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1.
Food Microbiol ; 122: 104563, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38839237

RESUMO

Thermosonication (UT) prestress treatments combining with varied fermentation patterns has been revealed as an effective method to regulate post-acidification as exerted by Lactobacillus delbrueckii subsp. bulgaricus (L. delbrueckii), but sono-biochemical controlling mechanisms remain elusive. This study employed physiological and transcriptomic analysis to explore the response mechanism of L. delbrueckii to UT-induced microstress (600 W, 33 kHz, 10 min). UT stress-induced inhibition of acidification of L. delbrueckii during (post)-fermentation was first confirmed, relying on the UT process parameters such as stress exposure duration and UT power. The significantly enhanced membrane permeability in cells treated by 600 W for 10 min than the microbes stressed by 420 W for 20 min suggested the higher dependence of UT-derived stresses on the treatment durations, relative to the ultrasonic powers. In addition, ultrasonication treatment-induced changes in cell membrane integrity enhanced and/or disrupted permeability of L. delbrueckii, resulting in an imbalance in intracellular conditions associated with corresponding alterations in metabolic behaviors and fermentation efficiencies. UT-prestressed inoculum exhibited a 21.46% decrease in the membrane potential during the lag phase compared to untreated samples, with an intracellular pH of 5.68 ± 0.12, attributed to the lower activities of H+-ATPase and lactate dehydrogenase due to UT stress pretreatments. Comparative transcriptomic analysis revealed that UT prestress influenced the genes related to glycolysis, pyruvate metabolism, fatty acid synthesis, and ABC transport. The genes encoding 3-oxoacyl-[acyl-carrier-protein] reductases I, II, and III, CoA carboxylase, lactate dehydrogenase, pyruvate oxidase, glucose-6-phosphate isomerase, and glycerol-3-phosphate dehydrogenase were downregulated, thus identifying the relevance of the UT microstresses-downregulated absorption and utilization of carbohydrates with the attenuated fatty acid production and energy metabolisms. These findings could contribute to provide a better understanding of the inactivated effects on the post-acidification of L. delbrueckii by ultrasonic pretreatments, thus providing theoretical basis for the targeted optimization of acidification inhibition efficiencies for yogurt products during chilled preservation processes.


Assuntos
Fermentação , Perfilação da Expressão Gênica , Lactobacillus delbrueckii , Lactobacillus delbrueckii/metabolismo , Lactobacillus delbrueckii/genética , Concentração de Íons de Hidrogênio , Transcriptoma , Sonicação , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética
2.
Ultrason Sonochem ; 105: 106857, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38552299

RESUMO

This work investigated the effects of the combined use of thermosonication-preconditioned lactic acid bacteria (LAB) with the addition of ultrasound-assisted pineapple peel extracts (UU group) on the post-acidification potential, physicochemical and functional qualities of yogurt products, aimed at achieving prolonged preservation and enhancing functional attributes. Accordingly, the physical-chemical features, adhesion properties, and sensory profiles, acidification kinetics, the contents of major organic acids, and antioxidant activities of the differentially processed yogurts during refrigeration were characterized. Following a 14-day chilled storage process, UU group exhibited acidity levels of 0.5-2 oT lower than the control group and a higher lactose content of 0.07 mg/ml as well as unmodified adhesion potential, indicating that the proposed combination method efficiently inhibited post-acidification and delayed lactose metabolism without leading to significant impairment of the probiotic properties. The results of physicochemical analysis showed no significant changes in viscosity, hardness, and color of yogurt. Furthermore, the total phenolic content of UU-treated samples was 98 µg/mL, 1.78 times higher than that of the control, corresponding with the significantly lower IC50 values of DPPH and ABTS radical scavenging activities of the UU group than those of the control group. Observations by fluorescence inverted microscopy demonstrated the obvious adhesion phenomenon with no significant difference found among differentially prepared yogurts. The results of targeted metabolomics indicated the proposed combination strategy significantly modified the microbial metabolism, leading to the delayed utilization of lactose and the inhibited conversion into glucose during post-fermentation, as well as the decreased lactic acid production and a notable shift towards the formation of relatively weak acids such as succinic acid and citric acid. This study confirmed the feasibility of thermosonication-preconditioned LAB inocula, in combination with the use of natural active components from fruit processing byproducts, to alleviate post-acidification in yogurt and to enhance its antioxidant activities as well as simultaneously maintaining sensory features.


