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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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Nalinanon, Sitthipong
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.
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Development of instant germinated black rice (Maejo rice 1A) milk beverage powder fortified with Cordyceps militaris protein isolate
(2026-08-01)
Senphan, Theeraphol
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Mungmueang, Natthapong
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Jaturapanyaskul, Siri
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Wangcharoen, Wiwat
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Daengprok, Wichittra
Development of instant germinated black rice (Maejo rice 1A) milk beverage powder fortified with Cordyceps militaris protein isolate was investigated. The base formula was optimized using mixture design, with Formula 8 (24% rice powder, 12% monk fruit sugar, 44% milk powder, 19.7% inulin and 0.3% Palsgaard® RecMilk) selected based on sensory evaluation. The beverage was then fortified with protein isolate at 0%, 3%, 5%, and 7% (w/w). Physicochemical properties, antioxidant activities, and sensory characteristics were evaluated. Protein fortification influenced color, water activity, viscosity, and nutritional composition, while enhancing phenolic-related compounds and antioxidant potential, indicating improved functional properties of the beverage powder. The 7% protein-fortified sample exhibited the highest antioxidant activity across all assays while maintaining acceptable thermal stability. Sensory evaluation showed that 5% protein fortification achieved optimal consumer acceptance (overall score 7.65), outperforming commercial products. The study demonstrates successful development of a functional beverage powder with enhanced nutritional and antioxidant properties through protein fortification.
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Enhanced calcium solubility of bio-calcium powder from blackchin tilapia (Sarotherodon melanotheron) bone: Influence of alkaline pretreatment conditions
(2026-07-01)
Senphan, Theeraphol
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Mungmueang, Natthapong
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Sazedul Hoque, Md
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Sriket, Chodsana
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Benjakul, Soottawat
Bio-calcium powder was prepared from blackchin tilapia ( Sarotherodon melanotheron ) bones, an invasive species by-product from surimi production in Thailand, to evaluate the effects of alkaline pretreatment on bio-calcium properties. Bones were pretreated with NaOH or KOH (1 and 2 M) at a bone-to-solution ratio of 1:10 (w/v) for 2 h, followed by ethanol washing, hydrogen peroxide bleaching, drying, and grinding to obtain bio-calcium powder. The yield on a wet weight basis was 26–28%, with water activity values of 0.33–0.34 measured at room temperature. Among all treatments, 1 M KOH gave the highest yield (28%), calcium content (26.13%), and lightness (L∗ = 88.5). Collagenous proteins and hydroxyapatite were identified by SDS-PAGE and ATR-FTIR, respectively. All bio-calcium samples showed four to five times higher in vitro calcium solubility after simulated gastrointestinal digestion (13.4-17.50%) than calcium carbonate (3.4%). The 2 M treatments exhibited the highest calcium solubility (17.2-17.50%). The 1 M KOH can be used for food fortification due to its high yield and lightness, whereas the 2 M treatments can serve as calcium supplements where higher calcium solubility is required. Blackchin tilapia bone is a potential source for bio-calcium production, with 1 M KOH suitable for food fortification and 2 M treatments more appropriate for applications requiring higher calcium solubility.
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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 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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Sequential extraction protocol for bio-calcium from saltwater crocodile (Crocodylus porosus) bone: Physicochemical and molecular studies
(2026-06-01)
Yarnpakdee, Suthasinee
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Senphan, Theeraphol
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Benjakul, Soottawat
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Sriket, Chodsana
Saltwater crocodile ( Crocodylus porosus ; SC) farming generates 45% bone waste, creating environmental challenges but also representing an underutilized source of calcium. This study presents the first comprehensive protocol for extracting bio-calcium from SC bone through sequential processing: autoclaving, alkaline soaking, ethanol extraction, and hydrogen peroxide bleaching. The extraction process yielded 16.94% bio-calcium with a high mineral density (26.37% Ca and 13.83% P) and an average particle size of 6.77 μm. XRD confirmed the preserved hydroxyapatite structure, while FTIR revealed the complete elimination of the organic matrix in the resulting bio-calcium. SEM-EDS demonstrated uniform elemental distribution with no detectable heavy metals. Remarkably, in vitro calcium bioavailability reached 15.24%, representing a 78% enhancement over fish bone bio-calcium and doubling synthetic supplement performance. Preserved collagen-derived amino acids facilitate calcium-peptide chelation, explaining the superior absorption. The developed valorization process converts aquaculture waste into high-value ingredients with potential applications in food and pharmaceuticals.
