Evaluation of antioxidant peptides generated from Jiuzao (residue after Baijiu distillation) protein hydrolysates and their effect of enhancing healthy value of Chinese Baijiu.

BACKGROUND: Jiuzao is the residue after Bajiu distillation which is usually used as forage for livestock. However, it is not fully utilized yet considering the content of protein remained. The present study aimed to isolate antioxidant peptides from Jiuzao protein hydrolysates, then add these peptides into Baijiu product to enhance the healthy value of Baiju. Meanwhile environmental pollution caused by massive Jiuzao can be mitigated indirectly. RESULTS: Four peptides Ala-Tyr-Ile(Leu) (AYI(L)) and Asp-Arg-Glu-Ile(Leu) (DREI(L)) were identified from Jiuzao protein hydrolysates, the extraction contents of AYI + AYL and DREI + DREL were 896.10 and 110.51 mg kg-1 Jiuzao, respectively. On the one hand, antioxidant activities of these peptides were investigated. For in vitro antioxidant assays, AYI, AYL and DREI exhibited strong capacities in oxygen radical absorbance capacity (ORAC) assay. Furthermore, three levels of four peptides were assessed by 2,2'-azobis(2-methylpropanimidamidine) (AAPH)-induced HepG2 cells model. The results showed that these peptides exerted a degree of antioxidant activities in cells. Meanwhile, selected peptides concentrations according to cell assays remined at effective doses after in vitro digestion. On the other hand, the influence of these four peptides on the characteristic aroma compounds in Baijiu was studied. Most characteristic aroma compounds releases were increased with the addition of peptides. CONCLUSION: In the study, antioxidant activities of peptides were evaluated, the feasibility of utilizing Jiuzao protein hydrolysates to obtain beneficial peptides was also proved. Healthy effect of Baijiu or other food can be increased by adding these functional substances. The findings might contribute to food application and Baijiu industries. © 2019 Society of Chemical Industry.

Binding of aldehydes to myofibrillar proteins as affected by two-step heat treatments.

BACKGROUND: The present study investigated the effect of two-step heat treatments on the structure of grass carp myofibrillar proteins (MPs) and their binding ability for selected aldehydes (hexanal, heptanal, octanal and nonanal). RESULTS: Within 30 min of the first heating step at 40 °C and 5-10 min of the second heating step at 90 °C, the enhancement of the flavor-binding ability was likely explained by the increases in surface hydrophobicity and total sulfhydryl content due to the unfolding of secondary structures of MPs through exposure of hydrophobic amino acids and sulfhydryl groups. Nevertheless, lengthy heating at 90 °C accelerated the aggregation of unfolded MPs and reduced the hydrophobic bonding sites, thus weakening the hydrophobic interactions and decreasing the resultant binding ability of MPs with aldehydes. CONCLUSION: The binding ability of aldehydes to MPs was found to be strongly influenced by changes in protein structure and surface during the two-step heating process. The results provided insight into improving the flavor characteristics of freshwater fish surimi products. © 2019 Society of Chemical Industry.

Fluorometric microplate-based dimethoate assay using CdSe/ZnS quantum dots coated with a molecularly imprinted polymer.

A microplate-based assay is described for the sensitive and selective fluorometric determination of the pesticide dimethoate. Molecularly imprinted polymer (MIP)-coated CdSe/ZnS quantum dots (QDs) are used as the molecular recognition probe. The MIP-coated QDs were synthesized using one-step reversed-phase microemulsion in the presence of hydrophobic CdSe/ZnS QDs. Copolymerization was performed by using 3-aminopropyltriethoxysilane as the functional monomer, tetraethoxysilane as the cross-linker, and dimethoate as the template. The fluorescence of the coated QDs is quenched by dimethoate when dimethoate becomes rebound on the imprinting sites of the MIP. Under the optimal conditions, fluorescence (best measured at excitation/emisison wavelengths of 380/620 nm) drops linearly in the 5.0-150 µg L-1 dimethoate concentration range, and the limit of detection is 2.1 µg L-1 (at S/N = 3). The assay was utilized for dimethoate determination in spiked real samples. Satisfactory recoveries (89.8%-98.0%) with relatively standard deviations of <4.9% are obtained. The method is rapid, cost-effective, sensitive, and selective. The use of microplate allows for the quantitation of a large number of samples simultaneously. Graphical abstract Schematic representation of sensitive and selective fluorometric microplate-based assay for the high-throughput determination of dimethoate (DM) based on recognition of molecularly imprinted polymer (MIP)-coated CdSe/ZnS quantum dots (QDs). DM exerts a quenching effect on the fluorescence of the QDs.

