Xylanases, a subclass of glycoside hydrolases, catalyze the breakdown of xylan into xylan disaccharides, oligosaccharides, and xylose, playing a critical role in the degradation and utilization of hemicelluloses. However, extreme conditions hinder their broader industrial applications. In this study, a GH10 xylanase gene (referred to as taxyn10a) was sourced from Thermomonsospora amylolytica, cloned, and heterologously expressed in Escherichia coli BL21(DE3). The recombinant TaXyn10A was purified using Ni2+-affinity chromatography. Its optimal temperature and pH were determined to be 65°C and 5.6, respectively. The enzyme maintained over 80% of its activity between 55-70°C, with about 60% residual activity after 120 minutes at 60°C. Additionally, it preserved more than 80% of its activity in a pH range of 3.6-6.6. TaXyn10A exhibited over 60% activity in the presence of 1 or 10 mM ions (K+, Fe3+, Mg2+, Ca2+, Zn2+, Mn2+, Ni2+, Co2+, Pb2+, Cu2+), as well as 1% EDTA, PMSF, and DTT. Kinetic analysis with beechwood xylan yielded apparent Vmax, Km, and kcat values of 69.44 μmol min⁻¹ mg⁻¹, 24.53 mg mL⁻¹, and 53.35 s⁻¹, respectively. These findings demonstrate that TaXyn10A was active over moderately acidic pH and elevated-temperature ranges and tolerates various metal ions and chemical additives, highlighting its potential for feed processing and lignocellulosic biomass conversion.
This study investigates the degradation of Methyl Orange (MO) using CuSnO3 (CSO) modified with monoethanolamine (MEA) as a chelating and structure-directing additive and synthesized by the sol-gel method. The effects of MEA concentration on the structural, morphological, optical, and photocatalytic properties of CSO were systematically examined. X-ray diffraction (XRD) confirmed that all samples exhibited the cubic spinel CuSnO3 phase, and MEA enhanced crystallinity and improved structural order. Scanning electron microscopy (SEM) showed that MEA altered particle morphology from irregular, strongly agglomerated grains to more uniform spherical and octahedral structures. Ultraviolet diffuse reflectance spectroscopy (UV-DRS) and Tauc plot analysis indicated optical band gap values of 2.4, 3.13, 2.79, and 2.65 eV for the unmodified, 1:1, 1:2, and 2:1 MEA samples, respectively. Fourier transform infrared spectroscopy (FTIR) confirmed the formation of the metal-oxygen oxide framework. Photocatalytic degradation of MO showed that the unmodified CSO achieved only 39.28% degradation (kapp = 0.0059 min-1), whereas the 1:1 MEA-modified CSO achieved 78.56% degradation (kapp = 0.0124 min-1). Pseudo-first-order kinetics was confirmed for all samples. These results demonstrate that MEA acts as an effective structure-directing and chelating agent that enables precise control over the physicochemical properties of CuSnO3 and substantially enhances its photocatalytic performance.
The effectiveness of various combinations of light-emitting diode (LED) spectra on the in vitro growth of two sugarcane cultivars, KK3 and LK, was investigated. Three steps of micropropagation were investigated: 1) callus induction from young rolled leaves cultured on solid Murashige and Skoog (MS) medium supplemented with 3 mg/L 2,4-dichlorophenoxyacetic acid and 10% coconut water; 2) shoot regeneration from callus cultured on solid MS medium supplemented with 10% coconut water; and 3) the tiller shoots proliferation from in vitro plantlet cultured on MS medium with or without 10% coconut water. Callus induction under blue LEDs followed by shoot regeneration under blue+red LEDs resulted in the highest number of regenerated shoots in KK3 (321.33 shoots/callus clump) followed by callus induction under blue+red LEDs followed by shoot regeneration under white+red LEDs (310.67 shoots/callus clump). In LK, the application of blue+red LEDs during callus induction followed by shoot regeneration under white+red LEDs yielded the highest number of regenerated shoots of 232.33 shoots/callus clump. In contrast, the control treatment-where callus was induced in darkness and shoot regeneration was performed under a white LED alone-produced only 49.00 and 64.33 shoots/callus clump in KK3 and LK, respectively. Furthermore, the white+red LEDs significantly enhanced tiller proliferation in 'KK3'. However, tiller proliferation in 'LK' was not significantly affected by light spectrum.
Plant-based ice cream shows promise as a carrier for probiotic food delivery, supporting both health promotion and environmentally sustainable food systems. Firstly, this study aimed to determine the optimal ratio of water, soy milk and coconut milk using D-optimal mixture design for making plant-based ice cream. Second, a CRD was used to vary the appropriate concentration of Bifidobacterium bifidum Bbi32. The physicochemical characteristics and the sensory properties of the products with and without probiotic supplementation were analyzed. Third, the ice cream after processing was evaluated for cell viability, in vitro gastrointestinal tract, and hydrophobicity. The final product was selected by having probiotics with a not less than 6 log CFU/g and high %hydrophobicity. Lastly, a Logistic Regression analysis was used for consumer acceptance and purchase intention. Results revealed that the optimal formulation was 18% water, 17% soy milk, and 45% coconut milk, which had an overall liking score of 7.40. All probiotic-enriched samples maintained high viable counts (6-8 log CFU/g) after 24 hours of storage at-18 degrees C. Furthermore, all probiotic formulations demonstrated resistance to simulated digestion and exhibited high hydrophobicity (>70%), which may indicate adhesion potential, a key factor in functional food systems. The final product has 100% acceptance rate and 85% purchase intention rate. In conclusion, plant-based ice cream made from soy and coconut milk offers an innovative option for consumers with dairy allergies or those following a vegan diet. Its probiotic properties, combined with the use of sustainable ingredients, contribute to advancing the food industry sector toward improved environmental and health outcomes, aligning with broader goals of food system sustainability.
Rice blast, caused by Magnaporthe oryzae, remains one of the most destructive diseases of rice worldwide. Although molecular diagnostics enable rapid pathogen detection, tools for monitoring avirulence genes that directly inform resistance deployment remain limited. Here, we developed a field-compatible molecular detection platform for the avirulence gene AvrPi9 by integrating rapid crude DNA extraction with recombinase polymerase amplification (RPA) and CRISPR-Cas12a-based detection. The rapid extraction method enabled direct analysis of fungal mycelia and infected rice tissues without conventional DNA purification. Following RPA amplification, AvrPi9 was detected by Cas12a-mediated collateral cleavage and visualized by endpoint fluorescence using a portable blue LED illuminator. The assay specifically detected AvrPi9-positive isolates, with no fluorescence observed in AvrPi9-negative or control samples, and achieved a detection limit of 0.8 ng/& micro;L genomic DNA following RPA amplification. The complete workflow was completed within 40-45 min using only simple heating devices and a portable blue LED illuminator. Validation using experimentally infected and naturally infected field-collected rice leaves confirmed that crude extracts were fully compatible with downstream RPA and CRISPR-Cas12a detection. This platform provides a practical approach for rapid AvrPi9 monitoring to support decentralized disease surveillance, informed resistance gene deployment, and future molecular surveillance of avirulence gene dynamics in M. oryzae populations.