Assuntos
Ananas , Antioxidantes , Fermentação , Extratos Vegetais , Iogurte , Iogurte/microbiologia , Iogurte/análise , Ananas/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Antioxidantes/farmacologia , Sonicação , Temperatura , Concentração de Íons de Hidrogênio , Manipulação de Alimentos/métodos , Qualidade dos Alimentos
3.
Foods ; 13(2)2024 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-38254570

RESUMO

To better understand the textural transformation of Chinese dried noodles during the drying process, a convenient acoustic-force detection method was established. By comparing the breaking point, it was possible to determine the time-scale correlation between the force-displacement curves and acoustic spectrograms. The acoustic eigenvalues showed a consistent upward trend with the mechanical parameters during the drying process. With a wave crest reaching 152.8 dB and a signal maximum reaching 0.072, the structural stability of the dried noodles hardly induces a higher acoustic response. This suggests that the mechanical strength and rigidity of the dried noodles undergo minimal changes during this period. In comparison to the mechanical parameters, the acoustic eigenvalues accurately describe the changes in texture of dried noodles under various drying conditions, moreover, the sound threshold also provides a more effective response to the dried noodles' structural strength threshold. Therefore, the acoustic detection method can be applied to assist the conventional mechanical measurement in the field of the texture evaluation of dried food.

4.
Biomed Chromatogr ; 38(2): e5779, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38050189

RESUMO

To clarify the residue behavior and possible dietary risk of abamectin in fresh corn, bitter melon, and Fritillaria, a method was developed for the simultaneous determination of abamectin residues in fresh corn, bitter melon, and Fritillaria by QuEChERS (quick, easy, cheap, effective, rugged, safe) ultra-performance liquid chromatography-tandem mass spectrometry. The mean recovery of abamectin in fresh corn, bitter melon, and Fritillaria was 86.48%-107.80%, and the relative standard deviation was 2.07%-10.12%. The detection rates of abamectin residues in fresh corn, bitter melon, and Fritillaria were 62.50%, 87.50%, and 80.00%, respectively. The residues of abamectin in fresh corn, bitter melon, and Fritillaria were not more than 0.020, 0.019, and 0.087 mg/kg, respectively. Based on these results, dietary risk assessment showed that the risk content of abamectin residues in long- and short-term dietary exposure for Chinese consumers was 61.57% and 0.41%-1.11%, respectively, indicating that abamectin in fresh corn, bitter melon, and Fritillaria in the market would not pose a significant risk to consumers.


Assuntos
Fritillaria , Ivermectina/análogos & derivados , Momordica charantia , Resíduos de Praguicidas , Momordica charantia/química , Zea mays , Medição de Risco , Resíduos de Praguicidas/análise
5.
J Dairy Sci ; 107(5): 2760-2773, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38135047

RESUMO

This study aims to identify lactic acid bacteria (LAB) isolates possessing physiological characteristics suitable for use as probiotics in yogurt fermentation. Following acid and bile salt tolerance tests, Lactiplantibacillus plantarum (NUC08 and NUC101), Lacticaseibacillus rhamnosus (NUC55 and NUC201), and Lacticaseibacillus paracasei (NUC159, NUC216, and NUC351) were shortlisted based on intraspecies distribution for further evaluation. Their physiological probiotic properties, including transit tolerance, adhesion, autoaggregation, surface hydrophobicity, biofilm formation, and antibacterial activity, were assessed. Principal component analysis indicated that Lactiplantibacillus plantarum NUC08 was the preferred choice among the evaluated strains. Subsequent investigations revealed that co-culturing Lactiplantibacillus plantarum NUC08 with 2 yogurt starter strains resulted in a cooperative and synergistic effect, enhancing the growth of mixed strains and increasing their tolerance to simulated gastric and intestinal conditions. Additionally, when Vibrio harveyi bioluminescent reporter strain was used, the 3 cocultured strains cooperated to induce the activity of a quorum sensing (QS) molecule autoinducer-2 (AI-2), hinting a potential connection between phenotypic traits and QS in the cocultured strains. Importantly, LAB viable counts were significantly higher in yogurt co-fermented with Lactiplantibacillus plantarum NUC08, consistently throughout the storage period. In conclusion, the study demonstrates that the probiotic strain Lactiplantibacillus plantarum NUC08 can be employed in synergy with yogurt starter strains, affirming its potential for use in the development of functional fermented dairy products.


