This review examines Geobacillus sensu stricto as a source of thermostable glycoside hydrolases (GH) for biomass conversion, food processing, and enzyme engineering. Recent peer-reviewed literature was assessed with emphasis on taxonomy, genome-based Carbohydrate-Active Enzymes (CAZyme) prediction, biochemical validation, structural data, and engineering case studies. Taxonomic boundaries were interpreted using current Anoxybacillaceae frameworks, with Parageobacillus treated as a related comparator rather than as Geobacillus. The strongest evidence supports GH13 alpha-amylases, xylan-active systems, beta-xylosidases, and selected accessory enzymes. Recent studies also show that genome mining must be coupled with enzymatic assays and product profiling because CAZyme annotation alone does not prove industrial function. Molecular engineering has improved relevant traits, including the longer thermal half-life of engineered G. stearothermophilus alpha-amylase variants, the increased catalytic efficiency of oligo-alpha-1,6-glucosidase variants, and improved AmyS expression in Bacillus subtilis. Geobacillus glycoside hydrolases are best interpreted as process-specific, engineerable biocatalytic templates. Their translation requires reliable taxonomy, functional validation, structural interpretation, scalable expression and testing on realistic substrates. This synthesis also recognises current limitations: many predicted CAZymes still lack biochemical validation, complete cellulolytic systems remain less mature than xylan- and starch-active systems, and scale-up data remain scarce.
Epigenetic regulation refers to heritable changes in gene expression that occur without alteration of the DNA sequence, primarily mediated by DNA methylation. In this study, we applied two DNA methyltransferase (DNMT) inhibitors, zebularine (Zeb) and 5-azacytidine (5-Azac), to alter the epigenetic landscape of Dunaliella salina and observe the resulting phenotypic changes. Long-term (40-day) treatment revealed that, on Day 40, Zeb dose-dependently increased the proportion of 9-cis β-carotene from 52.4
Mycotoxins, especially aflatoxins produced by fungi of the genus Aspergillus, threaten global food security and public health due to their carcinogenic and immunosuppressive effects. To exploit Key enzymatic steps in the aflatoxin biosynthetic pathway for therapeutic applications, the aflQ gene (also Known ordA) -wich encodes acytochrome P450 monooxygenase capable of generating reactive oxidative species -was targeted as a candidate antimicrobial agent. This study aims to extract and characterize the aflQ gene (known as ordA) from the Aspergillus niger fungus isolated from grain silos in Anbar Governorate, ands proteomically evaluate it as an innovative antimicrobial. The methodology involved isolating the fungus and molecularly characterizing it by amplifying and sequencing a segment ofthe18S rRNA gene. The species identity was confirmed utilizing the BLAST tool in the NCBI database. The sequence was deposited in GenBank and received an Accession number (PV455323). Subsequently, amplification of the aflQ gene was performed using polymerase chain reaction (PCR) and cloned into the pET-30b( +) expression vector. The hybrid plasmid was transformed into E. coli BL21 bacteria, and gene expression was induced using IPTG. The protein production was confirmed by SDS-PAGE analysis at the predicted molecular weight of 63–64 kDa. The results showed that the recombinant AflQ protein exhibited significantly enhanced inhibitory potency, with inhibition diameters reaching 24 mm against Gram-positive bacteria (Staphylococcus and Enterococcus) and 12.5 mm against Gram-negative bacteria (E. coli and K. pneumoniae). Conversely, the control bacterial culture showed no inhibitory activity, verifying that the observed efficacy specific to the AflQ protein. The study concludes that the AflQ protein represents a new class of oxidoreductase/Cytochrome P450antimicrobial agents. It highlights the significance of utilizing cellular debris to reduce industrial production costs by eliminating expensive purification steps.
Staphylococcus aureus is a major human pathogen causing diseases ranging from mild skin infections to life-threatening conditions due to its diverse virulence factors. The emergence of antibiotic-resistant strains underscores the urgent need for an effective vaccine, though none has yet achieved clinical success. The nasal mucosa serves as a natural reservoir in about 30
Drug-induced cardiotoxicity remains a significant challenge in pharmaceutical development. While patch-clamp remains the gold standard for evaluating ion channel effects, its low throughput limits early-phase screening. Multi-electrode array (MEA) systems offer higher throughput but often fail to mechanistically distinguish between various sodium channel modulators, such as agonists versus antagonists. The purpose of this study was to develop a cardiac safety assessment method capable of mechanistically distinguishing sodium channel modulators. In this study, we developed a cardiac safety assessment method using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) combined with fluorescence-based intracellular Na+ flux imaging. We characterized hiPSC-CM batches by verifying the expression of key ion channel genes, including SCN5A, ATP1A1, and SLC8A1. Functional responses to sodium channel modulators were evaluated using MEA analysis. Intracellular Na+ intensity changes were quantified using the sodium sensitive dye CoroNa Green. MEA analysis confirmed functional responses to various Nav1.5 inhibitors and activators, though distinguishing specific modes of action was limited. By utilizing the sodium-sensitive dye CoroNa Green, we successfully quantified real-time Na+ intensity changes. Our results demonstrate that Nav1.5 activators (e.g., Aconitine) and Na+/K+-ATPase inhibitors (e.g., Ouabain, Digoxin) significantly increase intracellular Na+ levels at both 1-h and 24-h intervals. This imaging-based approach provides a mechanistic tool for distinguishing sodium channel modulators, offering an alternative for next-generation cardiovascular safety pharmacology.
