A cysteine peptidase was previously identified from germinated Moringa oleifera seeds, but its milk-clotting properties on bovine caseins was still unclear. In this study, this novel cysteine ...peptidase (MoCP) showed preferential activity on κ-casein (κ-CN), with greater hydrolytic activity compared with calf rennet, whereas weak hydrolysis of α-casein and β-casein made MoCP suitable for application in cheesemaking and may yield various functional peptides. All 3 evaluated caseins were hydrolyzed to form relatively stable peptide bands within 3 h of proteolysis with MoCP. Cleavage sites were determined by gel electrophoresis, liquid chromatography mass spectrometry/mass spectrometry, and peptide sequencing, which revealed that cleavage of κ-CN by MoCP occurred at residue Ile129-Pro130 and generated a 14,895.37-Da peptide. The flocculation reaction between MoCP and κ-CN determined by 3-dimensional microscopy with super-depth of field revealed that the initial 30 min of reaction were key for milk coagulation, which may affect curd yield. Overall, the findings presented herein suggest that the cysteine peptidase from germinated M. oleifera seeds can be considered a promising plant-derived rennet alternative for use in cheese manufacture.
As primary coffee by-products, Arabica coffee husks are largely discarded during coffee-drying, posing a serious environmental threat. However, coffee husks could be used as potential material for ...extracting pectin polysaccharides, with high bioactivities and excellent processing properties. Thus, the present study aimed to extract the pectin polysaccharide from Arabica coffee husk(s) (CHP). The CHP yield was calculated after vacuum freeze-drying, and its average molecular weight (Mw) was detected by gel permeation chromatography (GPC). The structural characteristics of CHP were determined by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), proton nuclear magnetic resonance (
H NMR), and scanning electron microscopy (SEM). Additionally, the rheological and antioxidant properties of CHP and the inhibition capacities of advanced glycation end products (AGEs) with different concentrations were evaluated. The interaction mechanisms between galacturonic acid (GalA) and the AGE receptor were analyzed using molecular docking. The results demonstrated that the CHP yield was 19.13 ± 0.85%, and its Mw was 1.04 × 10
Da. The results of the structural characteristics results revealed that CHP was an amorphous and low-methoxyl pectic polysaccharide linked with an α-(1→6) glycosidic bond, and mainly composed of rhamnose (Rha, 2.55%), galacturonic acid (GalA, 45.01%), β-N-acetyl glucosamine (GlcNAc, 5.17%), glucose (Glc, 32.29%), galactose (Gal, 6.80%), xylose (Xyl, 0.76%), and arabinose (Ara, 7.42%). The surface microstructure of CHP was rough with cracks, and its aqueous belonged to non-Newtonian fluid with a higher elastic modulus (G'). Furthermore, the results of the antioxidant properties indicated that CHP possessed vigorous antioxidant activities in a dose manner, and the inhibition capacities of AGEs reached their highest of 66.0 ± 0.35% at 1.5 mg/mL of CHP. The molecular docking prediction demonstrated that GalA had a good affinity toward AGE receptors by -6.20 kcal/mol of binding energy. Overall, the study results provide a theoretical basis for broadening the application of CHP in the food industry.
The peptide MOp2 obtained from
Moringa oleifera
seeds showed good antimicrobial activity. However, the stability of its activity has not yet been studied. In the present study, MOp2-loaded thiolated ...chitosan-stabilized (CMOp2) Pickering emulsion was prepared and applied to prolong the shelf life of grass carp. The encapsulation rate of MOp2 was 57.7% in CMOp2. In addition, the effects of different concentrations of CMOp2 solid particles and pH on droplet size, zeta optional and storage stability of Pickering emulsions were evaluated; the best condition for preparing Pickering emulsion through experiment was 1.75% CMOp2 solid particles at pH 9.5. Moreover, morphological observations and rheological analysis indicated that Pickering emulsions were considered a water-in-oil emulsion with typical non-Newtonian fluid characteristics. Furthermore, the prepared Pickering emulsion could significantly inhibit the growth of
Escherichia coli
and
Staphylococcus aureus
. Besides, Pickering emulsion effectively prevented spoilage of grass carp, and the Pickering emulsion-treated group reduced its pH, TVB-N and color values, inhibited microbial growth, and extended shelf life to 9 day at the storage of 4 °C. Overall, the present findings provide a reference for the application of MOp2-loaded Pickering emulsions in food preservation.
