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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
    ;
    Chotphrethipong, Lalita
    ;
    Benjakul, Soottawat
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    Hutamekalin, Pilaiwanwadee
    ;
    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
    ;
    ; ;
    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
    ;
    Sinthusamran, Sirima
    ;
    Benjakul, Soottawat
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    Senphan, Theeraphol
    ;
    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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    Impact of extraction condition on the yield and molecular characteristics of collagen from Asian bullfrog (Rana tigerina) skin
    (2022-06-01)
    Indriani, Sylvia
    ;
    Benjakul, Soottawat
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    Kishimura, Hideki
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    Karnjanapratum, Supatra
    ;
    The present study is the first report on collagen extraction as acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC), which could maximize the exploitation of Asian bullfrog (Rana tigerina) skin as an alternative collagen material in food and pharmaceutical fields. Extraction methods for ASC and PSC from Asian bullfrog skin were developed using different concentrations of acetic acid (0.50, 0.75, and 1.00 M) and pepsin (1.00, 2.00, and 3.00% [w/w]). PSCs exhibited a higher extraction yield (22.59%–28.30%) and recovery (22.48%–31.12%) than ASCs. An electrophoretic study revealed that collagens possessed type I collagen. ASC with 0.75 M acetic acid (ASC-0.75) and PSC with 3.00% (w/w) pepsin (PSC-3.00) were selected and analysed on their solubility, molecular characteristics, and amino acid composition. A better solubility with a higher isoelectric point was observed for PSC-3.00 than ASC-0.75. Fourier transform infrared- and circular dichroism-spectra revealed a high-interacted compact structure with a triple helical-structure of the collagen peptides, respectively. A similar amino acid profile with a high imino acid content was observed. Therefore, collagens from Asian bullfrog skin were successfully extracted, where the optimized conditions provided a high yield with less adverse effects on collagen properties and molecular characteristics.
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    Shelf-life prediction of micro-encapsulated shrimp oil in different packages using empirical models
    (2020-01-01) ;
    Benjakul, Soottawat
    ;
    Pisuchpen, Supachai
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    Kaewthong, Pensiri
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    Micro-encapsulated shrimp oil (MSO) can be used to fortify food products and may be especially useful in the development of new functional foods. The shelf-life of packaged MSO was studied and sorption isotherms were examined. Moisture sorption isotherms and moisture sorption kinetics of MSO were evaluated at 30°C for water activity (a<inf>w</inf>) values ranging from 0.113 to 0.923 by a static gravimetric method. Empirical models were determined to predict the experimental data. The initial stage of moisture sorption by the MSO was relatively rapid and it decreased with time. All the sorption curves were found to be type III. The most suitable model for predicting the moisture sorption isotherm of MSO was the GAB model because it had the lowest percentage root mean square error (RMSE). The shelf-life of MSO packaged in polypropylene (PP), Nylon/linear low density polyethylene (Nylon/LLDPE) and metalized polyethylene terephthalate (metalized PET) pouches stored at 30°C and either 75% or 80% relative humidity (RH) was predicted by the GAB equation and the longest shelf-life of MSO (507 days in 80% RH and 725 days in 75% RH) was found in metalized PET packages. The empirical models can be useful for predicting the shelf-life of MSO.
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    Development of Yellow Discoloration in Sawai (Pangasianodon hypophthalmus) Muscle due to Lipid Oxidation
    (2023-12-01) ;
    Kittiphattanabawon, Phanat
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    Patil, Umesh
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    Benjakul, Soottawat
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    Senphan, Theeraphol
    In this study, we investigated the impact of lipid oxidation on the discoloration of Sawai (Pangasianodon hypophthalmus) lipids and proteins. Sawai microsomes, liposomes, and salt-soluble myofibrillar proteins were prepared and subjected to lipid oxidation process. The results revealed that the levels of thiobarbituric acid-reactive substances, yellowness (as indicated by b* values), and pyrrole compounds increased when Sawai liposomes and microsomes were oxidized using iron and ascorbate. Meanwhile, the levels of free amines decreased, particularly as the iron content (25∼100 μM) and incubation time (0∼20 h) increased. The impact of oxidized liposomes at different levels (1, 2, and 5%) on the salt-soluble Sawai myofibrillar proteins was also evaluated. The findings revealed that lipid oxidation products reduced the sulfhydryl content and increased the surface hydrophobicity and carbonyl content of the salt-soluble Sawai myofibrillar proteins. These results imply that the formation of yellow discoloration in Sawai muscle could be due to nonenzymatic browning reactions occurring between lipid oxidation products and amines in the muscle protein.
