In this study, various constructs and hosts were used to produce high levels of cecropin B2 (cecB2). To mitigate cecB2's toxic inhibition of host cells, various cecB2 constructs were built. Results showed that the combination of a chitin-binding domain and an intein self-cleavage motif in front of cecropin B2, without a His-tag, was best for cecB2 expression. E. coli ER2566 was the best host, and 2YT was the best medium for cultivation. Under these conditions, a cecB2 yield of 98.2 mg/L could be obtained after purification. The purified cecB2 expressed a wide antimicrobial effect on most Gram-negative strains, including multidrug-resistant Acinetobactor baumannii and Staphylococcus aureus. This study provides a systematic approach to the efficient production of the antimicrobial peptide (AMP) cecB2 via the recombinant E. coli process, which is expected to be an efficient way for the production of other AMPs.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1002/bab.1716 | DOI Listing |
Int J Mol Sci
December 2024
Department of Neurology, Davis School of Medicine, University of California, 1515 Newton Court, Davis, CA 95618, USA.
We have designed and produced 39 amino acid peptide mimics of the and human acetylcholine receptors' (AChRs) main immunogenic regions (MIRs). These conformationally sensitive regions consist of three non-contiguous segments of the AChR α-subunits and are the target of 50-70% of the anti-AChR autoantibodies (Abs) in human myasthenic serum and in the serum of rats with a model of that disease, experimental autoimmune myasthenia gravis (EAMG), induced by immunizing the rats with the electric organ AChR. These MIR segments covalently joined together bind a significant fraction of the monoclonal antibodies (mAbs) raised in rats against electric organ AChR.
View Article and Find Full Text PDFMolecules
December 2024
Department of Chemistry and Life Science, Kogakuin University, Tokyo 192-0015, Japan.
YKL-40 is structurally similar to chitotriosidase (CHIT1), an active chitinase, but it lacks chitin-degrading activity while retaining chitin-binding capability. Elevated YKL-40 levels are associated with inflammatory diseases and cancers, making it a valuable biomarker. We previously reported that the W69T substitution in YKL-40 significantly reduces its chitin-binding affinity, identifying W69 as a crucial binding site.
View Article and Find Full Text PDFACS Appl Mater Interfaces
January 2025
Department of Applied Chemistry, Graduate School of Engineering, Kyushu University, 744 Motooka, Fukuoka 819-0395, Japan.
The rising incidence of fungal infections, compounded by the emergence of severe antifungal resistance, has resulted in an urgent need for innovative antifungal therapies. We developed an antifungal protein-based formulation as a topical antifungal agent by combining an artificial lipidated chitin-binding domain of antifungal chitinase (LysM-lipid) with recently developed ionic liquid-in-oil microemulsion formulations (MEFs). Our findings demonstrated that the lipid moieties attached to LysM and the MEFs effectively disrupted the integrity of the stratum corneum in a mouse skin model, thereby enhancing the skin permeability of the LysM-lipids.
View Article and Find Full Text PDFJ Fungi (Basel)
December 2024
College of Agronomy, Guangxi University, Nanning 530004, China.
Carbohydrate-binding modules (CBMs) are essential virulence factors in phytopathogens, particularly the extensively studied members from the CBM50 gene family, which are known as lysin motif (LysM) effectors and which play crucial roles in plant-pathogen interactions. However, the function of CBM50 in has yet to be fully studied. In this study, we identified seven CBM50 genes from the genome through complete sequence analysis and functional annotation.
View Article and Find Full Text PDFDev Biol
December 2024
University of Rochester, Hutchison Hall, River Campus, Rochester, NY, 14627, USA. Electronic address:
Mollusc shells are diverse in shape and size. They are created by a shell epithelium which secretes a chitinous periostracum membrane at the growing edge of the shell, and then coordinates biomineral deposition on the underside of this membrane. Although mollusc shells are important for studying the evolution of morphology, the molecular basis of the shell development is poorly understood.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!