Abstract

In recent years, the human gut microbiome has been recognised to play a pivotal role in the health of the host. Intestinal homeostasis relies on this intricate and complex relationship between the gut microbiota and the human host. While much effort and attention has been placed on the characterization of the organisms that inhabit the gut microbiome, the complex molecular cross-talk between the microbiota could also exert an effect on gastrointestinal conditions. Blastocystis is a single-cell eukaryotic parasite of emerging interest, as its beneficial or pathogenic role in the microbiota has been a subject of contention even to-date. In this study, we assessed the function of the Blastocystis tryptophanase gene (BhTnaA), which was acquired by horizontal gene transfer and likely to be of bacterial origin within Blastocystis. Bioinformatic analysis and phylogenetic reconstruction revealed distinct divergence of BhTnaA versus known bacterial homologs. Despite sharing high homology with the E. coli tryptophanase gene, we show that Blastocystis does not readily convert tryptophan into indole. Instead, BhTnaA preferentially catalyzes the conversion of indole to tryptophan. We also show a direct link between E. coli and Blastocystis tryptophan metabolism: In the presence of E. coli, Blastocystis ST7 is less able to metabolise indole to tryptophan. This study examines the potential for functional variation in horizontally-acquired genes relative to their canonical counterparts, and identifies Blastocystis as a possible producer of tryptophan within the gut.

Highlights

  • Blastocystis is a highly prevalent stramenopilic parasite of the gastrointestinal system, estimated to be present in over one billion humans worldwide [1], along with a wide variety of mammalian, reptilian, amphibian, and avian hosts [2]

  • We show that canonical E. coli K12 TnaA has limited homology with BhTnaA, while the gene is highly conserved between Blastocystis ST 1, 4, and 7

  • Human Blastocystis isolate ST7-B was acquired from a patient at the Singapore General Hospital in the early 1990s, before the Institutional Review Board was established at the National University of Singapore (NUS)

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Summary

Introduction

Blastocystis is a highly prevalent stramenopilic parasite of the gastrointestinal system, estimated to be present in over one billion humans worldwide [1], along with a wide variety of mammalian, reptilian, amphibian, and avian hosts [2]. Blastocystis strains are classified into subtypes (ST) of the Blastocystis genus [3], rather than species. Genomic and ribosomal sequencing have identified 17 Blastocystis subtypes, designated ST1 through ST17 [4]. ST1 through 9 are found in humans [5]. Complete genomic sequencing data is available for ST1, 4, and 7. ST7, used in this paper, is classified into various isolates, denoting their clinical origin [7]. This study focuses on subtype 7, isolate B, referred to as ST7-B

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