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Protein and Peptide Letters - Volume 32, Issue 6, 2025

Volume 32, Issue 6

Volume 32, Issue 6, 2025

    • Engineered Bacteriophages: Advances in Phage Genome Redesign Strategies for Therapeutic and Environmental Applications

      https://doi.org/10.2174/0109298665372719250616085616

      Bacteriophages, or phages, have emerged as powerful platforms in synthetic biology, offering innovative solutions for therapeutic and environmental challenges through advanced genome redesign strategies. This review explores a wide range of phage engineering techniques, including CRISPR (clustered regularly-interspaced short palindromic repeats)-Cas systems, phage display, random and site-directed mutagenesis, retrons, and rebooting approaches, highlighting their potential to create phages with tailored functionalities. CRISPR-Cas systems enable precise genome editing, allowing the development of phages with expanded host ranges, biofilm degradation capabilities, and targeted antimicrobial activity. Phage display facilitates the presentation of peptides on phage surfaces, enabling applications in targeted drug delivery, tumor imaging, and bioremediation. Beyond these, techniques like retron-mediated recombination and homologous recombination offer additional avenues for precise phage genome modification. In the therapeutic realm, engineered phages show promise in combating drug-resistant infections, modulating the microbiome, and delivering targeted therapies for cancer and other diseases. Environmentally, phage-based strategies, such as the use of phage-displayed metal-binding peptides, provide innovative solutions for bioremediation and reducing exposure to toxic heavy metals. This review also addresses challenges, such as phage resistance, immune responses, and the limitations of current engineering methods, while exploring future directions, including the development of improved CRISPR systems, phage-based biosensors, and high-throughput screening platforms. By integrating cutting-edge genome redesign strategies with diverse applications, this review underscores the transformative potential of engineered bacteriophages in addressing global healthcare and environmental sustainability challenges.

    • Optimizing Sleep in Athletes: The Potential of α-Lactalbumin in Nutrition Intervention

      Authors: Jingjing Li and Xuepeng Bian
      https://doi.org/10.2174/0109298665363873250623103811

      Athletes frequently encounter sleep deprivation due to the demands of high-intensity training and competition, which can significantly impair their physical recovery and athletic performance. α-Lactalbumin (α-LA), a key component of whey protein that is rich in tryptophan, has been shown to promote the synthesis of serotonin and melatonin, thereby regulating sleep cycles. Moreover, α-LA has demonstrated the ability to reduce inflammation and oxidative stress associated with fatigue and stress, further contributing to improved sleep quality. This review provides a critical evaluation of the current evidence supporting the role of α-LA in enhancing sleep quality in athletes through mechanisms such as neurotransmitter regulation, immune function improvement, and enhancement of antioxidant defenses. Additionally, it highlights the necessity for further research into the differential effects of α-LA on sleep across various sports and gender groups, as well as its potential synergistic interactions with other nutrients. These insights are essential for developing optimized nutritional interventions aimed at enhancing athletic performance.

    • The Interaction between miRNAs and 14-3-3ζ Protein in Different Diseases

      https://doi.org/10.2174/0109298665377739250618153852

      Members of the 14-3-3 protein family are involved in various cellular processes, including migration, angiogenesis, cell cycle, apoptosis, and signal transduction. Nevertheless, the 14-3-3 family possibly plays a fundamental role in the development of diseases and cancer by regulating various biological pathways. MicroRNAs (miRNAs) are mainly transcribed by RNA polymerase II (pol II), with only a few exceptions involving RNA polymerase III (pol III). They can control cell mechanisms through different pathways. miRNAs inhibit or destroy mRNAs by binding to them. They control intracellular mechanisms by binding to molecules such as the 14-3-3ζ protein. miRNAs play a role in regulating this protein, and by inducing or suppressing it, they contribute to either the development or the prevention of the diseases. Therefore, considering the importance of the 14-3-3ζ protein in different pathways within the body, we decided to investigate the relationship between miRNAs and 14-3-3ζ and clarify their interactions, in this review.

