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Evaluation of yellow mealworm (Tenebrio molitor) larvae protein extract as a cryoprotectant for frozen Pacific white shrimp
(2026-06-15)
Kittiphattanabawon, Phanat
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Sriket, Chodsana
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Nalinanon, Sitthipong
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Phaonakrop, Narumon
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Roytrakul, Sittiruk
This study evaluated the application of yellow mealworm ( Tenebrio molitor ) larvae protein extract (YMPE) as a clean-label cryoprotectant for frozen Pacific white shrimp ( Litopenaeus vannamei ), with emphasis on its underlying mechanism. Proteomic profiling identified 12 antifreeze protein-like peptides containing conserved ice-binding motifs (TCTxSxxCxxAx), supporting the cryoprotective potential of YMPE. During dose optimization, shrimp treated with 0.5% YMPE exhibited the highest weight gain (5.57%) and lowest freezable water content (59.67%) (P < 0.05). The cryoprotective efficacy of YMPE was further evaluated over five freeze–thaw cycles in comparison with distilled water and mixed phosphate. YMPE significantly mitigated quality deterioration by enhancing water retention, preserving protein functionality (higher Ca2+-ATPase activity), and reducing lysosomal enzyme leakage relative to the control (P < 0.05). Microstructural analysis revealed reduced muscle fiber disruption, characterized by fewer inter-fiber gaps. Mechanistically, YMPE may exert cryoprotective effects through a multimodal mechanism involving ice recrystallization inhibition and formation of a protective protein matrix. Although less effective than mixed phosphate, YMPE consistently improved all quality parameters compared with the control. These findings highlight the potential of insect-derived protein extracts as a novel, biologically based alternative to phosphate additives in seafood processing and provide a foundation for future industrial application.
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Genomic Assessment, Metabolic Profile Mapping, and Anti-Helicobacter pylori Activity of Lactococcus lactis SK2-659 from Thai Fermented Green Mustard (Pak-kad-dong)
(2026-01-01)
Kingkaew, Engkarat
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Woraprayote, Weerapong
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Sitdhipol, Jaruwan
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Vilaichone, Ratha Korn
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Visessanguan, Wonnop
Probiotics play crucial roles in promoting gut health, enhancing immunity, and combating pathogenic microorganisms, with increasing interest in their applications in the food and therapeutic industries. Lactococcus lactis, a well-known lactic acid bacterium, has emerged as a promising candidate owing to its probiotic traits and safety profile. In this study, we investigated the probiotic potential and genomic profile of Lactococcus lactis SK2-659, a strain isolated from Thai fermented green mustard (Pak-kad-dong). Genomic analysis revealed numerous genes associated with probiotic traits, including stress tolerance, adhesion, and antimicrobial activity, with a particular focus on bacteriocin SK2-659, which was effective against pathogenic bacteria such as Helicobacter pylori. The bacteriocin produced by L. lactis SK2-659, identified as nisin Z, disrupts bacterial membranes via pore formation, leading to cell lysis. Metabolomic profiling further highlighted its ability to increase carbohydrate and amino acid metabolism, supporting cell growth and survival in acidic environments. Also, amino acid metabolism (elevated tryptophan, tyrosine, histidine) supports acid tolerance and immune modulation. Tryptophan metabolism produces indole derivatives, which are known to benefit gut health and immunity. Additionally, strain SK2-659 demonstrated strong tolerance to gastrointestinal conditions, high adhesion capacity to intestinal cells, and immunomodulatory effects, contributing to gut health. Safety assessments confirmed the absence of virulence factors, pathogenic traits, and antibiotic resistance genes, along with a lack of biogenic amine production, underscoring its suitability for food and health applications. These findings establish L. lactis SK2-659 as a promising probiotic candidate with potential industrial applications in functional foods, food preservation, and therapeutic products. Given its probiotic properties, antimicrobial activity, and safety profile, this strain is particularly suited for the food industry as a functional food ingredient and natural bio-preservative, as well as the healthcare and pharmaceutical industries for use in therapeutic probiotics and gut health formulations.
