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    Enhancing Biomass and Lipid Production in Messastrum gracile Using Inorganic Carbon Substrates and Alternative Solvents for Lipid Extraction
    (2025-03-01)
    Pan-utai, Wanida
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    Inrung, Wilasinee
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    Thurakit, Theera
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    Srinophakun, Penjit
    Microalgae show promise as a biomass and bioproduct for applications in various industries. The cultivation of microalgae plays a crucial role in optimizing biomass yield and bioproduct accumulation. The provision of inorganic carbon substrates substantially enhances microalgal growth and lipid biosynthesis, resulting in marked increases in the production of biofuels and other bioproducts. This study examined biomass and lipid accumulation in Messastrum gracile IFRPD 1061 under inorganic stress conditions, previously unreported. M. gracile IFRPD 1061 was subjected to varying conditions of inorganic carbon substrates, 1–3 g·L<sup>−1</sup> sodium carbonate and bicarbonate concentration, to enhance biomass and lipid accumulation. Optimal productivity levels were observed with sodium bicarbonate addition of 3 g·L<sup>−1</sup> and 1 g·L<sup>−1</sup> for biomass and lipids, resulting in productivities of 392.64 and 53.57 mg·L<sup>−1</sup>·d<sup>−1</sup>, respectively. Results underlined the effectiveness of sodium carbonate and bicarbonate as inorganic carbon sources for stimulating microalgal growth and enhancing the production of high-value products. The extraction of lipids from freeze-dried biomass of M. gracile IFRPD 1061 demonstrated optimal yield using methanol/hexane solvents compared with the control experiments. Lipid extraction yields using methanol/hexane were 42.18% and 46.81% from oven-dried and freeze-dried biomass, respectively. Lipids extracted from oven-dried M. gracile IFRPD 1061 using methanol/hexane/chloroform solvents indicated the potential of methanol/hexane as a solvent for lipid extraction from dry microalgal biomass using an ultrasonic-assisted technique. This study contributes valuable insights into maximizing biofuel and bioproduct production from microalgae, highlighting A. gracilis as a promising candidate for industrial applications.
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    Development of a synbiotic longan beverage: a nutritious and functional blend
    (2025-02-01)
    Sakuntasri, K.
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    Massa, S.
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    Krusong, W.
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    Charoenrat, T.
    Synbiotic foods, which synergistically combine probiotics and prebiotics, have garnered attention for their potential health benefits. In line with this trend, this study aimed to develop a synbiotic beverage using longan fruit as the base and fructo-oligosaccharides (FOS) as the prebiotic source. Two strains of Lactobacillus probiotics, Lactobacillus casei 431 and Lactobacillus plantarum KM001, were used to ferment the longan juice with and without FOS. Total soluble solids, reducing sugar, pH, total acid production, and viable cell count of Lactobacillus microorganisms were evaluated to determine the optimal fermentation duration. The findings indicated that fermenting the juice for 12 hrs at 37°C aligned with the criteria established by the FAO/WHO for probiotic foods. Furthermore, compared to L. casei 431, the longan juice containing FOS fermented with L. plantarum KM001 showed a better survival rate in a simulated gastrointestinal environment. Additionally, the microorganism count in this beverage remained relatively constant throughout its 28-days shelf life at 4°C. These findings underscore the synbiotic longan beverage as a functional drink that combines viable probiotics with prebiotic FOS.