Image 1
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MOp2-loaded Pickering emulsion stabilized with thiolated chitosan was prepared.
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Pickering emulsion as a water-in-oil emulsion had typical non-Newtonian behavior.
•
Pickering emulsion showed antimicrobial activity against
E. coli
and
S. aureus.
•
Prepared Pickering emulsion was applied to the preservation of grass carp.
This study aimed to compare chemical compositions, antioxidant activities, and acetylcholinesterase inhibitory activities of coffee flowers (ACF) and coffee leaves (ACL) with green coffee beans ...(ACGB) of Coffea Arabica L. The chemical compositions were determined by employing high‐performance liquid chromatography–mass spectroscopy (HPLC–MS) and gas chromatography–mass spectroscopy (GC–MS) techniques. Antioxidant effects of the components were evaluated using DPPH and ABTS radical scavenging assays, and the ferric reducing antioxidant power (FRAP) assay. Their acetylcholinesterase inhibitory activities were also evaluated. The coffee sample extracts contained a total of 214 components identified by HPLC‐MS and belonged to 12 classes (such as nucleotides and amino acids and their derivatives, tannins, flavonoids, alkaloids, benzene, phenylpropanoids, and lipids.), where phenylpropanoids were the dominant component (>30%). The contents of flavonoids, alkaloids, saccharides, and carboxylic acid and its derivatives in ACF and ACL varied significantly (p < .05) compared to similar components in ACGB. Meanwhile, 30 differentially changed chemical compositions (variable importance in projection VIP > 1, p < .01 and fold change FC > 4, or <0.25), that determine the difference in characteristics, were confirmed in the three coffee samples. Furthermore, among 25 volatile chemical components identified by GC–MS, caffeine, n‐hexadecanoic acid, 2,2′‐methylenebis6‐(1,1‐dimethylethyl)‐4‐methyl‐phenol, and quinic acid were common in these samples with caffeine being the highest in percentage. In addition, ACL showed the significantly highest (p < .05) DPPH radical scavenging capacity with IC50 value of 0.491 ± 0.148 mg/ml, and acetylcholinesterase inhibitory activity with inhibition ratio 25.18 ± 2.96%, whereas ACF showed the significantly highest (p < .05) ABTS radical scavenging activity with 36.413 ± 1.523 mmol trolox/g Ex. The results suggested that ACL and ACF had potential values as novel foods in the future.
The chemical compositions of coffee flowers and leaves Arabic Coffea from Yunnan province, China were compared with green coffee beans by HPLC–MS and GC–MS. Antioxidant and acetylcholinesterase inhibitory activities were higher than green coffee beans in vitro. Thirty differentially changed chemical compositions were confirmed.