A fluorescent nanoprobe for 4-ethylguaiacol based on the use of a molecularly imprinted polymer doped with a covalent organic framework grafted onto carbon nanodots.

An efficient and rapid fluorescent nanoprobe is described for the determination of the aroma compound 4-ethylguaiacol (4-EG). A molecularly imprinted polymer was doped with a covalent organic framework (COF) grafted onto carbon nanodots that was prepared by one-pot reverse microemulsion polymerization. Amino groups at the surface of carbon nanodots coordinate to the COFs to produce a strong bond and this warrants thermal and chemical stability of the probe. Remaining free amino groups interact with the phenolic hydroxyl groups of 4-EG through acid-base pairing interactions. The probe, with excitation/emission maxima at 350/440 nm, responds to 4-EG due to the charge transfer to the carbon nanodots. Under optimized conditions, fluorescence drops linearly as the concentrations of 4-EG increase from 0.025 to 1 µg mL-1, with a detection limit of 17 ng mL-1. The probe was applied to the determination of 4-ethylguaiacol in Chinese Baijiu and wine samples after pretreatment by a single dilution step. The recoveries of spiked samples ranged from 78.4to 110.1%. Graphical abstract Schematic presentation of the synthesis of a fluorescent nanoprobe based on molecularly imprinted polymers doped with covalent organic framework grafted onto carbon nanodots. It was used as an efficient and rapid nanoprobe for 4-ethylguaiacol detection with high selectivity and sensitivity.

Interactions between p-Cresol and Ala-Lys-Arg-Ala (AKRA) from Sesame-Flavor-Type Baijiu.

The objective of this study was to investigate the influence of Ala-Lys-Arg-Ala (AKRA), a tetrapeptide newly identified from Chinese Sesame flavor-type Baijiu in our previous work, on its binding ability with aroma compounds. A method using headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC/MS) was employed to determine the corresponding binding ability between different concentrations of AKRA (10 - 1000 µg/L) and aromas in Baijiu. In comparison, the aroma molecules in Baijiu before and after adding AKRA were almost the same; only their volatility was altered, with mostly the esters, alcohols, and phenolic compounds being decreased. Additionally, AKRA slightly changed the whole aroma profile of Baijiu, especially with respect to p-cresol (an off-odor in sesame-flavor-type Baijiu), which was verified by systematic sensory experiments including gas chromatography-olfactometry (GC-O) and odor activity values (OAVs) analysis. The nonvolatile tetrapeptide favored the hydrogen bond interactions with p-cresol, forming the AKRA- p-cresol complex, thus reducing the headspace concentration of phenolic off-flavors as evidenced by HS-SPME-GC/MS, ultraviolet (UV) absorption spectroscopy, and nuclear magnetic resonance spectrum (1H NMR) analysis. Meanwhile, the hydrogen bond interaction was further confirmed with the intermolecular cross-peaks between the carboxyl hydrogen of AKRA and the phenolic hydroxyl of p-cresol by nuclear Overhauser effect spectroscopy (1H-1H NOESY).

Metal-Free Hydrogen Atom Transfer from Water: Expeditious Hydrogenation of N-Heterocycles Mediated by Diboronic Acid.

A hydrogenation of N-heterocycles mediated by diboronic acid with water as the hydrogen atom source is reported. A variety of N-heterocycles can be hydrogenated with medium to excellent yields within 10 min. Complete deuterium incorporation from stoichiometric D2 O onto substrates further exemplifies the H/D atom sources. Mechanism studies reveal that the reduction proceeds with initial 1,2-addition, in which diboronic acid synergistically activates substrates and water via a six-membered ring transition state.