Assuntos
Produtos Fermentados do Leite , Lactobacillus plantarum , Probióticos , Animais , Iogurte/microbiologia , Lactobacillus plantarum/fisiologia , Lactobacillaceae
6.
Int J Mol Sci ; 24(7)2023 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-37047555

RESUMO

Flavonoids are easily destroyed and their activity lost during gastrointestinal digestion. Protein-based nanocomplexes, a delivery system that promotes nutrient stability and bioactivity, have received increasing attention in recent years. This study investigated the stability, inhibitory activity against α-glucosidase and interaction mechanisms of protein-based nanocomplexes combining whey protein isolate (WPI), soybean protein isolate (SPI) and bovine serum albumin (BSA) with flavonoids (F) from A. keiskei using spectrophotometry, fluorescence spectra and molecular docking approaches. The results show that the flavonoid content of WPI-F (23.17 ± 0.86 mg/g) was higher than those of SPI-F (19.41 ± 0.56 mg/g) and BSA-F (20.15 ± 0.62 mg/g) after simulated digestion in vitro. Furthermore, the inhibition rate of WPI-F (23.63 ± 0.02%) against α-glucosidase was also better than those of SPI-F (18.56 ± 0.02%) and BSA-F (21.62 ± 0.02%). The inhibition rate of WPI-F increased to nearly double that of F alone (12.43 ± 0.02%) (p < 0.05). Molecular docking results indicated that the protein-flavonoids (P-F) binding occurs primarily through hydrophobic forces, hydrogen bonds and ionic bonds. Thermodynamic analysis (ΔH > 0, ΔS > 0) indicated that the P-F interactions are predominantly hydrophobic forces. In addition, the absolute value of ΔG for WPI-F is greater (-30.22 ± 2.69 kJ mol-1), indicating that WPI-F releases more heat energy when synthesized and is more conducive to combination. This paper serves as a valuable reference for the stability and bioactivity of flavonoids from A. keiskei.


Assuntos
Angelica , Flavonoides , Flavonoides/farmacologia , Flavonoides/química , alfa-Glucosidases/metabolismo , Simulação de Acoplamento Molecular , Angelica/metabolismo , Ligação Proteica , Termodinâmica , Soroalbumina Bovina/química , Espectrometria de Fluorescência
7.
J Sci Food Agric ; 103(8): 3915-3925, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36335574

RESUMO

BACKGROUND: The traditional screening method for umami peptide, extracted from porcine bone, was labor-intensive and time-consuming. In this study, the rapid screening method and molecular mechanism of umami peptide was investigated. RESULTS: This article showed that a more precisely rapid screening method with composite machine learning and molecular docking was used to screen the potential umami peptide from porcine bone. As reference, 24 reported umami peptides were predicated by composite machine learning, with the accuracy of 86.7%. In this study, potential umami peptide sequences from porcine bone were screened by UMPred-FRL, Umami-MRNN Demo, and molecular docking was used to provide further screening. Finally, nine peptides were screened and verified as umami peptides by this method: LREY, HEAL, LAKVH, FQKVVA, HVKELE, AEVKKAP, EAVEKPQS, KALSEEL and KKMFETES. The hydrogen bonding was deemed to be the main interaction force with receptor T1R3, and domain binding sites were Ser146, His121 and Glu277. The result demonstrated the feasibility of machine learning assisted T1R1/T1R3 receptor for rapid screening umami peptides. The screening method would not only adapt to screen umami peptides from porcine bone but possibly applied for other sources. It also provided a reference for rapid screening of umami peptides. CONCLUSION: The manuscript lays a rapid screening method in screening umami peptide, and nine umami peptides from porcine bone were screened and identified. © 2022 Society of Chemical Industry.


