Intracellular carbon isotope distributions of continuous-culture Allochromatium vinosum grown on acetate vs. CO2.

Sulfur oxidizing bacteria are commonly observed in various aquatic environments, which use reduced sulfur compounds as electron donors to complete the carbon metabolism. Their carbon isotope fractionations during biosynthesis can be preserved in the fossil records. Here we grew a model sulfur oxidizing bacterium, Allochromatium vinosum, in two mode of continuous culture. One of the A. vinosum culture was grown autotrophically with CO₂ as the carbon source; while the other one was grown heterotrophically on acetate. A novel protein isotope fingerprinting analysis was applied combining proteomics and protein isotope analysis together, which indicates no isotope fractionation among individual proteins, whereas the bulk protein d¹³C relative to bulk biomass were substantially different between autotrophic and heterotrophic cells. The same trend was also observed in d¹³C values of bulk amino acids, fatty acids and nucleic acids. The observed difference in major classes of organic compounds may result from the difference in biosynthetic pathways of autotrophic and heterotrophic cells. A closer look into d¹³C value of individual amino acids and fatty acids provides us further evidence to identify isotopic response to key reactions of central carbon metabolism as revealed by proteomic analysis. Our work suggests that we can decipher diverse microbial carbon metabolisms by combining proteomics with compound specific analysis of major classes of organic compounds.