treatment of life-based resources is essential. As meat contains abundant amino acids, it can be used as an immense source of amino acids for other industries following enzymatic hydrolysis. However, during the enzymatic hydrolysis process, the waste meat can easily get contaminated by microbes and therefore needs high temperature. In the present study, we applied high pressure carbon dioxide (HPCD) technology to prevent the contamination of meat by microbes during enzymatic hydrolysis process. According to our research, HPCD has a great potential to be adopted as a novel decontamination technology in the industrial enzymatic hydrolysis process at relatively lower temperature.     

Voltage-gated sodium channels in taste bud cells

Background  

Taste bud cells transmit information regarding the contents of food from taste receptors embedded in apical microvilli to gustatory nerve fibers innervating basolateral membranes. In particular, taste cells depolarize, activate voltage-gated sodium channels, and fire action potentials in response to tastants. Initial cell depolarization is attributable to sodium influx through TRPM5 in sweet, bitter, and umami cells and an undetermined cation influx through an ion channel in sour cells expressing PKD2L1, a candidate sour taste receptor. The molecular identity of the voltage-gated sodium channels that sense depolarizing signals and subsequently initiate action potentials coding taste information to gustatory nerve fibers is unknown.     

Immunocytochemical evidence for co-expression of Type III IP3 receptor with signaling components of bitter taste transduction

Background  

Taste receptor cells are responsible for transducing chemical stimuli into electrical signals that lead to the sense of taste. An important second messenger in taste transduction is IP₃, which is involved in both bitter and sweet transduction pathways. Several components of the bitter transduction pathway have been identified, including the T2R/TRB taste receptors, phospholipase C β2, and the G protein subunits α-gustducin, β3, and γ13. However, the identity of the IP₃ receptor subtype in this pathway is not known. In the present study we used immunocytochemistry on rodent taste tissue to identify the IP₃ receptors expressed in taste cells and to examine taste bud expression patterns for IP₃R3.     

复合酶水解云南白鱼制备生物活性肽类物质

研究了复合蛋白酶对云南土著白鱼水解的工艺条件。考察了酶用量、温度、pH值、反应时间、底物浓度、复合酶比例等因素对白鱼水解的影响。确定了最佳酶解条件为:复合酶用量(木瓜蛋白酶+风味蛋白酶1∶1)16000U/g,底物浓度30%,温度55℃,pH值7.5,时间4h。在此条件下,水解度达34.12%,氮回收率90.05%。该结果为云南白鱼生产功能食品提供科学依据。     

Immunocytochemical Analysis of P2X2 in Rat Circumvallate Taste Buds

ABSTRACT: BACKGROUND: Our laboratory has shown that classical synapses and synaptic proteins are associated with Type III cells. Yet it is generally accepted that Type II cells transduce bitter, sweet and umami stimuli. No classical synapses, however, have been found associated with Type II cells. Recent studies indicate that ionotropic purinergic receptors P2X2/P2X3 are present in rodent taste buds. Taste nerve processes express the ionotropic purinergic receptors (P2X2/P2X3). P2X2/P2X3Dbl/ mice are not responsive to sweet, umami and bitter stimuli, and it has been proposed that ATP acts as a neurotransmitter in taste buds. The goal of the present study is to learn more about the nature of purinergic contacts in rat circumvallate taste buds by examining immunoreactivity to antisera directed against the purinergic receptor P2X2. RESULTS: P2X2-like immunoreactivity is present in intragemmal nerve processes in rat circumvallate taste buds. Intense immunoreactivity can also be seen in the subgemmal nerve plexuses located below the basal lamina. The P2X2 immunoreactive nerve processes also display syntaxin-1-LIR. The immunoreactive nerves are in close contact with the IP3R3-LIR Type II cells and syntaxin-1-LIR and/or 5-HT-LIR Type III cells. Taste cell synapses are observed only from Type III taste cells onto P2X2-LIR nerve processes. Unusually large, "atypical" mitochondria in the Type II taste cells are found only at the close appositions with P2X2-LIR nerve processes. P2X2 immunogold particles are concentrated at the membranes of nerve processes at the close appositions with taste cells. CONCLUSIONS: Based on our fluorescence and immunoelectron microscopical studies we believe that both perigemmal and most all intragemmal nerve processes display P2X2-LIR. Moreover, colloidal gold immunoelectron microscopy indicates that P2X2-LIR in nerve processes is concentrated at sites of close apposition with Type II cells. This supports the hypothesis that ATP may be a key neurotransmitter in taste transduction and that Type II cells release ATP, activating P2X2 receptors in nerve processes.     