Assuntos
Peptídeos , Receptores Acoplados a Proteínas G , Suínos , Simulação de Acoplamento Molecular , Receptores Acoplados a Proteínas G/metabolismo , Peptídeos/química , Sítios de Ligação , Ligação de Hidrogênio , Paladar , Animais
8.
Food Res Int ; 162(Pt B): 112127, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36461357

RESUMO

The potential of high-power ultrasonication (HPU) to enhance the physicochemical stability of bran-containing cereal products has been demonstrated, but the information concerning how the wholegrain volatilome and key quality-related chemical reactions evolve responding to HPU remains scarcely reported. The objective of this work was to examine the headspace volatile fingerprinting features of sonicated wholegrain brown rice (WBR; 400 W, 28 kHz, 30 min) following an accelerated storage testing (37 °C, 20 days), and simultaneously to identify the key chemical reactions induced by ultrasonication. A total of 70 aroma compounds were identified by the untargeted headspace GC-MS, including 9 alkanes, 6 alkenes, 15 aldehydes, 6 furans, 12 ketones, 9 alcohols and 13 miscellaneous compounds. Multivariate statistical analysis demonstrated that HPU pretreatments before a storage process significantly influenced the volatilome evolution, as revealed by a clear classification between sonicated and unsonicated grains. Supervised orthogonal partial least squares discriminant analysis identified the volatiles including acetic acid, pentanoic acid, hexanal, ethyl hexanoate and 2-pentyl-furan as HPU-related markers, inferring that HPU mainly modified the chemical reactions involving lipid decomposition, free fatty acids oxidation and esterification. This was further confirmed by targeted monitoring of lipid peroxidation products, with 13.22-14.84 % of MDA contents reduced (p < 0.05) in sonicated samples after storage. Besides, the ultrasonic effects resulted in a slight improvement of in vitro starch digestibility of WBR samples depending on the rice ecotype. This investigation demonstrated the potential of HPU pretreatments for prolonging the oxidative stability of WBR grains, without significantly compromising digestion properties.


Assuntos
Oryza , Amido , Grãos Integrais , Grão Comestível , Esterificação
9.
Nutrients ; 14(22)2022 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-36432413

RESUMO

Meat adulteration have become a global issue, which has increasingly raised concerns due to not only economic losses and religious issues, but also public safety and its negative effects on human health. Using optimal primers for seven target species, a multiplex PCR method was developed for the molecular authentication of camel, cattle, dog, pig, chicken, sheep and duck in one tube reaction. Species-specific amplification from the premixed total DNA of seven species was corroborated by DNA sequencing. The limit of detection (LOD) is as low as 0.025 ng DNA for the simultaneous identification of seven species in both raw and heat-processed meat or target meat: as little as 0.1% (w/w) of the total meat weight. This method is strongly reproducible even while exposed to intensively heat-processed meat and meat mixtures, which renders it able to trace meat origins in real-world foodstuffs based on the authenticity assessment of commercial meat samples. Therefore, this method is a powerful tool for the inspection of meat adulterants and has broad application prospects.


Assuntos
Temperatura Alta , Reação em Cadeia da Polimerase Multiplex , Bovinos , Suínos , Ovinos , Cães , Humanos , Animais , Reação em Cadeia da Polimerase Multiplex/métodos , Contaminação de Alimentos/análise , Carne/análise , DNA/análise
10.
Front Nutr ; 9: 979977, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36185690

RESUMO

Identification of meat authenticity is a matter of increasing concerns due to religious, economical, legal, and public health reasons. However, little is known about the inspection of eight meat species in one tube reaction due to technological challenge of multiplex polymerase chain reaction (PCR) techniques. Here, a developed multiplex PCR method can simultaneously authenticate eight meat species including ostrich (753 bp), cat (564 bp), goose (391 bp), duck (347 bp), chicken (268 bp), horse (227 bp), dog (190 bp), and sheep (131 bp). The detectable deoxyribonucleic acid (DNA) contents for each target species was as low as 0.01 ng in both raw and heat-treated meat or target meat down to 0.1% (w/w) of total meat weight reflecting high stability of the assay in heat processing condition, indicating that this method is adequate for tracing meat origin in real-world meat products, which has been further validated by authenticity assays of commercial meat products. Overall, this method is a powerful tool for accurate evaluation of meat origin with a good application foreground.

11.
Mikrochim Acta ; 189(11): 433, 2022 10 26.
Artigo em Inglês | MEDLINE | ID: mdl-36287256

RESUMO

A rapid, convenient, low-cost, and selective DNA isolation method was developed for identifying meat adulteration. A mesoporous metal organic framework (Meso-UIO-66)-coated solid phase microextraction system was employed as an isolation device to simplify DNA isolation into three steps (lysis, washing, and elution). Meso-UIO-66 was utilized as the adsorbent because of its positively charged surface, high chemical stability, and mesoporous structure. Meso-UIO-66 was characterized by scanning electron microscopy, transmission electron microscopy, powder X-ray diffraction, Fourier transform infrared spectroscopy, ultraviolet‒visible spectroscopy, X-ray photoelectron spectroscopy, and nitrogen adsorption-desorption tests. Parameters that affected DNA isolation were optimized. This method can be used to isolate and purify DNA from meat in 60 s, and the DNA concentration and purity are comparable to those of samples isolated with a commercial kit. Multiple DNA detection was achieved by coupling this method with the multiplex polymerase chain reaction technique, and the detection limit was lower than 1% (w/w).


