| Journal: | Revista do Instituto de Medicina Tropical de Sao Paulo |
| Database: | |
| System number: | 000547857 |
| ISSN: | 0036-4665 |
| Authors: | Martínez Ocaña, Joel1 Martínez Flores, Williams Arony1 Olivo Díaz, Angélica1 Romero Valdovinos, Mirza1 Martínez Hernández, Fernando1 Aguilar Osorio, Guillermo2 Flisser, Ana3 Maravilla, Pablo1 |
| Institutions: | 1Hospital General “Dr. Manuel Gea Gonzalez”, Departamento de Ecología de Agentes Patógenos, Ciudad de México. México 2Universidad Autonoma de México, Instituto de Química, Ciudad de México. México 3Universidad Autonoma de México, Facultad de Medicina, Ciudad de México. México |
| Year: | 2022 |
| Volumen: | 64 |
| Country: | Brasil |
| Language: | Inglés |
| English abstract | Blastocystis sp. is a common intestinal microorganism. The α-L-fucosidase (ALFuc) is an enzyme long associated with the colonization of the gut microbiota. However, this enzyme has not been experimentally identified in Blastocystis cultures. The objective of the present study was to identify ALFuc in supernatants of axenic cultures of Blastocystis subtype (ST)1 ATCC-50177 and ATCC-50610 and to compare predicted ALFuc proteins of alfuc genes in sequenced STs1–3 isolates in human Blastocystis carriers. Excretion/secretion (Es/p) and cell lysate proteins were obtained by processing Blastocystis ATCC cultures and submitting them to SDS–PAGE and immunoblotting. In addition, 18 fecal samples from symptomatic Blastocystis human carriers were analyzed by sequencing of amplification products for subtyping. A complete identification of the alfuc gene and phylogenetic analysis were performed. Immunoblotting showed that the amplified band corresponding to ALFuc (~51 kDa) was recognized only in the ES/p. Furthermore, prediction analysis of ALFuc 3D structures revealed that the domain α-L-fucosidase and the GH29 family's catalytic sites were conserved; interestingly, the galactose-binding domain was recognized only in ST1 and ST2. The phylogenetic inferences of ALFuc showed that STs1–3 were clearly identifiable and grouped into specific clusters. Our results show, for the first time through experimental data that ALFuc is a secretion product of Blastocystis sp., which could have a relevant role during intestinal colonization; however, further studies are required to clarify this condition. Furthermore, the alfuc gene is a promising candidate for a phylogenetic marker, as it shows a conserved classification with the SSU-rDNA gene. |
| Keyword: | Α-L-fucosidase, Blastocystis sp, Glycoside hydrolase, Phylogenetic marker, Subtypes |
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