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Enhanced Stability and Bioavailability of Defatted Cricket Protein Hydrolysates Encapsulated in Alginate-Coated Liposomes
(2026-04-01)
Chotphruethipong, Lalita
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Benjakul, Soottawat
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Aluko, Rotimi E.
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Senphan, Theeraphol
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Hutamekalin, Pilaiwanwadee
The practical application of protein hydrolysates as functional food ingredients is frequently obstructed by their inherent structural instability. To circumvent this limitation, liposomal encapsulation has emerged as a sophisticated strategy to bolster the bioactivity and integrity of cricket-derived proteins. In this study, varying concentrations (1–4% w/v) of defatted cricket protein hydrolysate (DCPH) were integrated into vesicles composed of soy lecithin and cholesterol. The highest encapsulation efficiency (EE) was observed at a 2% DCPH loading capacity, yielding a significant result of 88.18% (p < 0.05). Subsequent coating with sodium alginate (SA) at 0.1–0.3% (w/v) resulted in an increase in particle size and a more pronounced negative surface charge. When maintained at 4 °C over a 24-day duration, the SA-coated liposome (SA-L-2%DCPH) exhibited superior stability compared to its uncoated (L-2%DCPH) counterpart. Also, the digest derived from the SA-L-2%DCPH exhibited significantly enhanced transepithelial permeability across the Caco-2 cell monolayer, indicated by the higher protein content and ABTS radical scavenging activity. Thus, sodium alginate-coated liposomes serve as a promising delivery system for encapsulating DCPH both during storage stability and in the gastrointestinal digestion system.
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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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Sriket, Chodsana
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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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Improving calcium bioavailability from fish bone waste: the role of sodium hydroxide in bio‑calcium extraction from tilapia (Oreochromis niloticus) bones
(2025-10-01)
Senphan, Theeraphol
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Mungmueang, Natthapong
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Choommongkol, Vachira
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Sriket, Chodsana
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Kishimura, Hideki
Calcium deficiency is a global health issue motivating the search for sustainable calcium sources from aquaculture waste. This study investigated sodium hydroxide (NaOH) concentration effects on bio‑calcium quality and bioavailability extracted from tilapia (Oreochromis niloticus) bones. Tilapia bones were treated with 0.5 M, 1 M, and 2 M NaOH solutions for 30 min compared to untreated controls. Increasing NaOH concentrations decreased yield while improving brightness, ash content, and calcium bioavailability. The 2 M NaOH treatment produced bio‑calcium with highest calcium bioavailability (8.57 %), surpassing both control (7.26 %) and commercial calcium carbonate (0.72 %) by 12-fold. Higher NaOH concentrations reduced moisture, protein, and fat contents while increasing hydroxyproline and decreasing lipid oxidation. SEM showed smoother surfaces with homogeneous pores in 2 M NaOH-treated samples. ATR-FTIR and EDS confirmed consistent hydroxyapatite structure. This optimization provides a sustainable approach for converting fish waste into high-bioavailability calcium supplements, supporting circular economy in aquaculture while addressing global calcium deficiency.
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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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Nalinanon, Sitthipong
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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Physicochemical properties and bioavailability of bio-calcium products from tilapia bone: A comparative study with synthetic hydroxyapatite
(2025-03-01)
Arkanit, Kornpaka
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Senphan, Theeraphol
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Issapap, Nursakich
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Mungmueang, Natthapong
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Sriket, Pornpimol
This study compared three calcium products derived from tilapia bones (fish bone powder, bio-calcium, and calcined bone) with synthetic hydroxyapatite as a reference material. The chemical compositions, physical properties, and calcium bioavailability were analyzed. The yield percentage decreased from 44.4 % in fish bone powder to 27.4 % in bio-calcium. The yield percentage then increased to 64.9 % in calcined bone. Synthetic hydroxyapatite showed 100 % yield. The ash content increased progressively from 68.5 % to 99.4 %. The calcium content increased from 18.9 % to 31 %, with Ca/P ratios improving from 3.3 to 1.7, approaching the ideal ratio of natural bone mineral (1.67). Heavy metals were not detected in any samples. All products exhibited high whiteness values (L∗ 91.8–95.0). Water activity values ranged from 0.33 to 0.50. The median particle sizes varied between 17.39 and 31.05 μm. Lipid oxidation decreased through the processing stages. Calcium bioavailability was the highest in synthetic hydroxyapatite (5.4 %). Bio-calcium showed 4.1 % bioavailability. The products demonstrated potential for food fortification and biomedical applications. The findings support the use of tilapia bones as a sustainable source of calcium production.

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