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    Impact of different ultrasound-assisted processes for preparation of collagen hydrolysates from Asian bullfrog skin on characteristics and antioxidative properties
    (2022-09-01)
    Indriani, Sylvia
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    Sae-leaw, Thanasak
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    Benjakul, Soottawat
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    Hong Quan, Tran
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    Karnjanapratum, Supatra
    This study focused on impact of ultrasound-assisted process (UAP) at pre-treatment (UP) and simultaneous treatment (US) during papain hydrolysis for preparing collagen hydrolysate (CH) from Asian bullfrog skin. Ultrasonication times were varied (10, 20, 30 min), and CH prepared using papain hydrolysis without UAP was used as control. Different UAPs provided CH with various hydroxyproline contents, α-amino group contents, surface hydrophobicities, and antioxidative activities. UP at 20 min (UP-20) and US at 30 min (US-30) provided highly antioxidative CHs, which were selected for further studies on their Oxygen reactive absorbance capacity (ORAC) and molecular characteristics. CHs from UP-20 and US-30 had higher ORAC than that of control group (p ≤ 0.05). Slight difference in amino acid composition was detected between samples. Based on these results, molecular characteristic styles, molecular weight profile, antioxidative peptide content, and secondary structure of each sample were obtained. These results indicate that UP and US used varied enzymatic hydrolysis patterns and modified molecular conformation of CH, resulting in enhanced antioxidative activity. Therefore, different UAPs as UP and US could be effectively used in preparation of CH using papain hydrolysis from Asian bullfrog skin, which could improve production process efficiency by enhancing their bioactivity, particularly antioxidative activity.
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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
    ;
    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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    Ultrasound-assisted extraction of collagen from clown featherback (Chitala ornata) skin: yield and molecular characteristics
    (2021-01-30)
    Petcharat, Tanyamon
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    Benjakul, Soottawat
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    Karnjanapratum, Supatra
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    BACKGROUND: Clown featherback (Chitala ornata) skin, a by-product from the filleting process line, could serve as a good aquatic collagenous source. Nevertheless, the typical collagen extraction method is a time-consuming process providing a relatively low yield. Ultrasound had been reported to be an alternative technique for enhancing the extraction efficiency of several compounds, although the harsh conditions of ultrasound could affect their physicochemical and molecular characteristics. Thus, the application of ultrasonication under appropriate conditions could comprise a promising means for improving the extraction efficiency of collagen from clown featherback skin. RESULTS: Ultrasonication using different amplitudes (20–80%) and times (10–30 min) was implemented during extraction. An ultrasound-assisted process (UAP) was able to increase the yield of collagen (P ˂ 0.05) and could also result in a collagen purity decrease as evaluated by hydroxyproline content. There was no dramatic change in the solubility of resulting collagens. UAP induced protein degradation, particularly with an increasing amplitude and time, where slight changes in the isoelectric point value of collagen were observed. UAP had no adverse effect on molecular structure, where a triple-helical structure was still retained when an 80% amplitude was employed for 10 min (UAP-80/10-C). The amino acid composition of UAP-80/10-C reconfirmed the unique characteristic of collagen containing imino acid. CONCLUSION: An UAP under appropriate conditions could be used to improve the extraction yield with minimal effects on the molecular integrity of the resulting collagen. In addition, fish skin waste from the cutting process line, particularly clown featherback skin, could be exploited as a value-added product, comprising fish skin collagen. © 2020 Society of Chemical Industry.