The primary processing method of coffee plays a crucial role in determining its flavor profile. In this study, roasted coffee beans were subjected to three primary processing methods, i.e., natural ...processing (SC), washed processing (WC), and honey processing (MC), that were analyzed by LC-MS/MS and GC-MS metabolomics. Additionally, sensory evaluation was conducted by the Specialty Coffee Association of America (SCAA) to assess coffee flavor characteristics. The results showed that 2642 non-volatile compounds and 176 volatile compounds were detected across the three primary processing methods. Furthermore, significant differentially changed non-volatile compounds (DCnVCs) and volatile compounds (DCVCs) were detected among SC/WC (137 non-volatile compounds; 32 volatile compounds), MC/SC (103 non-volatile compounds; 25 volatile compounds), and MC/WC (20 non-volatile compounds; 9 volatile compounds). Notable compounds, such as lichenin, 6-gingerdiol 5-acetate, 3-fluoro-2-hydroxyquinoline, and 4-(4-butyl-2,5-dioxo-3-methyl-3-phenyl-1-pyrrolidiny)benzenesulfonamide, were identified as important DCnVCs, while ethyl alpha-D-glucopyranoside, 2,3-butanediol, maltol, and pentane-1,2,5-triol were identified as significant DCVCs in SC/WC. In MC/SC, 3-fluoro-2-hydroxyquinoline, etimicin, lichenin, and imazamox were important DCnVCs, whereas ethyl alpha-D-glucopyranoside, 2-pyrrolidinone, furfuryl alcohol, and pentane-1,2,5-triol were import DCVCs. Lastly, MC/WC samples exhibited notable DCnVCS, such as (S)-2-hydroxy-2-phenylacetonitrile O-b-D-apiosyl-1->2-b-D-glucoside, CMP-2-aminoethyphosphonate, talipexole, and neoconvallatoxoloside, along with DCVCS including citric acid, mannonic acid, gamma-lactone, 3-(1-hydroxy-1-methylethyl)benzonitrile, and maltol. Therefore, the primary processing method was a useful influence factor for coffee compositions.
The metabolic modulation of major flavor precursors during coffee cherry ripening is critical for the characteristic coffee flavor formation. However, the formation mechanism of flavor precursors ...during coffee cherry ripening remains unknown. In the present study, a colorimeter was employed to distinguish different maturity stages of coffee cherry based on the coffee cherry skin colors, and proteomics and metabolomics profiles were integrated to comprehensively investigate the flavor precursor dynamics involved in Arabica coffee cherry ripening. The data obtained in the present study provide an integral view of the critical pathways involved in flavor precursor changes during coffee cherry ripening. Moreover, the contributions of critical events in regulating the development of flavor precursors during the four ripening stages of coffee cherries, including the biosynthesis and metabolism pathways of organic acids, amino acids, flavonoids, and sugars, are discussed. Overall, a total of 456 difference express metabolites were selected, and they were identified as being concentrated in the four maturity stages of coffee cherries; furthermore, 76 crucial enzymes from the biosynthesis and metabolism of sugars, organic acids, amino acids, and flavonoids contributed to flavor precursor formation. Among these enzymes, 45 difference express proteins that could regulate 40 primary amino acids and organic acids flavor precursors were confirmed. This confirmation indicates that the metabolic pathways of amino acids and organic acids played a significant role in the flavor formation of Arabica coffee cherries during ripening. These results provide new insights into the protease modulation of flavor precursor changes in Arabica coffee cherry ripening.
Two series of novel fraxinellone-based thioethers containing 1,3,4-oxadiazole moiety were prepared as insecticidal agents against the oriental armyworm, Mythimna separata Walker. The structural ...assignment was based on the spectroscopic and X-ray analysis data. Among all the target compounds, compounds 4b, 4k, 5b, 5j and 5k exhibited more potent insecticidal activity with final mortality rates (FMRs) of more than 65%, especially 4k with the FMR of 75.9%, when compared with toosendanin. Some interesting results of structure–activity relationships are also discussed.
The malignant mesothelioma (MM) survival rate has been hampered by the lack of efficient and accurate early detection methods. The immune system may detect the early changes of tumor progression by ...responding with tumor-associated autoantibody production. Hence, in this study, we translated the humoral immune response to cancer proteins into a potential blood test for MM.