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Gelatin hydrolyzed by papaya latex enzymes as an alternative cryoprotectant for frozen raw Pacific white shrimp (Penaeus vannamei)
(2024-08-01)
Kittiphattanabawon, Phanat
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Sriket, Chodsana
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Nalinanon, Sitthipong
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Visessanguan, Wonnop
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Benjakul, Soottawat
The cryoprotective activity of gelatin hydrolyzed by papaya latex enzymes at degrees of hydrolysis (DH) of 5%, 10%, 15%, and 20% were determined. The thermal hysteresis of gelati hydrolysate increased as DH increased up to 15% (2.31–4.19 °C) and decreased at DH of 20% (2.94 °C). The gelatin hydrolysate with 15%DH (G15), showing the highest thermal hysteresis, was subjected to fractionation and cryoprotective effect study in Pacific white shrimp. The fractionated peptide with the highest thermal hysteresis (5.19 °C) had a molecular weight of 1.7–4.2 kDa. After the soaking, G15-treated shrimp showed significantly lower and higher weight gain (7.33%) than those treated with mixed phosphates (16.12%) and distilled water (0.82%), respectively. Mixed phosphates- and G15-treated shrimp had lower α-glucosidase (AG) and β-N-acetyl-glucosaminidase (NAG) activities in exudate and lower thawing loss than distilled water-treated shrimp. G15-and mixed phosphates-treated shrimp also reduced freezable water content and maintained Ca2+-ATPase activity and protein solubility more effectively than distilled water-treated shrimp. Therefore, gelatin hydrolysate could be a potential alternative cryoprotectant in frozen raw shrimp.
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Revisiting chloroplast genomic landscape and annotation towards comparative chloroplast genomes of Rhamnaceae
(2023-12-01)
Wanichthanarak, Kwanjeera
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Nookaew, Intawat
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Pasookhush, Phongthana
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Wongsurawat, Thidathip
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Jenjaroenpun, Piroon
Background: Massive parallel sequencing technologies have enabled the elucidation of plant phylogenetic relationships from chloroplast genomes at a high pace. These include members of the family Rhamnaceae. The current Rhamnaceae phylogenetic tree is from 13 out of 24 Rhamnaceae chloroplast genomes, and only one chloroplast genome of the genus Ventilago is available. Hence, the phylogenetic relationships in Rhamnaceae remain incomplete, and more representative species are needed. Results: The complete chloroplast genome of Ventilago harmandiana Pierre was outlined using a hybrid assembly of long- and short-read technologies. The accuracy and validity of the final genome were confirmed with PCR amplifications and investigation of coverage depth. Sanger sequencing was used to correct for differences in lengths and nucleotide bases between inverted repeats because of the homopolymers. The phylogenetic trees reconstructed using prevalent methods for phylogenetic inference were topologically similar. The clustering based on codon usage was congruent with the molecular phylogenetic tree. The groups of genera in each tribe were in accordance with tribal classification based on molecular markers. We resolved the phylogenetic relationships among six Hovenia species, three Rhamnus species, and two Ventilago species. Our reconstructed tree provides the most complete and reliable low-level taxonomy to date for the family Rhamnaceae. Similar to other higher plants, the RNA editing mostly resulted in converting serine to leucine. Besides, most genes were subjected to purifying selection. Annotation anomalies, including indel calling errors, unaligned open reading frames of the same gene, inconsistent prediction of intergenic regions, and misannotated genes, were identified in the published chloroplast genomes used in this study. These could be a result of the usual imperfections in computational tools, and/or existing errors in reference genomes. Importantly, these are points of concern with regards to utilizing published chloroplast genomes for comparative genomic analysis. Conclusions: In summary, we successfully demonstrated the use of comprehensive genomic data, including DNA and amino acid sequences, to build a reliable and high-resolution phylogenetic tree for the family Rhamnaceae. Additionally, our study indicates that the revision of genome annotation before comparative genomic analyses is necessary to prevent the propagation of errors and complications in downstream analysis and interpretation.