Assuntos
Estruturas Metalorgânicas , Estruturas Metalorgânicas/química , Microextração em Fase Sólida/métodos , Pós , Limite de Detecção , Carne , Reação em Cadeia da Polimerase , DNA/genética , Nitrogênio
12.
Bioresour Technol ; 361: 127739, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35940323

RESUMO

This study explored the feasibility of thermosonication (TS)-prestressed inoculum with different fermentation patterns for regulating microbial (post)-fermentation acidification kinetics. Through a Box-Behnken design, stimulative (20 min, 400 W, 33 kHz, 25 °C) and inhibitive (10 min, 600 W, 33 kHz, 20 °C) effects on the acidification capability of Lactobacillus plantarum A3 were achieved without observing greatly activated/inactivated strains growth, further confirmed by lactose fermentation performed by Streptococcus thermophilus and Lactobacillus bulgaricus. Lactic acid was the major contributing factor responsible for TS-induced acidification modifications corresponding to the potential fluctuations of CoA biosynthesis, fatty acid degradation and chain elongation pathways to TS prestress. Microscopy observations and quantitative extracellular substance assays showed palpable stress disturbance on microbes, but causing insignificant effects on product characteristics. This investigation demonstrated the potential of controlled sonication prestress strategies to achieve dual engineering effects on microbial metabolic behavior, for alleviating post-acidification problem or enhancing process efficiencies.


Assuntos
Fermentação , Lactobacillales , Lactobacillus , Concentração de Íons de Hidrogênio , Cinética , Lactobacillus/metabolismo
13.
Front Nutr ; 9: 890537, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35811966

RESUMO

Frequent meat frauds have become a global issue because adulteration risks the food safety, breaches market rules, and even threatens public health. Multiplex PCR is considered to be a simple, fast, and inexpensive technique that can be applied for the identification of meat products in food industries. However, relatively less is known about a multiplex PCR method authenticating seven animal species simultaneously in one reaction due to technological challenge. Through screening new species-specific primers and optimizing PCR system, a heptaplex PCR method was established, which could simultaneously detect seven meat ingredients of camel (128 bp), pigeon (157 bp), chicken (220 bp), duck (272 bp), horse (314 bp), beef (434 bp), and pork (502 bp) in a single-tube reaction. DNA sequencing solidly validated that each set of primers specifically amplified target species from total DNA mixtures of seven meat species. The developed multiplex assay was stable and sensitive enough to detect 0.01-0.025 ng DNA from various meat treatments including raw, boiled, and autoclaved meat samples or target meat content of 0.1% total meat weight, suggesting the suitability of the heptaplex PCR technique for tracing target meats with high accuracy and precision. Most importantly, a market survey validated the availability of this multiplex PCR technique in real-world meat products with a good application foreground.

14.
Foods ; 11(11)2022 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-35681360

RESUMO

Lactobacillus are normal inhabitants of the gastrointestinal tract and confer a variety of health effects. Lipoteichoic acid (LTA), an amphiphilic substance located in the cell membrane, is a key molecule in probiotic-host crosstalk. Through the characterization of structural characteristics of LTA molecules derived from Lactobacillus plantarum A3, Lactobacillus reuteri DMSZ 8533, and Lactobacillus acidophilus CICC 6074, there exists some heterogeneity in LTA molecules, which perhaps contributes to the distinguishable adhesion properties of Lactobacillus strains based on fluorescence microscopy observations. In LPS-induced RAW 264.7 cells, LTAs derived from three Lactobacillus strains obviously alleviated inflammatory responses as evidenced by the altered inflammatory cytokine levels of TNF-α, IL-6, and IL-10. Western blotting demonstrated that L. reuteri LTA blocked LPS-triggered expression of the MAPK and NF-κB pathways. The findings further validated that LTA is an important effector molecule and deserves further consideration as an alternative therapeutic for ulcerative colitis treatment.

