Development of therapies that selectively inhibit cancer cells but leave normal cells untouched is an important impediment to progress in cancer therapy. This paper reports the novel strategy employing the R-Tf-D-LP4 peptide, an unconventional cell penetrating peptide selectively targeting liver cancer cells, combined with the potent antitumor cytokine interleukin 15 (IL-15), with selective activity towards liver cancer cells. The fusion protein was constructed as a chimera between the IL-15 and the targeting peptide R-Tf-D-LP4 to improve the targeting specificity of this protein to the desired cells. With the help of modern computer modeling technologies, a fusion protein was designed, and its interactions with the IL-15 receptor complex (IL-15Rα/IL-15Rβ/γc) were evaluated using a molecular docking technique. The structure and physicochemical properties of the chimeric protein were determined using programs such as trROSETTA, I-TASSER, and ProtParam, after which they were further verified using SAVES 6.1 server with an excellent ERRAT quality score of 99.35
Squalene, a naturally occurring triterpenoid, is widely utilized in the food, pharmaceutical, and vaccine adjuvant industries. Conventional production methods, which rely on extraction from shark liver oil or plant sources, are limited by ecological sustainability concerns and low extraction efficiency. Recent progress in synthetic biology and metabolic engineering has enabled the development of sustainable and efficient microbial platforms for squalene biosynthesis. This review provides a comprehensive overview of the biosynthetic pathways of squalene in yeast, summarizes key metabolic engineering strategies, and highlights recent advancements in the field, with an emphasis on multi-organelle synergistic engineering and critical distinction of evidence sources. We focus on approaches such as enhancing the mevalonate pathway, modulating competing metabolic routes, applying subcellular compartmentalization engineering, optimizing cofactor balance, and implementing product efflux strategies. Notably, integrated strategies, particularly organelle engineering in Yarrowia lipolytica and Saccharomyces cerevisiae, have increased squalene titers to high levels (up to 55.8 g/L). These advances provide a useful basis for future metabolic engineering efforts aimed at improving squalene production and facilitating the synthesis of related terpenoids.
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.