A T7 phage MM cDNA library was constructed using MM tumor tissues and biopanned for tumor-associated antigens (TAAs) using pooled MM patient and normal serum samples. About 1008 individual phage TAA clones from the biopanned library were subjected to protein microarray construction and tested with 53 MM and 52 control serum samples as a training group. Nine candidate autoantibody markers were selected from the training group using Tclass system and logistic regression statistical analysis, which achieved 94.3% sensitivity and 90.4% specificity with an AUC value of 0.89 in receiver operating characteristic analysis. The classifier was further evaluated with 50 patient and 50 normal serum samples as an independent blind validation, and the sensitivity of 86.0% and the specificity of 86.0% were obtained with an AUC of 0.82. Sequencing and BLASTN analysis of the classifier revealed that five of these nine candidate markers were found to have strong homology to cancer related proteins (PDIA6, MEG3, SDCCAG3, IGHG3, IGHG1).
Our results indicated that using a panel of 9 autoantibody markers presented a promising accuracy for MM detection. Although the results need further validation in high-risk groups, they provided the potentials in developing a serum-based assay for MM diagnosis.
To reduce pollution caused by traditional plastic packaging and preparation of silver nanoparticles (AgNPs), this work aims to develop biological macromolecular packaging films with green synthesized ...AgNPs. In this study, a novel P. cocos polysaccharide (PCP) with a unique monosaccharide composition was extracted from Poria cocos (Schw.) Wolf. Then, this polysaccharide containing 24.68 % rhamnose was used as a stabilizer for the green synthesis of PCP-AgNPs for the first time. PCP-AgNPs exhibited excellent antibacterial activity against P. aeruginosa, E. coli, and S. aureus, with the highest antibacterial activity against E. coli (inhibition zone diameter = 11.14 ± 0.79 mm). Subsequently, PCP-AgNPs/chitosan (CS) film was successfully prepared by incorporating PCP-AgNPs into the CS film solution. Several experiments demonstrated that the addition of this nanomaterial promoted the formation of noncovalent interactions between CS and PCP-AgNPs, resulting in a more regular and denser film. Compared to the CS film and control group, the PCP-AgNPs/CS film significantly maintained the quality indexes of strawberries. Therefore, this composite film successfully extended the shelf life of strawberries. Regarding safety, these packaging films were not cytotoxic toward RAW264.7 cells. In conclusion, the environmentally friendly PCP-AgNPs/CS film has the potential to replace some traditional food packaging materials.
•The PCP-AgNPs were green synthesized by using the P. cocos polysaccharide (PCP) as green stabilizer for the first time.•PCP-AgNPs were added to CS for developing packaging film (PCP-AgNPs/CS) with good packaging properties.•PCP-AgNPs/CS film delayed the degradation of multiple sensory and physicochemical indexes of strawberry.•PCP-AgNPs/CS film show promising applications in food preservation.
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•A novel antimicrobial peptide (MOp3) was isolated and identified from M. oleifera seeds.•MOp3 exhibited potent antimicrobial activity and negligible hemolytic activity.•MOp3 induced ...irreversible damage on cell membrane of Staphylococcus aureus.•MOp3 interacted with DNA gyrase and dihydrofolate reductase of S. aureus.
This study aimed to characterize a novel antimicrobial peptide (AMP) obtained from Moringa oleifera seed protein hydrolysates. Cell membrane chromatography and live bacteria adsorption were combined into a single step to efficiently isolate the active fraction of the AMP. Five peptides were identified by LC-MS/MS, among which the MCNDCGA peptide (termed MOp3) showed the greatest inhibitory effect against Staphylococcus aureus minimum inhibitory concentration (MIC): 2 mg/mL. MOp3 was identified as a hydrophobic anionic AMP rich in β-sheet structures with negligible hemolytic activity at 2.0 × MIC. MOp3 had good tolerance to salt solutions at 5 % and pH range 6.0–8.0, but was sensitive to high temperatures (>100 °C) and acid protease. Microscopic observation further revealed that MOp3 induced irreversible damage onto the cell membrane of S. aureus and interacted with dihydrofolate reductase and DNA gyrase by hydrogen bonding and hydrophobic interaction. These findings highlight the potential application of a new antimicrobial agent against S. aureus in the food industry.
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