Estimating changes of isotopic fractionation based on chemical kinetics and microbial dynamics during anaerobic methane oxidation: apparent zero- and first-order kinetics at high and low initial methane concentrations.

Changes in natural isotopic composition may be used to reveal metabolic pathways of substrate transformation by microbial communities (Vavilin in Ecol Model 240:84-92, 2012b). Anaerobic oxidation of methane (AOM) by sulfate has been described using a mathematical model based on chemical kinetics, microbial dynamics and equations for (13)C isotope accumulation in products as well as their redistribution between substrate and products. Experimental data for two batch cultures that originated from microbial mats covering methane seep chimneys in the Black Sea, previously obtained by Seifert et al. (Org Geochem 37:1411-1419, 2006) and Holler et al. (Env Microbiol Reports 1(5):370-376, 2009), were used to model AOM. During long-time incubation, changes of isotope signatures in CH(4) showed that in the Seifert et al. batch tests (low methane concentration), in contrast to the Holler et al. batch tests (high methane concentration), methane production occurred along with methane oxidation. In accordance with the model, apparent zero and first-order kinetics of methane oxidation were valid for the Holler et al. and Seifert et al. batch tests, respectively. The observed change of [Formula: see text] was explained by microbial kinetics reflecting that the rate is lower for heavy substrate microbial utilization when compared to light substrate microbial utilization. The model showed that small amounts of methanogenesis will change the carbon isotopic composition of methane because biogenic methane has a distinct isotopic composition and due to the large difference between the maximum specific rates of methane oxidation and production. The estimated biomass doubling time of methane-oxidizers for high and low methane concentration was 408/126 days and 4640/1160 days, respectively, depending on the value of the half-saturation constant K ( S ) (5 and 20 mM).