Antioxidant packaging films developed by in-situ cross-linking chitosan with dialdehyde starch-catechin conjugates.

In this study, four dialdehyde starch-catechin (DAS-catechin) conjugates were prepared by conjugating (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG) and (-)-epigallocatechin gallate (EGCG) with dialdehyde starch. Then, DAS-catechin conjugates were used as antioxidant and cross-linking agents to produce chitosan (CS) based antioxidant packaging films. The functionality of CS/DAS-catechin conjugate films was determined. Results showed four DAS-catechin conjugates formed Schiff-base linkages and hydrogen bond interactions with CS, resulting in improved film uniformity. Compared with plain CS film, CS/DAS-catechin conjugate films owned higher UV-vis light, water vapor and oxygen barrier ability, lower swelling degree, and stronger tensile strength, thermal stability and antioxidant activity. The cross-linking between CS and DAS-catechin conjugates delayed the biodegradable process of CS film. CS/DAS-catechin conjugate films showed good performance on inhibiting sunflower seed oil oxidation. Notably, CS/DAS-ECG conjugate film had the highest oxygen barrier, mechanical and antioxidant performances among four CS/DAS-catechin conjugate films. Therefore, CS/DAS-ECG conjugate film is an antioxidant packaging candidate for edible oil.

Structure, stability and antioxidant activity of dialdehyde starch grafted with epicatechin, epicatechin gallate, epigallocatechin and epigallocatechin gallate.

BACKGROUND: Catechins, a member of the flavonoids, exist widely in teas, and have health benefits. However, catechins have poor stability, which greatly limits their application. In order to improve the stability of catechins, different catechins including (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin (EGC) and (-)-epigallocatechin gallate (EGCG) were conjugated onto dialdehyde starch by acid-mediated coupling method. The structure, stability and antioxidant activity of dialdehyde starch-catechin conjugates were determined. RESULTS: Thin-layer chromatography and ultraviolet-visible spectroscopy, fluorescence, nuclear magnetic resonance and infrared spectra revealed that catechins were successfully conjugated onto dialdehyde starch, coupling between 6-H/8-H of catechins' A ring and dialdehyde starch's aldehyde groups. The conjugates presented an amorphous structure and sheet-like and/or blocky morphologies. As compared to dialdehyde starch, the conjugates showed enhanced thermal stability. Furthermore, the stability of catechins in pH 7.4 phosphate-buffered saline was improved after conjugating onto dialdehyde starch. The conjugates exhibited significantly higher antioxidant activities than dialdehyde starch, decreasing in the following order: dialdehyde starch-ECG, dialdehyde starch-EGCG, dialdehyde starch-EC, dialdehyde starch-EGC and dialdehyde starch. CONCLUSION: Dialdehyde starch-catechin conjugates have great potential as stable antioxidant agents. © 2022 Society of Chemical Industry.

Effect of chitosan/starch aldehyde-catechin conjugate composite coating on the quality and shelf life of fresh pork loins.

BACKGROUND: Fresh pork is susceptible to oxidation and spoilage. Edible coating containing antioxidant and antimicrobial agents can create moisture and oxygen barriers around pork and inhibit oxidation and microbial growth in the pork. In this study, chitosan in combination with starch aldehyde-catechin conjugate (SACC) was used as a novel edible coating material for preserving fresh pork loins at chilled storage (4 ± 1 °C) for 14 days. Effect of chitosan/SACC composite coating on the quality of pork loins including weight loss, colour, pH value, microbial spoilage, lipid oxidation, protein oxidation, texture and sensory attributes during chilled storage was determined. RESULTS: Chitosan and SACC had synergistic antioxidant and antimicrobial actions. As compared with uncoated and chitosan coated pork loins, chitosan/SACC coated pork loins showed lower weight loss (7.16%), pH value (5.99), total viable count (7.11 log CFU g-1 ), total volatile base nitrogen content (130.2 mg kg-1 ), lipid oxidation level (0.47 mg malondialdehyde kg-1 ), protein oxidation level (0.047 mmol free thiol group g-1 ) and shear force (27.40 N) on day 14. Meanwhile, chitosan/SACC composite coating effectively maintained the colour, micro-structure and sensory attributes of pork loins throughout chilled storage period. The shelf life of pork loins was extended from 8 days (uncoated samples) to 14 days by chitosan/SACC composite coating. CONCLUSION: Chitosan/SACC composite coating effectively retarded the oxidation and spoilage of pork loins during chilled storage. © 2022 Society of Chemical Industry.

Horseradish peroxidase catalyzed grafting of chitosan oligosaccharide with different flavonols: structures, antioxidant activity and edible coating application.