Ultrasound microwave-assisted enzymatic production and characterisation of antioxidant peptides from sweet potato protein

Herein, we investigated the effects of ultrasound microwave (UM)-assisted hydrolysis using Alcalase (ALC), Flavourzyme (FLA), and their combination (ALC + FLA), on the production of sweet potato protein hydrolysates (SPPH). UM-assisted enzymatic hydrolysis significantly increased the degree of hydrolysis of SPPH compared with untreated (UN) samples. Fractions with differences in molecular weight (MW) of >10, 3–10, and < 3 kDa in SPPH from UM-assisted ALC, FLA, and ALC + FLA hydrolysis displayed higher antioxidant activities than those from UN samples. MW < 3 kDa fractions of SPPH from UM-assisted ALC and ALC + FLA hydrolysis treatments presented much stronger Fe²⁺-chelating activity (98.48% and 98.59%), ·OH scavenging activity (67.11% and 60.06%), and higher ORAC values (110.32 and 106.32 µg TE/mL), from which diverse peptides with potential antioxidant activities were obtained by semi-preparative HPLC and LC-MS/MS. All identified peptide sequences exhibited at least three potential antioxidant amino acids. Additionally, changes in peptide conformational structure and antioxidant amino acid composition were revealed by structure–activity relationship analysis. Thus, ultrasound microwave treatment has great potential in antioxidant peptides production.     

Effects of ultrasound-assisted heating on aroma profile,peptide structure,peptide molecular weight,antioxidant activities and sensory characteristics of natural fish flavouring

This study deals with the production of natural fish flavouring using ultrasound-assisted heating process. The effect of ultrasound pretreatment at different amplitudes (0, 15, 30, and 45%) on the Maillard reaction rate, antioxidant activities, flavour profile, and sensory characteristics of fish flavouring was investigated. Results showed that sonication markedly accelerated the Maillard reaction (MR) rate, as evidenced by the modification of peptide structure, a decrease in pH value, free amino acid content coupled with a rise in browning intensity. Also, ultrasound pretreatment significantly enhanced the antioxidant activities of fish flavouring (p ≤ 0.05). Moreover, sonication increased the type and content of aroma compounds significantly. Sensory analysis revealed that ultrasound pretreatment increased the fish-like and toasty aroma as well as umami and mouthfulness attributes coupled with the reduction of the bitter taste of the fish flavouring. This result was consistent with the GC–MS, electronic nose, cluster, and Partial Least Squares Regression (PLSR) analyses, which clearly showed that ultrasound pretreatment enhanced the fish-like aroma, which was associated with the increase in aldehydes, ketones, alcohols, thiophenes, pyrazine, and furans contents. Thus, it could be concluded that ultrasound pretreatment, coupled with the thermal process, could be a promising process for the production of natural fish flavouring with higher antioxidant activities.     

Effects of ultrasound-assisted thawing on lamb meat quality and oxidative stability during refrigerated storage using non-targeted metabolomics

The aim of this study was to evaluate the changes of ultrasound-assisted thawing on lamb meat quality and differential metabolite profiles during refrigerated storage. Compared with flow water thawing (FW), pH, a*, C*, and sulfhydryl content of lamb were significantly increased, while L*, drip loss and cooking loss were significantly decreased after ultrasound-assisted thawing (UT). On day 1 (UT1 and FW1) and day 7 (UT7 and FW7) in the UT and FW groups, principal component analysis explained 42.22% and 39.25% of the total variance. In this study, 44 (UT1 and FW1) and 47 (UT7 and FW7) differentially expressed metabolites were identified, including amino acids, carbohydrates and their conjugates, nucleic acids, carbonyl compounds and others. The results of this study provide data to clarify the differences between UT and FW, and lay a foundation for the application of ultrasound-assisted thawing in the meat industry.     