The Occurrence of Propyl Lactate in Chinese Baijius (Chinese Liquors) Detected by Direct Injection Coupled with Gas Chromatography-Mass Spectrometry.

As one of the oldest distillates in the world, flavor compounds of Chinese Baijiu (Chinese liquor) were extremely complex. Propyl lactate was firstly detected by direct injection and gas chromatography-mass spectrometry (GC-MS) in 72 Chinese Baijius. The objectives were to detect the contents of propyl lactate and evaluate its contribution to the aroma of Chinese Baijiu based on odor activity values (OAVs). The levels of propyl lactate in these distillates were determined by internal standard method and selective ion monitoring (SIM), which ranged from 0.050 to 1.900 mg∙L(-1) under investigation. Its detection threshold was determined by Three-Alternative Forced-Choice (3-AFC) and curve fitting (CF), which was 0.740 mg∙L(-1) in 38% ethanol solution. The contribution of propyl lactate on the aroma of these distillate drinks was evaluated by their odor activity values (OAVs), which varied from 0.066 to 4.440. The OAVs of propyl lactate were found to exceed 1 in 13 Chinese Baijius, including 50° Jingzhi Guniang 5 years (4.440), 52° Jingzhi Guniang 10 years (3.024), Jingyanggang (2.568), Xianghe Ronghe Shaofang (2.313), and 1956 Laolang (1.431), which indicated that propyl lactate was one of odor-active components in these Chinese Baijius.

Based on metabolomics, chemometrics, and modern separation omics: Identifying key in-pathway and out-pathway points for pesticide residues during solid-state fermentation of baijiu.

By collecting real samples throughout the entire production process and employing chemometrics, metabolomics, and modern separation omic techniques, it unveiled the patterns of pesticide transfer during solid-state fermentation. The results indicated that 12 types of pesticide residues were prevalent during baijiu production, with organochlorine and carbamate pesticides being the most abundant in raw materials. After fermentation, organochlorine pesticides and pyrethroid pesticides exhibited higher content, while carbamate pesticides dominated in the final product. The pathways for pesticide input and elimination were identified, and the intricate mechanisms underlying these changes were further elucidated. Additionally, key control points were defined to facilitate targeted monitoring. The results indicated that pesticide residue primarily originates from raw materials and Daqu, whereas both solid-state fermentation and distillation processes were effective in reducing pesticide residues. The study offers valuable guidance for establishing pesticide residue standards in the context of baijiu production.

Decoding Molecular Mechanism Underlying Human Olfactory Receptor OR8D1 Activation by Sotolone Enantiomers.

Sotolone, a chiral compound, plays an important role in the food industry. Herein, (R)-/(S)-sotolone were separated to determine their odor characteristics and thresholds in air (R-form: smoky, burned, herb, and green aroma, 0.0514 µg/m3; S-form: sweet, milk, acid, and nutty aroma, 0.0048 µg/m3). OR8D1 responses to (R)-/(S)-sotolone were detected in a HEK293 cell-based luminescence assay. (S)-Sotolone was a more potent agonist than (R)-sotolone (EC50 values of 84.98 ± 1.05 and 167.20 ± 0.25 µmol/L, respectively). Molecular dynamics simulations and molecular mechanics Poisson-Boltzmann surface area analyses confirmed that the combination of (S)-sotolone and OR8D1 was more stable than that of (R)-sotolone. Odorant docking, multiple sequence alignments, site-directed mutagenesis, and functional studies with recombinant odorant receptors (ORs) in a cell-based luminescence assay identified 11 amino-acid residues that influence the enantioselectivity of OR8D1 toward sotolone significantly and that N2065.46 was indispensable to the activation of OR8D1 by (S)-sotolone.

Nanotechnology-based optical sensors for Baijiu quality and safety control.

