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    Pulsed electric field-assisted extraction of Djenkol (Archidendron pauciflorum) peel: Characterization, suppression of intracellular ROS generation and inflammatory cytokines in LPS-activated RAW264.7 macrophage cells
    (2024-06-01)
    Sinthusamran, Sirima
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    Chotphrethipong, Lalita
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    Benjakul, Soottawat
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    Hutamekalin, Pilaiwanwadee
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    Champoochana, Nidanut
    Djenkol (Archidendron pauciflorum) peel is a source of phenolics with an antioxidative activity. Nevertheless, the high extraction efficacy of the target compounds is crucial for the isolation of such compounds. Pulsed electric field (PEF)-assisted extraction is the means that increase the efficacy of extraction via an electroporation mechanism. This research aimed to study the extraction of phenolics with antioxidative activities using the PEF and to investigate the anti-inflammation and antioxidative activities of the Djenkol peel extract (DPE) in RAW264.7 macrophage cells. The PEF at different electric field strengths (E) (4.5 and 6 kV/cm) and times (180, 360, and 540 ms) was implemented to extract the phenolic compounds. The PEF with the E level at 6 kV/cm for 540 ms provided the highest yield, the total phenolic content, and antioxidative activities, compared to other conditions (P < 0.05). The dominant compounds in the DPE were the gallic acid and catechin. When the RAW264.7 cells were treated with the DPE at different levels (0.125, 0.25, 0.5, 0.75 and 1 μg/mL), the DPE at 0.125 and 0.25 μg/mL had no cytotoxicity, compared to the control (P < 0.05). With adding lipopolysaccharide (LPS), the DPE-treated cells at 0.125 μg/mL showed the similar cell viability to that of the control (P < 0.05). Additionally, the use of the DPE, particularly at 0.125 and 0.25 μg/mL, could reduce the TNF-α and IL-6 levels and reactive oxygen species (ROS) generation in the LPS-activated cells. Thus, the DPE could be used as a functional ingredient in food.
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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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    ; ;
    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 Ca<sup>2+</sup>-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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    Development of djenkol peel extract-loaded liposome as functional food ingredients: Physicochemical characteristic, antioxidant activities, and cytotoxicity
    (2025-06-01)
    Chotphruethipong, Lalita
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    Sinthusamran, Sirima
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    Benjakul, Soottawat
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    Senphan, Theeraphol
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    Djenkol peel is an agricultural by-product containing valuable bioactive compounds. The direct use of its extract faces challenges related to stability and sensory acceptance. This study aimed to investigate the effect of djenkol peel extract (DPE) on the encapsulation efficiency (EE) using liposomes. The physicochemical properties, antioxidant activities, and cytotoxicity of the obtained liposomes were evaluated. DPE (0, 0.25, 0.50, 0.75, and 1.00 %, w/v) was encapsulated using phospholipid/cholesterol as the wall material. The highest EE (66.69 %) was observed in the liposomes loaded with 0.25 % (w/v) DPE (LE-0.25DPE) (P < 0.05). All DPE-loaded liposomes displayed nanometer-scale particle sizes (78.94–135.60 nm) and negative zeta potentials (−52.60 to −57.50 mV). Encapsulation of DPE in liposomes can mask its undesirable color. Fourier-transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), and differential scanning calorimetry (DSC) revealed changes in the physicochemical and thermal properties of LE-0.25DPE. Additionally, LE-0.25DPE showed increased DPPH radical-scavenging activity (DPPH-RSA) and ferric reducing antioxidant power (FRAP) in a dose-dependent manner (250–2000 μg/mL). In vitro Caco-2 cell viability test, LE-0.25DPE at 0.1–1 μg/mL exhibited no cytotoxicity compared to the control (P < 0.05). Thus, DPE-loaded liposomes demonstrate potential as a functional food ingredient with high bioactivity.
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    Elucidating the physicochemical and structural properties of Ganoderma lucidum spores: Comparative analysis of various disruption techniques
    (2024-04-15) ;
    Kuimalee, Surasak
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    Yarnpakdee, Suthasinee
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    Benjakul, Soottawat
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    Sriket, Pornpimol
    This study evaluated Ganoderma lucidum G2 spores (GLS) using various processing techniques: vibrating milling, ball milling, and autoclaving. Broken GLS via vibrating and ball milling showed higher lipid content (22.89% and 22.50%) but lower L*, a*, and b* values compared to intact GLS. Vibrating milling achieved the highest sporoderm-broken rate (98.27%). Ball milling resulted in the highest lipid oxidation. Vibrating milling yielded the highest bioactive compounds, and both methods increased DPPH-RSA values. Differential scanning calorimetry indicated varying T<inf>max</inf> values: non-broken GLS (106.56 °C), vibrating milling (92.77 °C), ball milling (97.58 °C), and autoclaving (109.35 °C). FT-IR spectra showed triterpenes, polysaccharides, and fatty acids. X-ray diffraction and Fourier transform infrared spectroscopy revealed increased crystallinity with vibrating milling (37.12% to 47.99%). Both milling methods altered GLS structure to a disintegrated, pancake-like form. Scanning electron microscopy with energy-dispersive X-ray spectroscopy indicated significant carbon and oxygen concentrations. This study provided crucial insights for utilizing GLS.
