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- Title
Recombinant TP-84 Bacteriophage Glycosylase–Depolymerase Confers Activity against Thermostable Geobacillus stearothermophilus via Capsule Degradation.
- Authors
Łubkowska, Beata; Sobolewski, Ireneusz; Adamowicz, Katarzyna; Zylicz-Stachula, Agnieszka; Skowron, Piotr M.
- Abstract
The TP-84 bacteriophage, which infects Geobacillus stearothermophilus strain 10 (G. stearothermophilus), has a genome size of 47.7 kilobase pairs (kbps) and contains 81 predicted protein-coding ORFs. One of these, TP84_26 encodes a putative tail fiber protein possessing capsule depolymerase activity. In this study, we cloned the TP84_26 gene into a high-expression Escherichia coli (E. coli) system, modified its N-terminus with His-tag, expressed both the wild type gene and His-tagged variant, purified the recombinant depolymerase variants, and further evaluated their properties. We developed a direct enzymatic assay for the depolymerase activity toward G. stearothermophilus capsules. The recombinant TP84_26 protein variants effectively degraded the existing bacterial capsules and inhibited the formation of new ones. Our results provide insights into the novel TP84_26 depolymerase with specific activity against thermostable G. stearothermophilus and its role in the TP-84 life cycle. The identification and characterization of novel depolymerases, such as TP84_26, hold promise for innovative strategies to combat bacterial infections and improve various industrial processes.
- Subjects
GEOBACILLUS stearothermophilus; ESCHERICHIA coli; LIFE cycles (Biology); GENOME size; GENETIC variation; BACTERIOPHAGES
- Publication
International Journal of Molecular Sciences, 2024, Vol 25, Issue 2, p722
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms25020722