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    Valorizing Red Seaweed Spent Biomass into Reducing Sugars for β-Carotene Production by Rhodotorula paludigena
    (2026-05-01)
    Kongsinkaew, Chatchol
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    Tangsattayatithan, Chutipol
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    Chittapun, Supenya
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    Phiphatbunyabhorn, Parivat
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    Laemthong, Tunyaboon
    Seaweed bioactive extraction generates de-extracted residual solids that remain carbohydrate-rich but are often underutilized. This study developed an integrated valorization route for Gracilaria fisheri spent biomass to produce fermentable sugars for β-carotene production by Rhodotorula paludigena CM33. Reducing sugar production was optimized using response surface methodology (Box–Behnken design) by varying reaction time, sulfuric acid concentration, and biomass loading at 90 °C. The predicted optimum (47.39 min, 2.50% (w/v) H<inf>2</inf>SO<inf>4</inf>, and 7.13% (w/v) biomass) yielded 22.41 g/L reducing sugars and was validated experimentally at 22.22 ± 0.19 g/L, indicating that the model reliably predicted reducing sugar production. The optimized condition was scaled up in a 22 L bioreactor with sequential acid hydrolysis followed by enzyme-assisted hydrolysis, increasing reducing sugars from ~30 to ~40 g/L. FTIR and SEM analyses indicated progressive modification of the carbohydrate matrix across processing stages. Batch cultivation of R. paludigena on the hydrolysate showed that ammonium sulfate supplementation significantly increased biomass, whereas β-carotene titers were not significantly different. Repeated-batch operation on non-supplemented hydrolysate sustained production over four cycles with β-carotene titers of 13.75–17.27 mg/L, demonstrating the operational feasibility of the hydrolysate-based system. Overall, this work demonstrates a practical seaweed biorefinery approach to upgrade G. fisheri spent biomass into sugars and carotenoid-rich yeast biomass.
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    Thermal degradation kinetics and purification of C-phycocyanin from thermophilic and mesophilic cyanobacteria
    (2025-02-01)
    Chittapun, Supenya
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    Suwanmanee, Kattiya
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    Kongsinkaew, Chatchol
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    Chisti, Yusuf
    The natural blue colorant C-phycocyanin (C-PC) has many potential applications but its poor heat stability limits its commercial use. This study compares the production and thermal stability of C-PC from two cyanobacteria: the thermophilic Thermosynechococcus sp. TUBT-T01 and the mesophilic Synechococcus cedrorum TISTR8589. Thermosynechococcus sp. produced nearly 1.9-fold more C-PC than S. cedrorum. Batch adsorption using a chromatographic cationic ion exchange resin (Streamline Direct HST1) was used to effectively purify the C-PC. The equilibrium adsorption capacity (Q<inf>eq</inf>) of the resin for C-PC was the highest at pH 5. At this pH, the Q<inf>eq</inf> for the thermophilic C-PC was 5.5 ± 0.1 mg mL⁻¹, whereas for the mesophilic C-PC it was 1.5 ± 0.2 mg mL⁻¹. Purification increased the concentration of the thermophilic C-PC by 5.9-fold, and that of mesophilic C-PC by 4.2-fold. The purity ratios of the final products from the two cyanobacteria were similar at ∼2.2. At 60 °C and pH 7, the C-PC of Thermosynechococcus sp. had ∼12-times longer half-life than the mesophilic C-PC; however, the productivity of the thermophilic C-PC was comparatively low because of a low biomass productivity of Thermosynechococcus sp.
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    Fed-batch strategies for growing β-carotene enriched yeast Rhodotorula paludigena using untreated molasses
    (2025-11-01)
    Kongsinkaew, Chatchol
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    Chisti, Yusuf
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    Ketudat-Cairns, Mariena
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    Chittapun, Supenya
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    The β-carotene-rich yeast Rhodotorula paludigena CM33 was grown in a 22-L bioreactor using sugarcane molasses in combination with several distinct fed-batch strategies: DO (dissolved oxygen)-stat; stepwise feeding; and hybrid feeding (an initial exponential feeding phase followed by a DO-stat operation). The impacts of feeding strategies on biomass production, carotenoid production and nutrient conversion efficiencies were evaluated. The stepwise feeding proved most effective, achieving: a final dry biomass concentration of 73.1 ± 2.7 g L<sup>−1</sup>; a β-carotene titer of 181.9 ± 1.0 mg L<sup>−1</sup>; a biomass volumetric productivity of 1.33 ± 0.05 g L<sup>−1</sup> h<sup>−1</sup>; and β-carotene productivity of 3.31 ± 0.02 mg L<sup>−1</sup> h<sup>−1</sup>. However, with this feeding strategy the sugar and nitrogen conversion efficiencies were the lowest: the β-carotene yield on total sugar was 0.92 ± 0.00 mg g<sup>−1</sup> and its yield on nitrogen was 5.47 ± 0.06 mg g<sup>−1</sup>. In contrast, the DO-stat operation was the most resource efficient, providing a biomass yield on sugar of 0.47 ± 0.02 g g<sup>−1</sup>, a biomass yield on nitrogen of 2.64 ± 0.05 g g<sup>−1</sup>, a β-carotene yield on sugar of 1.08 ± 0.04 mg g<sup>−1</sup>, and β-carotene yield on nitrogen of 6.08 ± 0.08 mg g<sup>−1</sup>. The hybrid strategy resulted in intermediate values of the various performance parameters. Raw sugarcane molasses proved to be excellent for high-cell-density culture of R. paludigena. A compromise between productivity and resource use efficiency may be required for commercial production of β-carotene especially if the substrate is expensive compared to the product.