Improved sonolytic hydrolysis of peptides in aqueous solution with addition of 1,4-benzenedithiol

Described here is the sonolytic hydrolysis of peptides achieved by treatment of aqueous solution to which the radical scavenger 1,4-benzenedithiol (1,4-BDT), which has hydrogen donating ability, has been added. Mass spectrometric analysis of the products of sonolytic hydrolysis gave information about amino acid sequence of the peptides without any byproducts. The additive 1,4-BDT improves the sonolytic hydrolysis of peptides in terms of the rate of hydrolysis reaction and the amount of additive required when compared to catechol, a previously reported additive. The sonolytic hydrolysis of peptides differs from both acid hydrolysis and hydrogen atom-induced dissociation named matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD), in characteristics. We propose a mechanistic reaction for the sonolytic hydrolysis of peptides, based on the mechanisms of both acid hydrolysis and MALDI-ISD processes. The sonolytic hydrolysis of peptides upon addition of hydrogen donating radical scavengers can be rationalized via the attachment of a hydrogen atom to the carbonyl oxygen with subsequent hydrolysis.     

Amiloride-sensitive channels in type I fungiform taste cells in mouse

Background  

Taste buds are the sensory organs of taste perception. Three types of taste cells have been described. Type I cells have voltage-gated outward currents, but lack voltage-gated inward currents. These cells have been presumed to play only a support role in the taste bud. Type II cells have voltage-gated Na⁺ and K⁺ current, and the receptors and transduction machinery for bitter, sweet, and umami taste stimuli. Type III cells have voltage-gated Na⁺, K⁺, and Ca²⁺ currents, and make prominent synapses with afferent nerve fibers. Na⁺ salt transduction in part involves amiloride-sensitive epithelial sodium channels (ENaCs). In rodents, these channels are located in taste cells of fungiform papillae on the anterior part of the tongue innervated by the chorda tympani nerve. However, the taste cell type that expresses ENaCs is not known. This study used whole cell recordings of single fungiform taste cells of transgenic mice expressing GFP in Type II taste cells to identify the taste cells responding to amiloride. We also used immunocytochemistry to further define and compare cell types in fungiform and circumvallate taste buds of these mice.     

Investigation on ultrasound-assisted extraction of salvianolic acid B from Salvia miltiorrhiza root

It is reported that salvianolic acid B, a bioactive phenolic compound contained in the root of Salvia miltiorrhiza, exhibits a much stronger activity in free radical scavenging and antioxidance than those of vitamin E. When a conventional refluxing method is adopted to extract salvianolic acid B from the root, in which the materials are subjected to higher temperature and longer time, the yield of this phenolic compound is lower due to the possibility of its hydrolysis to tanshinol. However, a higher extraction yield can be achieved over a shorter time period and lower temperature when an ultrasound-assisted extraction method is used. This paper investigated the parameters influencing the extraction of salvianolic acid B. Factors such as extraction time, frequency of the ultrasound, the ratio of solvent to material, and types of extraction solvent were examined. A comparison was also conducted between conventional refluxing and ultrasound-assisted extraction. Results showed that the optimal parameters to extract salvianolic acid B from the root of S. miltiorrhiza were as follows: ultrasonic frequency: 45 Hz; solvent: 60% aqueous ethanol; extraction temperature: 30 °C; extraction time duration: 25 min.; ratio of solvent to material: 20:1 (v/w, ml/g). Under these conditions, the yield of salvianolic acid B was 5.17 mg/g (33.93 mg/g) higher than those with conventional refluxing method (28.76 mg/g), indicating that the efficiency and the yield of ultrasound-assisted extraction method are higher than reflux method, and the hydrolysis of salvianolic acid B to tanshinol is effectively avoided.