Purification and characterization of cysteine protease of Sarcocystis fusiformis from infected Egyptian water buffaloes
Sarcocystis spp. infects water buffaloes ( Bubalus bubalis ) causing sarcocystosis. In the present study, Sarcocystis fusiformis was recognized in Egyptian water buffaloes based on histological observation and molecular analysis of internal transcribed spacer 1 (ITS1), 18S ribosomal RNA (18S rRNA) a...
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Veröffentlicht in: | Scientific reports 2023-09, Vol.13 (1), p.16123-16123, Article 16123 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | Sarcocystis
spp. infects water buffaloes (
Bubalus bubalis
) causing sarcocystosis. In the present study,
Sarcocystis fusiformis
was recognized in Egyptian water buffaloes based on histological observation and molecular analysis of internal transcribed spacer 1 (ITS1),
18S ribosomal
RNA (18S rRNA) and cytochrome c oxidase subunit I (COX-1) gene fragments. Chemotherapy and vaccines against
Sarcocystis
spp. could potentially target proteases because they may play a crucial role in the infection. Cysteine proteases are multifunctional enzymes involved in vital metabolic processes. However, the involvement of proteases in
S. fusiform
infection has not yet been characterized. Here, the purification and study on some biochemical properties of protease isolated from cysts of
S. fusiform
were carried out. Protease with a molecular weight of 100 kDa was purified. LC–MS/MS analyzed the protein sequence of purified protease and the data suggested that the enzyme might be related to the cysteine protease. The purified protease exhibited maximum activity at pH 6 and a temperature of 50 °C. The Michaelis–Menten constant (K
m
), the maximum velocity (V
max
), and the turnover number (K
cat
) were determined. The complete inhibition effect of cysteine inhibitors indicated that the purified enzyme is a cysteine protease. The results suggested that
S. fusiform
proteolytic enzyme may be necessary for parasite survival in water buffaloes by digesting host tissues. Therefore, cysteine protease could be a suitable target for vaccinations. |
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ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/s41598-023-43147-1 |