15.
Foods ; 11(12)2022 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-35741917

RESUMO

Lactobacillus acidophilus NCFM is widely used in the fermentation industry; using it as a freeze-dried powder can greatly reduce the costs associated with packaging and transportation, and even prolong the storage period. Previously published research has reported that the expression of galU (EC: 2.7.7.9) is significantly increased as a result of freezing and drying. Herein, we aimed to explore how galU plays an important role in improving the resistance of Lactobacillus acidophilus NCFM to freeze-drying. For this study, galU was first knocked out and then re-expressed in L. acidophilus NCFM to functionally characterize its role in the pertinent metabolic pathways. The knockout strain ΔgalU showed lactose/galactose deficiency and displayed irregular cell morphology, shortened cell length, thin and rough capsules, and abnormal cell division, and the progeny could not be separated. In the re-expression strain pgalU, these inhibited pathways were restored; moreover, the pgalU cells showed a strengthened cell wall and capsule, which enhanced their resistance to adverse environments. The pgalU cells showed GalU activity that was 229% higher than that shown by the wild-type strain, and the freeze-drying survival rate was 84%, this being 4.7 times higher than that of the wild-type strain. To summarize, expression of the galU gene can significantly enhance gene expression in galactose metabolic pathway and make the strain form a stronger cell wall and cell capsule and enhance the resistance of the bacteria to an adverse external environment, to improve the freeze-drying survival rate of L. acidophilus NCFM.

16.
Front Psychol ; 13: 881491, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35496234

RESUMO

This study attempted to explore the personality traits of higher achievers at the university level. The core objective of this investigation was to illustrate the nature of personality traits of the higher achievers' students. To study this phenomenon, a quantitative research approach was used. The students were chosen by using a purposive sampling technique and included 758 high achievers enrolled in various programs at the Chinese universities. Based on the Hexaco model of personality, a questionnaire was used to gather information from respondents as a research tool to examine the personality traits of position holders after an extensive review of the relevant literature. Tool validity was determined by following the face, content, construct (convergent and discriminant validity) validation process. This investigation concluded that honesty, emotionality, and openness to experience were very high among the higher achievers' students. Only honesty in female higher achievers' students was significantly high than male, remaining factors "extraversion, agreeableness, conscientiousness, and openness to experience" were significantly high among male higher achievers' students. Moreover, the higher achievers of science group students were more extraversion, agreeableness, and conscientiousness than arts group students. However, higher achievers in hostels were more emotional and agreeableness than the day scholars. Overall step-wise regression analysis, indicated that agreeableness and extraversion factor has significant influence on higher achievers.

17.
Front Nutr ; 9: 813962, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35399682

RESUMO

Frequent meat frauds have aroused significant social attention. The aim of this study is to construct a two-tube hexaplex polymerase chain reaction (PCR) method offering accurate molecular authentication of twelve meat species in actual adulteration event. Deoxyribonucleic acid (DNA) sequencing demonstrates that designed primers can specifically amplify target species from genomic DNA mixture of six species in each tube reaction, which showed 100% accuracy of horse (148 bp), pigeon (218 bp), camel (283 bp), rabbit (370 bp), ostrich (536 bp), and beef (610 bp) as well as turkey (124 bp), dog (149 bp), chicken (196 bp), duck (277 bp), cat (380 bp), and goose (468 bp). A species-specific primer pair produced the target band in the presence of target genomic DNA but not non-target species. Through multiplex PCR assays with serial concentration of the DNA mixture of six species in each PCR reaction, the detection limit (LOD) of the two-tube hexaplex PCR assay reached up to 0.05-0.1 ng. Using genomic DNA isolated from both boiled and microwave-cooked meat as templates, PCR amplification generated expected PCR products. These findings demonstrate that the proposed method is specific, sensitive and reproducible, and is adequate for food inspection. Most importantly, this method was successfully applied to detect meat frauds in commercial meat products. Therefore, this method is of great importance with a good application foreground.

