BiotechIntellect

Nutritional Reformulation to Functional Food Innovation: An Evidence-Informed Framework for the Development of Synbiotic Cereal-based Snacks

Mohammad Mahdi Fadavi iD , Samira Hoseini iD , Ghasem Fadavi iD , Fatemeh Mohammadi-Nasrabadi iD , Fatemeh Esfarjani ^* iD

https://doi.org/10.66224/BiotechIntellect.3.1.62

Background: Commercial snack foods are widely consumed but may provide excessive saturated and trans fatty acids, refined carbohydrates, and energy while contributing limited dietary fiber. This study aimed to develop a health-oriented symbiotic cereal-based snack integrating nutritional reformulation with probiotic-prebiotic functionality.

Material and Methods:An evidence-informed conceptual framework was developed by synthesizing scientific and regulatory evidence across seven domains: dietary fat quality, carbohydrate quality, probiotics, prebiotics, synbiotics, food-matrix and processing considerations, and health-claim substantiation. Evidence was translated into sequential product-development and validation stages addressing lipid quality, whole grains and dietary fiber, strain-specific probiotic selection, prebiotic functionality, synbiotic integration, and probiotic protection and stability.

Results: The framework establishes a progressive evidence hierarchy: nutritional reformulation → functional-ingredient characterization → product stability and safety → demonstrated synbiotic functionality → substantiated health benefit. Required validation includes strain-level identification and safety, probiotic viability throughout processing and storage, prebiotic selective utilization, physicochemical stability, sensory acceptance, microbiological safety, and appropriate human evidence when health benefits are claimed. The framework emphasizes that prebiotic co-incorporation alone is insufficient to demonstrate synbiotic functionality.

Conclusion: A symbiotic cereal-based snack provides a practical researchdevelopment model for integrating nutritional improvement with functional-food innovation. The proposed framework offers a translational pathway for evidence-based functional snack development while supporting scientifically appropriate product characterization, regulatory evaluation, and substantiation of synbiotic and health claims.

Screening of Pigment Producer from Native Halophilic Archaea

Milad Behroozi , Maryam Jalili Tabaii iD , Soheila Abbasi iD , Giti Emtiazi iD , Ramona Massoud iD

https://doi.org/10.66224/BiotechIntellect.3.1.58

Purpose and aim: Halophilic archaea have a high potential to discover new compounds with high biotechnological potential, one of these compounds is carotenoids.

Material and Methods: A collection of 23 native highly halophilic archaea isolates were isolated from distinct saline habitats in different regions of Iran. Among the examined isolates, the isolates that had produced colored colonies within a week were further investigated by absorption spectrophotometric method in order to extract and measure the amount of pigment produced with different solvents of methanol, acetone, hexane, ethernet and ethanol. UV, TLC, FT-IR, LC-MS spectroscopic methods were used to investigate and determine the carotenoid pigment structure of Haloferax Mediterranei ASFM403 strain. Its antioxidant and antimicrobial properties were investigated. Also, the effect of produced carotenoid toxicity on melanoma cancer cells and human fibroblast cells was studied

Results and Discussion: According to the results obtained from the screening of the strains, the best methanol solvent and the best absorbance 0.239 at the wavelength of 498 belonged to the Haloferax Mediterranei ASFM403 strain. The light absorption data of the pigment shows the structural similarity of this carotenoid with bacterioruberin.

Conclusion: The information obtained from carotenoid identification with TLC, FT-IR and LC-MS shows that bacterioruberin is the dominant carotenoid. The antioxidant activity of carotenoid obtained at a concentration of 5 μg reached 83%, which was higher than ascorbic acid as a standard. And the antibacterial effect was significant against pathogenic bacteria. It also has a toxic effect on melanoma cancer cells in concentrations of 10 and 50 micrograms/liter. While, it helps the growth of fibroblast cells.

Multi-Targeting the DNA Damage Response in Breast Cancer: An In Silico Study of Zerumbone–Protein Interactions

Zahra Sanaimehr iD , Nasim Forgani iD , Nima Ghahremani Nezhad ^* , Ahmed Issa Al-Tameemi iD

https://doi.org/10.66224/BiotechIntellect.3.1.60

Background and Objective: Breast cancer remains a major cause of cancer-related mortality among women, with tumor heterogeneity and therapeutic resistance presenting significant challenges to treatment. The DNA damage response (DDR) pathway is essential for DNA repair and may contribute to cancer cell survival and treatment resistance, making DDR-associated proteins potential therapeutic targets. This study investigated the potential interactions of zerumbone, a bioactive compound derived from Zingiber zerumbet, with DDR-related proteins using molecular docking, molecular dynamics (MD) simulations, and molecular mechanics Poisson–Boltzmann surface area (MM/PBSA) analysis.