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Functional genome analysis and anti-Helicobacter pylori activity of a novel bacteriocinogenic Lactococcus sp. NH2-7C from Thai fermented pork (Nham)
(2023-12-01)
Kingkaew, Engkarat
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Woraprayote, Weerapong
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Booncharoen, Auttaporn
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Niwasabutra, Kanidta
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Janyaphisan, Thitiphorn
Helicobacter pylori, linked to gastric diseases, is targeted for probiotic treatment through bacteriocin production. Bacteriocins have gained recognition for their non-toxic effects on host cells and their ability to combat a wide range of pathogens. This study aimed to taxonomically characterize and evaluate the safety and probiotic properties of the novel species of Lactococcus sp. NH2-7C isolated from fermented pork, as well as its bacteriocin NH2-7C, both in vitro and in silico. Comparative genotypic analysis revealed an average nucleotide identity of 94.96%, an average amino acid identity of 94.29%, and a digital DNA-DNA hybridization value of 63.80% when compared to Lactococcus lactis subsp. lactis JCM 5805T. These findings suggest that strain NH2-7C represents a novel species within the genus Lactococcus. In silico assessments confirmed the non-pathogenic nature of strain NH2-7C and the absence of genes associated with virulence and biogenic amine formation. Whole-genome analysis revealed the presence of the nisA gene responsible for nisin A production, indicating its potential as a beneficial compound with anti-Helicobacter pylori activity and non-toxic characteristics. Probiotic assessments indicated bile salt hydrolase and cholesterol assimilation activities, along with the modulation of interleukin-6 and tumour necrosis factor-α secretion. Strain NH2-7C demonstrated gastrointestinal tolerance and the ability to adhere to Caco-2 cells, affirming its safety and probiotic potential. Additionally, its ability to produce bacteriocins supports its suitability as a functional probiotic strain with therapeutic potential. However, further in vitro and in vivo investigations are crucial to ensure its safety and explore potential applications for Lactococcus sp. NH2-7C as a probiotic agent.
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Conjugated Autoxidizable Triene-Based (CAT and ApoCAT) Assays: Their Practical Application for Screening of Crude Plant Extracts with Antioxidant Functions in Relevant to Oil-in-Water Emulsions
(2019-01-01)
Phonsatta, Natthaporn
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Luangpituksa, Pairoj
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Figueroa-Espinoza, Maria Cruz
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Lecomte, Jérôme
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Durand, Erwann
Many previous reports suggested that conventional antioxidant assays could not forecast antioxidant performance of plant extracts, especially when it came to a real food system such as oil-in-water emulsion. In this study, antioxidant activities of aqueous and ethanolic plant extracts are investigated using multiple conventional assays (TPC, ABTS, FRAP, ORAC) and the oil-in-water (O/W) emulsion-based high throughput assays (the CAT and ApoCAT) in a comparison with an autoxidative O/W emulsion model monitored by formation of lipid hydroperoxide and TBARS values. Results suggest that only the ApoCAT assay is able to forecast the antioxidative performances of the extracts in O/W emulsions, regardless of the differences in extraction solvents, while the CAT assay can explain only the performance of ethanolic plant extracts in O/W emulsions. According to untargeted metabolite analysis, the antioxidants activities of plant extracts might be strongly influenced by extraction solvents. As a result, not only is the quantity of particular metabolites impacted, but also the whole metabolite (antioxidant) profiles of the extracts are modified. In addition, this study demonstrates that both the chemical reaction scheme and physical-state of a model are important parameters for designing a better antioxidant assay in the future. Practical Applications: The ApoCAT assay would be a more practical method for screening antioxidant compounds and/or crude plant extracts than conventional antioxidant assays and the original CAT assay. The antioxidant capacity of plant extracts obtaining from the ApoCAT can be used to represent their antioxidant performances in food emulsions regardless of their extraction solvents. Several crude plant extracts using different solvents are used to validate the performance of the CAT and the ApoCAT assays in comparisons with multiple conventional antioxidant assays. These assays differ from each other in terms of scheme of reaction and physical state of the test. In addition, oil-in-water (O/W) emulsion model is used as a reference method representing the antioxidant activities in food matrices. Principal component analysis (PCA) is performed to better understand the relationship among all assays. The results suggest that only the ApoCAT assay exhibits a close relationship with O/W emulsion model in both crude ethanolic and aqueous plant extracts. Furthermore, putative metabolite profiles of crude plant extracts are identified using LC-orbitrap MS/MS.
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Hydrolysed collagen from Lates calcarifer skin: its acute toxicity and impact on cell proliferation and collagen production of fibroblasts
(2018-08-01)
Benjakul, Soottawat
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Karnjanapratum, Supatra
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Visessanguan, Wonnop
Acute toxicity in Wistar rat and the impact of hydrolysed collagen (HC) from seabass skin on in vitro cell proliferation and collagen production were studied using L929 fibroblasts. HC was rich in glycine (326 residue/1000 residue) and imino acids (196 residue/1000 residue). MALDI mass spectrum of HC showed several low MW peptides with MW range of 1050–1330 Da as the major components. Based on acute oral cytotoxicity test in Wistar rat, HC was considered as safe with LD<inf>50</inf> value higher than 5000 mg kg−1 body. HC could promote L929 cell growth, especially when used in combination with vitamin C (VitC) at a ratio of 2:1. HC/VitC (2:1) mixture also exhibited the higher enhancement effect on collagen production of L929 cells, compared with HC or VitC alone. Thus, HC could be a promising candidate for biological applications, especially in combination with VitC, as nutraceuticals for skin care.