BACKGROUND: Enzymatic catalyzed grafting of oligosaccharides with polyphenols is a safe and environmentally friendly approach to simultaneously enhance the bioactivity of oligosaccharides and the solubility of polyphenols. In this study, chitosan oligosaccharide (COS) was grafted with three different flavonols including myricetin (MYR), quercetin (QUE) and kaempferol (KAE) by horseradish peroxidase (HRP) catalysis. The structures, antioxidant activity and edible coating application of COS-flavonol conjugates were investigated. RESULTS: The total phenol content of COS-MYR, COS-QUE and COS-KAE conjugates was 59.89, 68.37 and 53.77 mg gallic acid equivalents g-1 , respectively. Thin layer chromatography showed the conjugates did not contain ungrafted flavonols. COS-flavonol conjugates showed ultraviolet absorption peak at about 294 nm, corresponding to the A-ring of flavonols. Fourier-transform infrared spectra of conjugates confirmed the formation of Schiff-base and Michael-addition products. The proton-nuclear magnetic resonance spectrum of COS-KAE conjugate exhibited phenyl proton signals of KAE. X-ray diffraction patterns of conjugates showed some diffraction peaks of flavonols. COS-flavonol conjugates presented rough and porous morphologies with sheet-like and/or blocky structures. The conjugates showed higher water solubility, free radical scavenging activity and reducing power than flavonols. Moreover, fish gelatin/COS-flavonol conjugate coatings effectively prolonged the shelf life of refrigerated largemouth bass (Micropterus salmoides) fillets from 5 days to 7-8 days. CONCLUSION: COS-flavonol conjugates prepared by HRP catalysis have great potentials as novel antioxidant agents. © 2022 Society of Chemical Industry.

Highly efficient synthesis and characterization of starch aldehyde-catechin conjugate with potent antioxidant activity.

In this study, cassava starch aldehyde was functionalized with catechin through acid catalyzed condensation reaction. The structural characterization, stability and antioxidant activity of starch aldehyde-catechin conjugates were investigated. Thin layer chromatography revealed the conjugates did not contain free catechin. UV-vis spectra of the conjugates exhibited an absorption band at 280 nm, attributing to the B-ring of catechin moiety. Fourier-transform infrared and proton nuclear magnetic resonance spectroscopy demonstrated the conjugation occurred between the H-6/H-8 of catechin A-ring and the aldehyde groups of starch aldehyde. X-ray diffraction pattern indicated that the conjugates had an amorphous structure. Scanning electron microscopy showed the conjugates were fragmentary slices with rough surfaces. Notably, the conjugates were more stable than catechin in phosphate buffered saline (pH 7.4). In addition, the conjugates could not be digested in simulated saliva, gastric and small intestinal juices. The reducing power and free radical scavenging activity of starch aldehyde were remarkably elevated by conjugating with catechin. Meanwhile, the conjugates were non-cytotoxic to RAW264.7 mouse macrophage cells and possessed higher resistant starch contents than starch. Our results suggest starch aldehyde-catechin conjugates can be used as antioxidants in food industry.

Comparison of the physical and functional properties of starch/polyvinyl alcohol films containing anthocyanins and/or betacyanins.

Anthocyanins and betacyanins are two different types of natural pigments. In this study, active and intelligent packaging were prepared by adding anthocyanins or betacyanins or anthocyanins/betacyanins mixture (in the weight ratios of 3:1, 1:1 and 1:3) into starch/polyvinyl alcohol (PVA) films. The physical and functional properties of developed films were compared. Results showed anthocyanins were more pH-sensitive than betacyanins. However, betacyanins were more beneficial to improve the compactness of the films and had stronger hydrogen bonding interactions with film matrix as compared with anthocyanins. The addition of anthocyanins and/or betacyanins slightly decreased the crystallinity of the films but significantly increased the ultraviolet-visible light barrier, water vapor barrier, antioxidant, antimicrobial, and ammonia-sensitive properties of the films. Moreover, the barrier, antioxidant and antimicrobial properties of the films increased with the increase of betacyanins proportion. Amongst different films, the film containing anthocyanins/betacyanins mixture in the weight ratio of 1:3 presented obvious color changes when the film was utilized to monitor the freshness of pork. Our results suggested betacyanins was more beneficial to enhance the barrier, antioxidant and antimicrobial properties of starch/PVA films. Starch/PVA films simultaneously added with anthocyanins and betacyanins were more suitable to monitor the freshness of pork.

Development of multifunctional food packaging by incorporating betalains from vegetable amaranth (Amaranthus tricolor L.) into quaternary ammonium chitosan/fish gelatin blend films.