    • Effect of Adipokinetic Hormone on Learning-Memory in a Scopolamine-Induced Alzheimer's Model in Mice

      https://doi.org/10.2174/0109298665380324250602054823
      Introduction

      Neurosecretory cells of insects synthesize Adipokinetic Hormone (AKH). Previous studies indicated that AKH improves memory functions. This study aimed to explore the effects of AKH on learning and memory in an Alzheimer's disease model.

      Methods

      Morris Water Maze (MWM), Passive Avoidance (PA), and Modified Elevated Plus Maze (mEPM) tests were conducted in BALB/c mice. Initially, each group consisted of 8 to 9 animals; in total, 120 animals were used in this study. The groups included control, Ani-AKH (1 and 2 mg/kg), Lia-AKH (1 and 2 mg/kg), Pht-HrTH (1 and 2 mg/kg), Scopolamine (1 mg/kg), and Scopolamine combinations. Hormones were given for 6 days in the MWM test to evaluate learning and memory before the second trial in the PA test for memory assessment and after the first trial in the mEPM test to examine consolidation.

      Results

      In the MWM test, Ani-AKH and Pht-HrTH reduced escape latency compared to the scopolamine group (0.05). During the probe trial, Ani-AKH increased time in the escape platform quadrant (0.5) and reversed scopolamine's effects (0.001). Lia-AKH and Pht-HrTh did not affect time in the quadrant but reversed scopolamine's effects (0.01). In the PA test, Ani-AKH reversed scopolamine’s effects (0.5), while Lia-AKH did so in the mEPM test (0.01). The control group showed strong muscarinic receptor staining, while the scopolamine group did not. Ani-AKH and Lia-AKH showed moderate to strong receptor staining, indicating partial restoration.

      Discussion

      AKH and its analogs may enhance memory function by modulating cholinergic pathways, particularly through the partial restoration of muscarinic receptor activity. These results underscore their potential as investigational therapeutics for neurodegenerative disorders characterized by cognitive decline.

      Conclusion

      Our study indicates that AKH may help reduce memory impairments, though the effects depend on the specific assessment methods used in the tests.

    • Recombinant Expression of L-methioninase from and Evaluation of its Anticarcinogenic Properties against MiaPaCa-2 Cells

      https://doi.org/10.2174/0109298665383781250624054915
      Introduction

      This study aimed to investigate the anti-carcinogenic effects of recombinant L- methioninase (rBlmet) on the pancreatic cancer cell line MiaPaCa-2.

      Methods

      In this study, rBlmet was initially cloned, expressed, and purified. To increase enzyme activity, the His-tags on the enzyme were removed using thrombin. rBlmet was then applied to MiaPaCa-2 cells, and the cell viability of MiaPaCa-2 cells was evaluated by neutral red assay after rBlmet treatment. The combined effect of etoposide with rBlmet against MiaPaCa-2 cells was also evaluated for 12 and 24 hours using a neutral red assay. Furthermore, cell morphology was evaluated by Giemsa and DAPI/F-actin staining methods. Survivin and caspase-3 gene expression levels were measured by RT-qPCR.

      Results and Discussion

      The specific activity of the enzyme increased after His-tag elimination to 5.62 µmol/mg per minute. rBlmet showed a significant cytotoxic effect on the MiaPaCa-2 cell line. The IC value (24 h) of rBlmet for MiaPaCa-2 cells was 3.02 U/mL. In addition, rBlmet increased the cytotoxic effect of etoposide on the MiaPaCa-2 cell line, while it showed less effect on HaCat, which is a normal human cell line. Furthermore, rBlmet increased caspase-3 expression and downregulated survivin gene expression in MiaPaCa-2 cell lines. It successfully inhibited the growth of Mia-PaCa-2 cells by exploiting exogenous methionine amino acid in the growth medium. This study revealed promising results. However, further studies are needed on additional pancreatic cancer cell lines and models.

      Conclusion

      Based on these findings, it can be concluded that rBlmet not only has great potential to treat pancreatic cancer in the future but can also be used as an adjuvant to enhance the effectiveness of chemotherapeutic agents like etoposide.

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