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Comparison of Antioxidant Evaluation Assays for Investigating Antioxidative Activity of Gallic Acid and Its Alkyl Esters in Different Food Matrices
(2017-08-30)
Phonsatta, Natthaporn
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Deetae, Pawinee
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Luangpituksa, Pairoj
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Grajeda-Iglesias, Claudia
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Figueroa-Espinoza, Maria Cruz
The addition of antioxidants is one of the strategies to inhibit lipid oxidation, a major cause of lipid deterioration in foods leading to rancidity development and nutritional losses. However, several studies have been reported that conventional antioxidant assays, e.g., TPC, ABTS, FRAP, and ORAC could not predict antioxidant performance in several foods. This study aimed to investigate the performance of two recently developed assays, e.g., the conjugated autoxidizable triene (CAT) and the apolar radical-initiated conjugated autoxidizable triene (ApoCAT) assays to predict the antioxidant effectiveness of gallic acid and its esters in selected food models in comparison with the conventional antioxidant assays. The results indicated that the polarities of the antioxidants have a strong impact on antioxidant activities. In addition, different oxidant locations demonstrated by the CAT and ApoCAT assays influenced the overall antioxidant performances of the antioxidants with different polarities. To validate the predictability of the assays, the antioxidative performance of gallic acid and its alkyl esters was investigated in oil-in-water (O/W) emulsions, bulk soybean oils, and roasted peanuts as the lipid food models. The results showed that only the ApoCAT assay could be able to predict the antioxidative performances in O/W emulsions regardless of the antioxidant polarities. This study demonstrated that the relevance of antioxidant assays to food models was strongly dependent on physical similarities between the tested assays and the food structure matrices.
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Bacteriocins from lactic acid bacteria and their applications in meat and meat products
(2016-10-01)
Woraprayote, Weerapong
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Malila, Yuwares
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Sorapukdee, Supaluk
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Swetwiwathana, Adisorn
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Benjakul, Soottawat
Meat and meat products have always been an important part of human diet, and contain valuable nutrients for growth and health. Nevertheless, they are perishable and susceptible to microbial contamination, leading to an increased health risk for consumers as well as to the economic loss in meat industry. The utilization of bacteriocins produced by lactic acid bacteria (LAB) as a natural preservative has received a considerable attention. Inoculation of bacteriocin-producing LAB cell as starter or protective cultures is suitable for fermented meats, whilst the direct addition of bacteriocin as food additive is more preferable when live cells of LAB could not produce bacteriocin in the real meat system. The incorporation of bacteriocins in packaging is another way to improve meat safety to avoid direct addition of bacteriocin to meat. Utilization of bacteriocins can effectively contribute to food safety, especially when integrated into hurdle concepts. In this review, LAB bacteriocins and their applications in meat and meat products are revisited. The molecular structure and characteristics of bacteriocins recently discovered, as well as exemplary properties are also discussed.
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Potential of bacteriocin-producing lactic acid bacteria for safety improvements of traditional Thai fermented meat and human health
(2015-11-01)
Swetwiwathana, Adisorn
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Visessanguan, Wonnop
Lactic acid bacteria (LAB) are very important in converting of agricultural products into safe, delicious and shelf stable foods for human consumption. The preservative activity of LAB in foods is mainly attributed to the production of anti-microbial metabolites such as organic acids and bacteriocins which enables them to grow and control the growth of pathogens and spoilage microorganisms. Besides ensuring safety, bacteriocin-producing LAB with their probiotic potentials could also be emerging as a means to develop functional meat products with desirable health benefits. Nevertheless, to be qualified as a candidate probiotic culture, other prerequisite probiotic properties of bacteriocin-producing LAB have to be assessed according to regulatory guidelines for probiotics. Nham is an indigenous fermented sausage of Thailand that has gained popularity and acceptance among Thais. Since Nham is made from raw meat and is usually consumed without cooking, risks due to undesirable microorganisms such as Salmonella spp., Staphylococcus aureus, and Listeria monocytogenes, are frequently observed. With an ultimate goal to produce safer and healthier product, our research attempts on the development of a variety of new Nham products are discussed.

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