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    Natural versus Saccharomyces boulardii self-induced anaerobic coffee fermentation: Effects on physicochemical properties and microbial ecology, and their influence on volatile profiles and sensory attributes across roast levels
    (2025-10-01)
    Pholtaisong, Jatuphol
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    Kongsinkaew, Chatchol
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    On-Mee, Thapanut
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    Chittapun, Supenya
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    This study investigates the effects of natural self-induced anaerobic fermentation (NSIAF) and Saccharomyces boulardii self-induced anaerobic fermentation (SSIAF) on Arabica coffee during wet processing. Over 24 h of fermentation, NSIAF exhibited greater microbial diversity, higher titratable acidity, and increased reducing sugar consumption, while SSIAF provided a more controlled microbial environment dominated by S. boulardii. Volatile compound analysis identified 207 compounds, with lighter roasts showing the greatest number of significantly different compounds between NSIAF and SSIAF treatments. Sensory evaluation revealed a higher cupping score for NSIAF at a light roast (82.08 ± 0.14) compared to SSIAF (81.58 ± 0.14), reflecting distinct flavor characteristics imparted by each fermentation process. Both methods achieved specialty coffee standards (≥80 points), highlighting the potential of NSIAF for complex and diverse profiles and the suitability of SSIAF for consistency and controlled fermentation.
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    High-cell-density culture for recombinant xylanase production and its application in hydrolysis of mild alkaline pretreated rice straw for xylooligosaccharide production
    (2026-08-01)
    Laemthong, Tunyaboon
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    Sukhumsirichart, Wasana
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    Chittapun, Supenya
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    Kongsinkaew, Chatchol
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    Satipattarn, Atikant
    Rice straw is an abundant lignocellulosic residue whose effective utilization requires coordinated enzyme performance and pretreatment conditions. In this study, recombinant xylanase production was integrated with mild alkaline pretreatment to enable efficient conversion of rice straw into xylooligosaccharides (XOS). Recombinant xylanase was produced by Escherichia coli using high-cell-density fed-batch cultivation, achieving a maximum activity of 207.95 U/mL in an enriched synthetic medium. Alkaline pretreatment was optimized using response surface methodology, identifying 6.75% (w/v) biomass loading, 193.17 mM NaOH, and 20 min as conditions that balanced solid recovery with sugar release during subsequent enzymatic hydrolysis. Hydrolysis of the pretreated rice straw yielded 6.13 mg/mL total XOS, mainly xylotriose (X3, 2.73 mg/mL; 44.56%), xylobiose (X2, 2.00 mg/mL; 32.62%), and xylotetraose (X4, 1.02 mg/mL; 16.57%), with only minor amounts of xylose and xylopentaose detected. The combined X2 and X3 fractions accounted for 77.18% of the total XOS. XOS with degrees of polymerization of 2–4 are recognized as effective prebiotics, particularly for beneficial intestinal bacteria such as Bifidobacterium spp. Overall, linking recombinant enzyme production with alkaline pretreatment improved conversion consistency and supported efficient XOS production from rice straw.