Baijiu quality and safety have received considerable attention owing to the gradual increase in its consumption. However, owing to the unique and complex process of Baijiu production, issues leading to quality and safety concerns may occur during the manufacturing process. Therefore, establishing appropriate analytical methods is necessary for Baijiu quality assurance and process control. Nanomaterial (NM)-based optical sensing techniques have garnered widespread interest because of their unique advantages. However, comprehensive studies on nano-optical sensing technology for quality and safety control of Baijiu are lacking. In this review, we systematically summarize NM-based optical sensor applications for the accurate detection and quantification of analytes closely related to Baijiu quality and safety. Furthermore, we evaluate the sensing mechanisms for each application. Finally, we discuss the challenges nanotechnology poses for Baijiu analysis and future trends. Overall, nanotechnological approaches provide a potentially useful alternative for simplifying Baijiu analysis and improving final product quality and safety.

meso-α,α-5,15-Bis(o-nicotinamido-phen-yl)-10,20-diphen-ylporphyrin n-hexane monosolvate.

The structure of the title solvated porphyrin, C56H38N8O2·C6H14, is reported. Two porphyrin mol-ecules, one ordered and one disordered n-hexane solvate mol-ecules are present in its asymmetric unit. The porphyrin macrocycle shows a characteristic saddle-shaped distortion, and the maximum deviation from the mean plane for non-hydrogen atoms is 0.48 Å. N-H⋯N, N-H⋯O, and C-H⋯O hydrogen bonds, as well as π-π inter-actions, are observed in the crystal structure.

Investigation on the key factors associated with flavor quality in northern strong aroma type of Baijiu by flavor matrix.

The strong aroma type of Baijiu is popular with its balanced flavor. However, the flavor characteristics of strong aroma type of Baijiu in different regions have significant diversities. Among them, the key aroma compounds in northern strong aroma type of Baijiu and the factors affecting the aroma profile and flavor quality are still unclear. In this study, a total of 78 aroma compounds were identified by gas chromatography-olfactometry-mass spectrometry (GC-O-MS). The quantitative analysis was completed and ethyl hexanoate (1003.24-4506.04 mg/L) was the highest. Aroma profile was successfully simulated by reconstitution, and omission test was applied to investigate the effects of the important aroma compounds on the aroma profile. Additionally, the relationship between expression of aroma compounds and sensory characteristics of samples was illustrated by flavor matrix. The overall aroma profile of northern strong aroma type of Baijiu was formed by the key aroma compounds with different aroma expression.

Reducing the background interference of liquid-liquid extraction method during Baijiu aroma analysis.

Liquid-liquid extraction coupled with gas chromatography-mass spectrometry (LLE-GC/MS) is a useful methodology for Baijiu aroma analysis. However, the background problems caused by LLE have rarely been investigated. In the present study, synthetic Baijiu was analyzed by traditional LLE-GC/MS to assess background problems, and a series of ghost peaks of some compounds were detected within the retention time of 47.0-60.0 min, including 2,4-di-tert-butylphenol (2,4-DTBP), palmitic acid and oleanitrile. 2,4-DTBP, which has a phenolic odor, is an important component in both strong and light-flavor Baijiu. Single-factor experiments confirmed that 2,4-DTBP is not a constituent of Baijiu but migrates from sodium sulphate and sodium chloride, and, high-temperature baking was an effective approach to eliminate these compounds. A combined strategy using standard cleaning coupled with high-temperature baking was proposed to reduce the background problems during LLE. Furthermore, more trace compounds could be identified through this process in the future.

Succession patterns of aroma components during brewing process of broomcorn millet (Panicum miliaceum L.) Huangjiu.

The flavor of Huangjiu is closely related to its brewing technology. Patterns of aroma component succession during the process of brewing broomcorn millet Huangjiu were investigated by solvent-assisted flavor evaporation combined with gas chromatography-mass spectrometry and chemometrics. During fermentation, esters, alcohols, acids, ketones, acetals, sulfur compounds, furans, and lactones were formed mostly in the chief fermentation stage; nitrogenous and phenolic compounds increased in the primary fermentation stage and then decreased; aldehydes decreased after fermentation started; and terpenes decreased after five days. During aging, acids, alcohols, ketones, lactones, phenols, and nitrogenous and sulfur compounds first decreased and then increased; and esters, acetals, aldehydes, and furans always increased, while terpenes decreased continuously. Key odorants, including acetic acid, 3-methylbutanoic acid, 1,1-diethoxyethane, and 3-methylbutanal, were produced in large quantities in the primary fermentation stage; ethyl lactate, ß-phenylethanol, and 2/3-methyl-1-butanol were generated in large quantities in the chief fermentation stage; and sotolon and methional were generated in the aging stage. This study is of great significance for the quality control of Huangjiu production.

