|Title||An amoebal grazer of cyanobacteria requires cobalamin produced by heterotrophic bacteria|
|Publication Type||Journal Article|
|Year of Publication||2017|
|Authors||Ma A.T, Beld J., Brahamsha B|
|Journal||Applied and Environmental Microbiology|
|Type of Article||Article|
|Keywords||Amoeba; b-12; corrinoids; dictyostelium-discoideum; gene-expression; identification; lower ligand; microbial interactions; pseudovitamin; social ameba; strain pcc 7002; synthetic biology; vibrio-cholerae; vitamin B-12; vitamin-b-12|
Amoebae are unicellular eukaryotes that consume microbial prey through phagocytosis, playing a role in shaping microbial food webs. Many amoebal species can be cultivated axenically in rich media or monoxenically with a single bacterial prey species. Here, we characterize heterolobosean amoeba LPG3, a recent natural isolate, which is unable to grow on unicellular cyanobacteria, its primary food source, in the absence of a heterotrophic bacterium, a Pseudomonas species coisolate. To investigate the molecular basis of this requirement for heterotrophic bacteria, we performed a screen using the defined nonredundant transposon library of Vibrio cholerae, which implicated genes in corrinoid uptake and biosynthesis. Furthermore, cobalamin synthase deletion mutations in V. cholerae and the Pseudomonas species coisolate do not support the growth of amoeba LPG3 on cyanobacteria. While cyanobacteria are robust producers of a corrinoid variant called pseudocobalamin, this variant does not support the growth of amoeba LPG3. Instead, we show that it requires cobalamin that is produced by the Pseudomonas species coisolate. The diversity of eukaryotes utilizing corrinoids is poorly understood, and this amoebal corrinoid auxotroph serves as a model for examining predator-prey interactions and micronutrient transfer in bacterivores underpinning microbial food webs.