Materials and Methods: Molecular docking of 12 DDR-related protein structures identified favorable predicted interactions, with docking scores ranging from −5.24 to −6.69 kcal/mol. Five complexes with the most favorable docking scores were selected for three independent 100-ns MD simulations.

Results and Conclusion: Root-mean-square deviation and fluctuation (RMSD & RMSF) analyses revealed differences in global structural deviations and residue-level flexibility, with 3JVE–zerumbone exhibiting relatively low RMSD values and 7CMZ–zerumbone showing greater structural fluctuations. Radius of gyration analyses indicated that overall structural compactness remained relatively consistent during the simulations, while principal component analysis revealed comparatively restricted conformational sampling for 3JVE–zerumbone and broader conformational exploration for 7CMZ–zerumbone. MM/PBSA calculations, performed without an entropy term, yielded estimated binding energies ranging from −8.20 to −26.53 kcal/mol, with 7CMZ–zerumbone and 3AL2–zerumbone exhibiting the most favorable estimates. Notably, 3JVE–zerumbone displayed relatively low RMSD values but the least favorable MM/PBSA estimate, illustrating that limited structural deviation does not necessarily correspond to more favorable estimated binding energetics. Collectively, these findings provide structural and energetic insights into the predicted interactions of zerumbone with DDR-related protein domains and identify candidate complexes for further investigation.

Biotechnological Mitigation of Phthalates in the Food Chain: Packaging Migration, Dietary Exposure, and Probiotic Detoxification

Mohammad Shakeri ^* , Fatemeh Shakeri iD , Runak Ghobadi iD , Fatemeh Gholami iD

https://doi.org/10.66224/BiotechIntellect.3.1.55

Background and aims: Phthalates are plasticizers produced in volumes so large that dietary exposure has become universal. Because they remain unbound within polymer networks, these compounds migrate into food during processing, storage, and distribution, with additional inputs entering crops and livestock through environmental contamination.

Materials and Methods: Following a structured search across four databases, this review synthesizes evidence on the routes of food contamination, analytical challenges, dietary intakes, toxicological endpoints, and emerging mitigation strategies.

Results and Conclusion: Occurrence data consistently show that lipid-rich items such as edible oils and dairy products accumulate the heaviest loads, though storage temperature and acidity can also drive measurable leaching into bottled water and soft drinks. Estimated intakes indicate that young children and high consumers of packaged foods absorb a noticeable share of established tolerable daily intakes, particularly when compounds are assessed as cumulative mixtures under European Food Safety Authority guidance. Given that current regulations and industrial substitutes like DINCH and DEHT leave historical residues in circulation, biological mitigation has emerged as a complementary safeguard. Food-grade lactic acid bacteria, notably strains of Lactobacillus and Lactiplantibacillus, can physically sequester phthalate diesters and reactive monoesters such as MEHP through peptidoglycan and exopolysaccharide surface binding, reducing free toxicant fractions in fermented matrices and accelerating fecal clearance in vivo. We conclude by outlining critical research priorities, including outdated occurrence surveys, single-compound regulatory baselines, and the need to validate microbial biosorption under commercial food-processing conditions.

Production and Partial Purification of Pectinase from a Mutant of Penicillium EZ-ZH390 in Solid State Fermentation

Zohreh Hamidi-Esfahani iD , Rezvan Mostofi

https://doi.org/10.66224/BiotechIntellect.3.1.56

Background and Aims: the biotechnological potential of pectinolytic enzymes from fungi has drawn a great deal of attention from various researchers worldwide. Various agro-industrial wastes such as wheat bran, sugarcane bagasse, coffee pulp, lemon peel and apple pomace have been explored for the microbial production of pectinase

Material and Methods: This study describes pectinase production using Penicillium sp. EZ-ZH390 in solid-state fermentation by Plackett–Burman design (PBD) and one-factor-at-a-time (OFAT) methodology. Four variables (moisture content, inoculum size, particle size, MgSO₄ 7H₂O), which showed the most effective factors by the PBD analysis. Moisture and MgSO₄ 7H₂O had the most positive impact on pectinase production further optimized using the OFAT method.

Results and Discussion: The optimization results indicated that a maximal pectinase activity of 3825 U/gds was achieved at, 85% initial moisture content and using 0.1% (w/v) MgSO₄ 7H₂O, particle size smaller than 0.7 mm and 10⁸ spores/mL in the fermentation medium. The results showed a 1.92-fold increase in pectinase activity of Penicillium sp. EZ-ZH390 under the optimized conditions. Orange peel can be considered as low-cost substrate for pectinase production. Ethyl alcohol and acetone at different concentrations were then used for ethanol-precipitated enzyme. Results showed that pectinase was partially purified 14.37-fold using 60% ethyl alcohol. The optimum pH and temperature of the crude and partially purified enzyme were 5, 55 °C and 5, 45 °C respectively.

