چکیده مقاله
The escalating demand for sustainable protein sources has propelled the exploration of plant based meat alternatives, where functional enhancements are pivotal for mimicking animal derived textures and nutritional profiles Central to this innovation is the precise manipulation of amino acid isomers, particularly glycine variants, which influence protein folding, gelation, and sensory attributes in meat analogs This study introduces a novel hybrid polymeric nanomembrane engineered through genetic modification and electron beam irradiation, integrated with thermophilic Bacillus derived enzymes for the stereospecific separation and concentration of glycine isomers The nanomembrane, fabricated from a chitosan polyvinyl alcohol PVA matrix reinforced with silica nanoparticles, leverages electron beam crosslinking to achieve nanoscale porosity 10 50 nm and enhanced mechanical stability, ensuring biocompatibility and selectivity for biomolecular filtration Genetic engineering targeted the glycine oxidase GO gene from Bacillus thermophilus, a thermophilic strain resilient to high temperature processing, to produce a mutant enzyme with heightened specificity for D glycine over L glycine Site directed mutagenesis at key residues e g , Ala255Ser amplified enantioselectivity by 3 5 fold, as validated through in vitro assays The recombinant enzyme was immobilized onto the nanomembrane surface via covalent linkage to glutaraldehyde activated chitosan, facilitating continuous flow separation Experimental protocols encompassed membrane synthesis, enzyme expression in Escherichia coli BL21 DE3 , and chromatographic validation using high performance liquid chromatography HPLC for isomer purity assessment Performance metrics revealed exceptional separation efficiency: under optimized conditions pH 8 0, 50°C, flow rate 1 mL/min , the system achieved 92% enantiomeric excess ee for D glycine concentration from a racemic mixture, with flux rates of 45 L/m² h and rejection coefficients exceeding 98% for non target solutes Concentrated D glycine fractions, enriched to 85% purity, were incorporated into soy based meat analogs, enhancing textural firmness by 28% measured via texture profile analysis and umami perception via Maillard reaction precursors Nutritional profiling indicated a 15% uplift in essential amino acid bioavailability, corroborated by simulated gastrointestinal digestion models This integrated platform addresses critical bottlenecks in food biotechnology: scalability, enzyme stability under industrial shear, and eco friendly membrane disposal By harnessing thermophilic enzymes' robustness optimal activity at 60°C, half life >120 min , the system outperforms mesophilic counterparts, reducing energy inputs by 40% in downstream processing Furthermore, the nanomembrane's antifouling properties, derived from hydrophilic PVA domains, minimized biofouling by 65%, extending operational lifespan to 500 cycles Challenges such as membrane flux decline post 100 cycles attributed to enzyme desorption were mitigated through periodic regeneration with 0 1 M NaCl washes, restoring 88% initial performance Economic feasibility analysis projected a 25% cost reduction in glycine isomer production compared to synthetic routes, positioning this technology as a cornerstone for functional food fortification Broader implications extend to precision nutrition, where isomer specific delivery could modulate gut microbiome interactions, fostering anti inflammatory profiles in plant based diets In summary, this work pioneers a synergistic fusion of genetic engineering, nanomaterials, and bioprocessing to unlock glycine isomers' potential in plant based meats, bridging sustainability with sensory innovation Future iterations may incorporate CRISPR edited Bacillus strains for on site enzyme production, amplifying industrial viability This advancement not only elevates plant based protein efficacy but also exemplifies bio inspired solutions for global food security amid climate constraints
کلیدواژهها
نویسندگان
Reyhane Ahmadi
Ph D Student in Food Science and Technology, Department of Chemical Technologies, Iranian Research Organization for Science and Technology IROST , Tehran, Iran
Morteza Jamshid Eini
PhD in Food Technology, Islamic Azad University, North Tehran Branch, Tehran, Iran
Behzad Beizaei
PhD student in Food Technology, Islamic Azad University, Tehran Azad Medical Sciences Branch, Tehran, Iran
Parisa Moallemi
PhD student in Food Science and Technology Engineering, Islamic Azad University, Noor Branch, Mazandaran, Iran
شیوه ارجاع
Ahmadi, Reyhane and Jamshid Eini, Morteza and Beizaei, Behzad and Moallemi, Parisa,1404,Genetic Engineering of a Hybrid Polymeric Nanomembrane via Electron Beam Irradiation,The first national conference environment, smart agriculture, food security,Varamin
ارائهشده در
مجموعه مقالات اولین همایش ملی محیط زیست، کشاورزی هوشمند، امنیت غذایی28 آبان 1404 · ورامین