title: Evidence of isoprenoid precursor toxicity in Bacillus subtilis.
abstract: The mevalonic acid (MVA) and methylerythritol phosphate (MEP) pathways for isoprenoid biosynthesis both culminate in the production of the two-five carbon prenyl diphosphates: dimethylallyl diphosphate (DMAPP) and isopentenyl diphosphate (IPP). These are the building blocks for higher isoprenoids, including many that have industrial and pharmaceutical applications. With growing interest in producing commercial isoprenoids through microbial engineering, reports have appeared of toxicity associated with the accumulation of prenyl diphosphates in Escherichia coli expressing a heterologous MVA pathway. Here we explored whether similar prenyl diphosphate toxicity, related to MEP pathway flux, could also be observed in the bacterium Bacillus subtilis. After genetic and metabolic manipulations of the endogenous MEP pathway in B. subtilis, measurements of cell growth, MEP pathway flux, and DMAPP contents suggested cytotoxicity related to prenyl diphosphate accumulation. These results have implications as to understanding the factors impacting isoprenoid biosynthesis in microbial systems.
authors.affiliation: Department of Chemistry, Saginaw Valley State University, University Center, MI 48710, USA. tsivy@svsu.edu
authors.forename: Tami L
authors.initials: TL
authors.lastname: Sivy
authors.forename: Ray
authors.initials: R
authors.lastname: Fall
authors.forename: Todd N
authors.initials: TN
authors.lastname: Rosenstiel
mesh_terms: D001412:Bacillus subtilis / Q000166:cytology / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D002070:Butadienes; D019748:Carbon-Carbon Double Bond Isomerases / Q000172:deficiency / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D003603:Cytotoxins / Q000096:biosynthesis*; D004896:Erythritol / Q000031:analogs & derivatives / Q000378:metabolism; D005578:Fosfomycin / Q000031:analogs & derivatives / Q000494:pharmacology; D005818:Genetic Engineering; D045782:Hemiterpenes / Q000096:biosynthesis; D009943:Organophosphorus Compounds; D010420:Pentanes; D017384:Sequence Deletion; D013403:Sugar Phosphates / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism*
chemical_list: C114232:2-C-methylerythritol 4-phosphate; D002070:Butadienes; D003603:Cytotoxins; D045782:Hemiterpenes; D009943:Organophosphorus Compounds; D010420:Pentanes; D013403:Sugar Phosphates; D013729:Terpenes; C005059:isoprene; D005578:Fosfomycin; C004809:isopentenyl pyrophosphate; C043060:3,3-dimethylallyl pyrophosphate; C024640:fosmidomycin; D019748:Carbon-Carbon Double Bond Isomerases; C021453:isopentenyldiphosphate delta-isomerase; D004896:Erythritol