18.
Appl Microbiol Biotechnol ; 106(7): 2481-2491, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35344091

RESUMO

Herein, two genes (LBA0625 and LBA1719) encoding UGPases (UDP-glucose pyrophosphorylase) in Lactobacillus acidophilus (L. acidophilus) were successfully transformed into Escherichia coli BL21 (DE3) to construct recombinant overexpressing strains (E-0625, E-1719) to investigate the biological characteristics of UGPase-0625 and UGPase-1719. The active sites, polysaccharide yield, and anti-freeze-drying stress of L. acidophilus ATCC4356 were also detected. UGPase-0625 and UGPase-1719 belong to the nucleotidyltransferase of stable hydrophilic proteins; contain 300 and 294 amino acids, respectively; and have 20 conserved active sites by prediction. Αlpha-helixes and random coils were the main secondary structures, which constituted the main skeleton of UGPases. The optimal mixture for the high catalytic activity of the two UGPases included 0.5 mM UDP-Glu (uridine diphosphate glucose) and Mg2+ at 37 °C, pH 10.0. By comparing the UGPase activities of the mutant strains with the original recombinant strains, A10, L130, and L263 were determined as the active sites of UGPase-0625 (P < 0.01) and A11, L130, and L263 were determined as the active sites of UGPase-1719 (P < 0.01). In addition, UGPase overexpression could increase the production of polysaccharides and the survival rates of recombinant bacteria after freeze-drying. This is the first study to determine the enzymatic properties, active sites, and structural simulation of UGPases from L. acidophilus, providing in-depth understanding of the biological characteristics of UGPases in lactic acid bacteria.Key points• We detected the biological characteristics of UGPases encoded by LBA0625 and LBA1719.• We identified UGPase-0625 and UGPase-1719 active sites.• UGPase overexpression elevates polysaccharide levels and post-freeze-drying survival.


Assuntos
Lactobacillus acidophilus , UTP-Glucose-1-Fosfato Uridililtransferase , Domínio Catalítico , Lactobacillus acidophilus/genética , Lactobacillus acidophilus/metabolismo , Estrutura Secundária de Proteína , UTP-Glucose-1-Fosfato Uridililtransferase/química , UTP-Glucose-1-Fosfato Uridililtransferase/genética , UTP-Glucose-1-Fosfato Uridililtransferase/metabolismo , Uridina Difosfato Glucose/metabolismo
19.
Compr Rev Food Sci Food Saf ; 21(2): 1462-1490, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35201672

RESUMO

Umami is an important element affecting food taste, and the development of umami peptides is a topic of interest in food-flavoring research. The existing technology used for traditional screening of umami peptides is time-consuming and labor-intensive, making it difficult to meet the requirements of high-throughput screening, which limits the rapid development of umami peptides. The difficulty in performing a standard measurement of umami intensity is another problem that restricts the development of umami peptides. The existing methods are not sensitive and specific, making it difficult to achieve a standard evaluation of umami taste. This review summarizes the umami receptors and umami peptides, focusing on the problems restricting the development of umami peptides, high-throughput screening, and establishment of evaluation standards. The rapid screening of umami peptides was realized based on molecular docking technology and a machine learning method, and the standard evaluation of umami could be realized with a bionic taste sensor. The progress of rapid screening and evaluation methods significantly promotes the study of umami peptides and increases its application in the seasoning industry.


Assuntos
Peptídeos , Paladar , Simulação de Acoplamento Molecular , Peptídeos/química
20.
Front Nutr ; 9: 1118900, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36712498

RESUMO

Lactobacillus strains fermentation of broccoli as a good source of bioactive peptides has not been fully elucidated. In this work, the peptide composition of broccoli fermented by L. plantarum A3 and L. rhamnosus ATCC7469 was analyzed by peptidomics to study the protein digestion patterns after fermentation by different strains. Results showed that water-soluble proteins such as rubisco were abundant sources of peptides, which triggered the sustained release of peptides as the main target of hydrolysis. In addition, 17 novel anti-inflammatory peptides were identified by virtual screening. Among them, SIWYGPDRP had the strongest ability to inhibit the release of NO from inflammatory cells at a concentration of 25 µM with an inhibition rate of 52.32 ± 1.48%. RFR and KASFAFAGL had the strongest inhibitory effects on the secretion of TNF-α and IL-6, respectively. At a concentration of 25 µM, the corresponding inhibition rates were 74.61 ± 1.68% and 29.84 ± 0.63%, respectively. Molecular docking results showed that 17 peptides formed hydrogen bonds and hydrophobic interactions with inducible nitric oxide synthase (iNOS). This study is conducive to the high-value utilization of broccoli and reduction of the antibiotic use.

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