Multifunctional food packaging was prepared by incorporating betalains-rich vegetable amaranth (Amaranthus tricolor L.) extract into quaternary ammonium chitosan (QC)/fish gelatin (FG) blend films. The structures, physical and functional properties of the films with and without amaranth extract (AE) were determined. Results showed betalains-rich AE was pH-sensitive and could change its colors under alkaline conditions. Structural characterization revealed the interactions between AE and film matrix were based on hydrogen bonds, which influenced the compactness of the films. Notably, AE significantly enhanced the thickness (from 0.076 to 0.089 mm), UV-vis light barrier ability, elongation at break (from 66.7% to 88.8%), 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (from 6.8% to 95.8% at 5 mg/mL), antimicrobial activity against four foodborne pathogens (Escherichia coli, Staphylococcus aureus, Salmonella typhimurium and Listeria monocytogenes) and ammonia sensitivity of the films. However, AE remarkably reduced the water vapor permeability (from 34.9 × 10-11 to 26.4 × 10-11 g m-1 s-1 Pa-1) of the films. Notably, the films containing 5 wt% and 10 wt% of AE were suitable to monitor the freshness of shrimp. Our results suggested QC/FG films with betalains-rich AE could be used as antioxidant, antimicrobial and intelligent packaging in food industry.

Recent advances in phenolic-protein conjugates: synthesis, characterization, biological activities and potential applications.

Proteins and phenolic compounds are two types of food ingredients with distinct functionalities. In the past decade, many attempts have been made to conjugate phenolic compounds with proteins through covalent linkages. Four types of conjugation reactions including alkaline, free radical mediated grafting, enzyme catalyzed grafting and chemical coupling methods are frequently used to synthesize phenolic-protein conjugates. The synthesized phenolic-protein conjugates can be well characterized by several different instrumental methods, such as UV spectroscopy, Fourier transform infrared spectroscopy, fluorescence spectroscopy, circular dichroism, mass spectroscopy, sodium dodecyl sulfate polyacrylamide gel electrophoresis and differential scanning calorimetry. Importantly, phenolic-protein conjugates exhibit improved biological properties (e.g. antioxidant, anticancer and antimicrobial activities) as compared with native proteins. Moreover, the applications of native proteins can be greatly widened by conjugation with phenolic compounds. Phenolic-protein conjugates have been developed as antioxidant emulsions for nutraceutical delivery, edible films for food packaging, stabilizers for metal nanoparticles, and hydrogels and nanoparticles for controlled drug release. In this review, recent advances in the synthesis, characterization, biological properties and potential applications of phenolic-protein conjugates were summarized.

[Expression and characterization of the FABP and Eg95 fusion gene from Echinococcus granulosus].

OBJECTIVE: To construct and express a fusion gene of fatty acid binding protein (FABP) with Eg95 which are protective antigen genes of Echinococcus granulosus, and investigate the immunological characteristics of the recombinant protein. METHOD: Using cDNA fragments encoding FABP and Eg95 genes from E. granulosus Qinghai sheep strain as templates, a fusion gene FABP.Eg95 was amplified by asymmetric polymerase chain reaction th-rough a linking sequence encoding four glycine residues. The PCR products of fusion gene were subcloned into the pET28a (+) vector. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3) cells and induced with IPTG. The recombinant protein was detected by SDS-PAGE and purified by Ni-IDA affinity chromatography, and its immunogenicity was analyzed by Western blotting. Results The fusion gene length was about 795 bp. Double enzyme digestion analysis and DNA sequencing confirmed that the fusion gene was cloned into pET-28a (+). The recombinant protein (Mr 31,000) was expressed in inclusion body in E. coli expression system, and was purified by Ni-IDA affinity chromatography. Western blotting analysis testified that the recombinant protein could be recognized by sera of cystic echinococcosis patients, but not by sera of healthy persons and schistosomiasis patients. CONCLUSION: The FABP.Eg95 fusion gene has been constructed, and the purified recombinant protein has been confirmed with immunogenicity.

Molecular cloning and identification of a novel Clonorchis sinensis gene encoding a tegumental protein.

The tegumental membrane of platyhelminth parasites is of crucial importance for modulation of the host response and parasite survival. A cDNA encoding a novel tegumental protein 20.8 kDa (TP20.8) was found by large-scale sequencing of a Clonorchis sinensis cDNA library. This new cDNA was 755 bp long containing an open reading frame of 555 bp, which encoded a 20.8-kDa protein with an isoelectric point of 4.33. The deduced amino acid sequence exhibits 40 and 37% identity to Schistosoma japonicum sj20.8 and Schistosoma mansoni Sm 20.8, respectively. TP20.8 transcripts were detected in the adult worm and metacercariae cDNA libraries of C. sinensis but not in the egg. Recombinant C. sinensis TP20.8 protein was produced and purified from Escherichia coli BL21. Using specific anti-recombinant TP20.8 protein sera, the TP20.8 protein was immunohistochemically localized to the outer-surface membrane of C. sinensis. The specificity and sensitivity of the recombinant antigen for serologic diagnosis was assessed by enzyme-linked immunosorbent assay using serum from 100 patients with clonorchiasis, 20 patients with schistosomiasis, and 30 negative controls. The sensitivity was 68%, and the specificity was 84%. The antigen was less useful for the serodiagnosis of clonorchiasis with IgG.