Activity of a Carbohydrate-rich Extracellular Fraction from Ganoderma lucidum

Fariba Soleimani , Ashrafsadat Hatamian-Zarmi , Fatemeh Yazdian ^* , Nima Ghahremani Nezhad iD

https://doi.org/10.66224/BiotechIntellect.3.1.52

Background and aims: In this study, the antimicrobial activity of a carbohydrate-rich extracellular fraction produced from submerged cultures of Ganoderma lucidum was investigated together with a statistical evaluation of culture conditions.

Materials and Methods: Five culture-related factors, including malt extract, yeast extract, pH, methyl jasmonate (MeJA), and the phosphate-source system, were screened using a Plackett–Burman design. pH and MeJA concentration were subsequently evaluated by response surface methodology using MIC as the biological response.

Results and Conclusion:The model-derived operating condition was pH 6.36 and 14.24 µg/L MeJA. Under the investigated conditions, the recovered extracellular fraction showed antibacterial activity against Bacillus subtilis, Bacillus cereus, Salmonella typhi, and E. coli. The lowest MIC was observed for B. subtilis (30 mg/mL), whereas E. coli showed lower susceptibility, with a MIC of 75 mg/mL under the selected condition. The total carbohydrate content of the recovered dry fraction was 825.30 µg glucose equivalents per mg. These findings show that culture conditions, particularly pH and MeJA concentration, influence the antibacterial performance of the recovered carbohydrate-rich extracellular fraction.

An Amperometric Biosensor Based on Cytochrome C Immobilized on a Gold Electrode for Detection of Dissolved Hydrogen Sulfide

Soraya Bahari , Fatemeh Yazdian ^* iD , Ashrafsadat Hatamian-Zarmi iD , Majid Kalatebojdi , Nima Ghahremani Nezhad iD

https://doi.org/10.66224/BiotechIntellect.3.1.53

Background and aims: Cytochrome C, a metalloprotein, can play the role of electron transporter electrochemically. In the present work, an amperometric biosensor for the detection of hydrogen sulfide in aqueous media has been fabricated by immobilizing Cytochrome C on the surface of gold electrode.

Materials and Methods: The surface was modified by nanoparticle and self-assemble monolayer of 3- mercaptopropionic acid. 1-(3-DimethylAminoPropyl)-3-EthylCarboDiimide Methiodide (EDC) and N-hydroxysuccinimide (NHS) were applied as coupling reagents for covalent attachment of Cytochrome C on modified gold electrode surface.

Results and Conclusion: Experimental design based on Central Composite Design (CCD) method revealed the optimal condition for current peak at pH 6, 5 mM phosphate buffer and 20 °C. In the presence of hydrogen sulfide in electrolyte, oxidation and reduction peaks reduced and over time were completely vanished. Under the investigated aqueous conditions, the lowest tested H₂S concentration was 0.001 ppm, and a rapid amperometric response was observed. The electrode response remained observable during the investigated 15-day period.

Synbiotic Yoghurt Enriched with Lactobacillus acidophilus, Long-Chain Inulin, and Moringa oleifera Leaves Extract

Soroush Orouji ^* , Zahra Beigmohammadi iD , Hamid B. Ghoddusi

https://doi.org/10.66224/BiotechIntellect.3.1.54

Background and aims: In today's world, the development of functional foods has become of paramount importance. This research aimed to produce a synbiotic yogurt enriched with Lactobacillus acidophilus, long-chain inulin, and Moringa oleifera extract.

Materials and Methods: Two levels of Moringa oleifera Leaves Extract (1 and 2% w/w) and inulin (3 and 5% w/w) were tested for their effects on the properties of the synbiotic yogurt over 20 days of storage at 4°C.

Results and Conclusion: The level of the extract had a profound impact on both acidity and pH, increasing the former and decreasing the latter. The extract increased yogurt's antioxidant activity compared to the control. Inulin had the most notable effect on viscosity, syneresis, water-holding capacity, and dry matter content. The yogurt sample containing 1% extract and 5% inulin received the highest sensory scores. Inulin improved the viability of probiotic bacteria, while Moringa oleifera extract did not affect any aspect of them. Neither Inulin nor Moringa oleifera extract significantly affected the fat content. This new synbiotic product, incorporating Lactobacillus acidophilus, M. oleifera extract, and inulin, possesses the desired characteristics and could effectively deliver probiotic strains and bioactive compounds together, thereby providing potential health benefits.

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