Abstract
Willaertia magna C2c maky is a thermophilic free-living amoeba strain that showed ability to eliminate Legionella pneumophila, a pathogenic bacterium living in the aquatic environment. The amoeba industry has proposed the use of Willaertia magna as a natural biocide to control L. pneumophila proliferation in cooling towers. Here, transcriptomic and proteomic studies were carried out in order to expand knowledge on W. magna produced in a bioreactor. Illumina RNA-seq generated 217 million raw reads. A total of 8790 transcripts were identified, of which 6179 and 5341 were assigned a function through comparisons with National Center of Biotechnology Information (NCBI) reference sequence and the Clusters of Orthologous Groups of proteins (COG) databases, respectively. To corroborate these transcriptomic data, we analyzed the W. magna proteome using LC–MS/MS. A total of 3561 proteins were identified. The results of transcriptome and proteome analyses were highly congruent. Metabolism study showed that W. magna preferentially consumed carbohydrates and fatty acids to grow. Finally, an in-depth analysis has shown that W. magna produces several enzymes that are probably involved in the metabolism of secondary metabolites. Overall, our multi-omic study of W. magna opens the way to a better understanding of the genetics and biology of this amoeba.
Highlights
Most free-living amoebas (FLAs) are classified in two suprakingdom-level groups: Amoebozoa including Acanthamoeba spp. or Vermamoeba spp. and Excavata comprising Naegleria spp., Vahlkampfia spp. or Willaertia spp. [1,2]
The W. magna trophozoite (50–100 μm in size) form corresponds to the main active stage during which this thermophilic amoeba feeds, moves and divides; it can grow at various temperatures ranging from 22 to 44 ◦C in xenic or axenic culture
Among the genes related to movement, we found that the genes encoding long flagella protein lf4, zinc finger, regulator of chromosome condensation, tubulin and dynein were the most expressed (Table S4)
Summary
Most free-living amoebas (FLAs) are classified in two suprakingdom-level groups: Amoebozoa including Acanthamoeba spp. or Vermamoeba spp. and Excavata comprising Naegleria spp., Vahlkampfia spp. or Willaertia spp. [1,2]. Willaertia magna is a FLA belonging to family Vahlkampfiidae [3]. This protist presents a three-stage life, switching between a trophozoite form, a flagellate form and a cystic form [4,5]. The W. magna trophozoite (50–100 μm in size) form corresponds to the main active stage during which this thermophilic amoeba feeds, moves and divides; it can grow at various temperatures ranging from 22 to 44 ◦C in xenic or axenic culture. The amoeba adopts a cystic form (18–21 μm in size), which allows its survival until the conditions become favorable for cell growth. The transformation to the flagellate stage occurs temporally, being triggered by various stressors [3]
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