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    Optimization of salt washing protocols for surimi production from invasive blackchin tilapia (Sarotherodon melanotheron): Molecular and functional properties
    (2025-10-01)
    Senphan, Theeraphol
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    Mungmueang, Natthapong
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    Hoque, Md Sazedul
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    Benjakul, Soottawat
    This study presents the first systematic molecular characterization and protein chemistry optimization for surimi processing from invasive blackchin tilapia (Sarotherodon melanotheron), introducing novel salt washing protocols. Ten treatments were evaluated including conventional washing, single/multiple salt washing (0.3–0.9 % NaCl), and pH-shifting methods. Molecular analyses (SDS-PAGE, ATR-FTIR and SEM) revealed myofibrillar protein preservation and distinct structural modifications. The optimized 0.6 % NaCl protocol achieved selective heme protein removal while preserving myofibrillar integrity, yielding superior gel chemistry (whiteness: 76.47 vs. 71.04) and highest sensory acceptance (7.33 vs. 6.40). pH-shifting enhanced protein recovery (68.59 %) but caused molecular aggregation and network disruption, reducing gel strength by 82 %. Molecular analysis demonstrated that salt washing selectively removes chromoproteins while preserving essential gel-forming proteins, whereas pH-shifting caused protein aggregation. SEM confirmed dense protein matrices in salt-washed samples versus fragmented networks in pH-treated surimi. This chemistry-guided approach transforms invasive species into functional protein ingredients.
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    Alpha-amylase inhibitory activity of collagen hydrolysate from Asian bullfrog skin and its application in dark chocolate
    (2024-01-01)
    Indriani, Sylvia
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    Benjakul, Soottawat
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    Sitanggang, Azis Boing
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    Karnjanapratum, Supatra
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    The present study aimed to investigate the in-vitro antidiabetic activity of collagen hydrolysate (CH) from Asian bullfrog skin and its application as a bioactive compound in dark chocolate bars. Papain (3% [w/w]) was used to obtain CH with a simultaneous conjunction of ultrasound. CH possessed α-amylase inhibitory activity with an IC<inf>50</inf> value of 3.60 mg/mL, while results from inhibition reaction kinetics revealed that CH inhibited α-amylase in a non-competitive mode. Different concentrations (0.5, 1.0, and 2.0% [w/w]) of CH were then fortified into dark chocolate bars, and the physicochemical characteristics, breaking strength, and rheological behaviour of the resulting chocolate were investigated and compared with a control (without CH addition). A higher level of CH significantly enhanced antidiabetic activity against α-amylase (p ≤ 0.05) in chocolate bar. Chocolate bar with 2% CH complied with the nutritional claim requirement for ‘source of protein’ with lower fat content and energy value, while sensory evaluation revealed good organoleptic properties.
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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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    ; ;
    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 Ca<sup>2+</sup>-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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    Extraction techniques, structural features, and functional properties of collagenous derivatives from unconventional animal sources: a review
    (2026-12-01)
    Indriani, Sylvia
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    Petcharat, Tanyamon
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    Andriani, Cynthia
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    Benjakul, Soottawat
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    Collagenous derivatives (collagen, gelatin, and collagenous hydrolysate (CH)) are extensively used across the food, biomedical, and pharmaceutical industries. Traditionally, these have been sourced from porcine, bovine, and fish due to their ready availability and biocompatibility. However, conventional collagenous derivatives face ongoing challenges regarding sustainability, resource intensity, and socio-cultural perceptions. This has led to the exploration of alternative collagenous derivatives from unconventional sources, with a primary focus on evaluating their potential for yields, extractability, and functional properties, all of which are fundamental for future scale-up and alternative applications. This review summarizes alternative collagenous derivatives from unconventional animals, including amphibians, mollusks, echinoderms, insects, unconventional fish and byproducts, and reptiles. Their structures, extraction techniques, functional properties, and potential applications are comprehensively summarized, showcasing their ability to complement or even surpass conventional sources in specific uses. Additionally, the challenges and prospects for industrial application, emphasizing the sustainability of meeting growing collagen demand and encouraging further research into these promising alternative sources, were discussed. Unconventional collagenous derivatives demonstrate excellent and unique characteristics as alternatives to conventional ones. Type I collagen from amphibians, reptiles, and mollusks had superior thermal stability. Unconventional gelatin and CH also possess various bio-functionalities that can enhance their potential applications. The relatively low extraction yield could be addressed by increasing the concentration of chemicals or extraction time and incorporating green technology without causing an adverse impact on the quality. These findings indicate the potential applications of unconventional collagenous derivatives as food ingredients and supplements.