Insights into a new alternative method with graphene oxide/polyacrylamide/Fe3O4 nanocomposite for the extraction of six odor-active esters from Strong-aroma types of Baijiu.

Liquid-liquid extraction (LLE) is the most commonly utilized technique for the extraction of odor-active esters (OAEs) in strong-aroma types of Baijiu (SAB). However, since the contents of different OAEs in SAB vary widely, it is still a puzzle to ensure that all OAEs to be thoroughly extracted by LLE without the problem of saturated adsorption. Herein, a novel approach of magnetic solid phase extraction (MSPE), based on the magnetic graphene oxide nanocomposite modified with polyacrylamide (GO/PAM/Fe3O4), was employed for the efficient extraction of six OAEs from SAB. Compared with LLE, GO/PAM/Fe3O4 exhibited highly selective recognition properties and larger adsorption capacities for OAEs (ranging from 13.68 to 39.06 mg/g), resulting in better extraction performances for OAEs. Coupled with GC-MS, six OAEs in real SAB were successfully determined, with recoveries ranged from 70.1 âˆ¼ 90.0% and LODs at 0.08 âˆ¼ 1.35 µg/L. Overall, the MSPE-GC/MS is a promising alternative for accurate determination of OAEs in SAB.

Quick classification of strong-aroma types of base Baijiu using potentiometric and voltammetric electronic tongue combined with chemometric techniques.

Nowadays, the classification of strong-aroma types of base Baijiu (base SAB) is mainly achieved by human sensory evaluation. However, prolonged tasting brings difficulties for sommeliers in guaranteeing the consistency of results, and may even cause health problems. Herein, an electronic tongue (E-Tongue) combined with a gas chromatography-mass spectrometry (GC-MS) method was successfully developed to grade high-alcoholic base SAB. The E-tongue was capable of identifying base SAB samples into four grades by a discriminant function analysis (DFA) model based on human sensory evaluation results. More importantly, it could effectively and rapidly predict the quality grade of unknown base SAB with an average accuracy up to 95%. The differences of chemical components between base SAB samples were studied by the GC-MS analysis and 52 aroma compounds were identified. The qualitative and quantitative results showed that with the increase of base SAB grade, the varieties and contents of aroma compounds increased. Overall, the comprehensive analysis of E-tongue data and GC-MS results could be in good agreement with human sensory evaluation results, which also proved that the newly developed method has a potential to be a useful alternative to the overall quality grading of base Baijiu.

Characterization and comparative study of the key odorants in Caoyuanwang mild-flavor style Baijiu using gas chromatography-olfactometry and sensory approaches.

Caoyuanwang Baijiu (CYW), a mild-flavor style Baijiu (MSB), is popular in northern China. However, there is a lack of studies reporting its aroma-active components. The aroma compounds of five CYW samples were analyzed using gas chromatography-olfactory-mass spectrometry coupled with aroma extraction dilution analysis. Fifty-five aroma-active compounds were identified in CYW, of which 27 had odor activity values ≥ 1. Reconstituted models successfully simulated the aroma profiles of CYW. The omission tests elucidated that ß-damascenone, dimethyl trisulfide, ethyl pentanoate, butanoic acid, ethyl acetate, 3-methylbutanal, ethyl lactate, hexanoic acid, γ-nonalactone, 3-hydroxy-2-butanone, ethyl butanoate, 1-propanol, 4-(ethoxymethyl)-2-methoxy-phenol, and vanillin were key odorants in CYW. The addition test confirmed the significant influence of dimethyl trisulfide on Chen-aroma note. Nine key odorants were identified as the differential quality-markers, and 85.71% key odorants were predicted using the partial least square regression (PLSR) analysis, indicating the applicability of PLSR in selecting the target compounds for omission tests.

An efficient phthalate ester-degrading Bacillus subtilis: Degradation kinetics, metabolic pathway, and catalytic mechanism of the key enzyme.

