Proposed protocol for rate-limiting step determination during anaerobic digestion of complex substrates

Abstract

Anaerobic digestion is a complex process, involving various microorganism groups and, consequently, several reactions. An easy-to-use protocol for the rate-limiting step determination of the process is proposed. The hydrogen production, acetate production, and acetate consumption rates can be calculated, according to a structured algorithm. During the rate limiting step determination, several compounds (biopolymer and monomer representatives, as well as sodium acetate) were used, combined or not with the substrate, to draw the corresponding conclusions. Three substrates were tested, characterized by specific organic compound groups (carbohydrates, proteins, and fats). All three substrates followed the acetate-consuming pathway for the organic matter conversion to methane. In this study, the rate-limiting step for the pathway of acetate consumption was acetate production. Determining the rate-limiting step through the proposed protocol can point to the appropriate actions needed to boost methane production, like substrate pretreatment, using an acidogenic reactor, or checking for the presence of inhibitors.