Phthalate ester pollution in the environment and food chain is frequently reported. Microbial treatment is a green and efficient method for solving this problem. The isolation and systematic investigation of microorganisms generally recognized as safe (GRAS) will provide useful resources. A GRAS Bacillus subtilis strain, BJQ0005, was isolated from Baijiu fermentation starter and efficiently degraded phthalate esters (PAEs). The half-lives for di-isobutyl phthalate, di-butyl phthalate and di-(2-ethylhexyl) phthalate were 3.93, 4.28, and 25.49 h, respectively, from the initial amount of 10 mg per 10 mL reaction mixture, which are records using wild-type strains. Genome sequencing and metabolic intermediate analysis generated the whole metabolic pathway. Eighteen enzymes from the α/ß hydrolase family were expressed. Enzymes GTW28_09400 and GTW28_13725 were capable of single ester bond hydrolysis of PAEs, while GTW28_17760 hydrolyzed di-ester bonds of PAEs. Using molecular docking, a possible mechanism affecting enzymatic ester bond hydrolysis of mono-butyl phthalate was proposed of GTW28_17760. The carboxyl group generated by the first hydrolysis step interacted with histidine in the catalytic active center, which negatively affected enzymatic hydrolysis. Isolation and systematic investigation of the PAE degradation characteristics of B. subtilis will promote the green and safe treatment of PAEs in the environment and food industry.

Discovery and development of a novel short-chain fatty acid ester synthetic biocatalyst under aqueous phase from Monascus purpureus isolated from Baijiu.

Short-chain fatty acid esters are important flavor chemicals in Chinese traditional fermented Baijiu. Monascus purpureus was recognized as an important microorganism contributing to ester synthesis. However, the molecular basis for ester synthesis was still lacking. The present work combined genome sequencing, transcriptome sequencing, gene library construction, and enzyme engineering to discover a novel catalyst from M. purpureus (isolated from Baijiu fermentation starter). Enzyme LIP05, belonging to the α/ß hydrolase family, was identified to synthesize short-chain fatty acid esters under aqueous phase. After deleting the lid domain of LIP05, the synthesis of ethyl pentanoate, ethyl hexanoate, ethyl octanoate, or ethyl decanoate was achieved. Ethyl octanoate with the highest conversion ratio of 93.7% was obtained with the assistance of ultrasound. The study reveals the molecular basis for synthesizing short-chain fatty acid esters by M. purpureus and will promote the application of the species or the enzyme in food industry.

Characterization of Key Aroma Compounds in Tartary Buckwheat (Fagopyrum tataricum Gaertn.) by Means of Sensory-Directed Flavor Analysis.

The key odorants of tartary buckwheat (TB) were researched by a sensory-directed flavor analysis approach for the first time. After the volatiles of TB were isolated by solvent-assisted flavor evaporation (SAFE), 49 aroma-active components with flavor dilution (FD) factors in the range of 1-2187 were identified using gas chromatography-olfactometry-mass spectrometry (GC-O-MS) combined with aroma extract dilution analysis (AEDA). Geranylacetone, phenethyl alcohol, and ß-damascone showed the highest FD factors of 2187. All 49 odorants were further quantitated by the internal standard curve method, and their odor activity values (OAVs) were obtained. The overall aroma of TB was successfully simulated (similarity > 98.16%) by mixing 16 odorants (OAV ≥ 1) with their natural concentrations. The omission tests revealed that geosmin, α-isomethylionone, α-methylionone, ß-ionone, linalool, ß-damascone, geranylacetone, guaiacol, ethyl hexanoate, geraniol, vanillin, tetrahydrolinalool, and 2,5-dimethyl-4-hydroxy-3-(2H)-furanone were the key odorants of TB. Chiral analysis showed that tetrahydrolinalool and linalool existed as racemics in the commercial TB. The relative content of R-enantiomers of α-isomethylionone and α-methylionone was slightly higher than that of their S-enantiomers. The odor thresholds of R- and S-enantiomer of tetrahydrolinalool were first detected as 0.029 and 3.8 µg/L in air, respectively.