Abstract
Medium-chain carboxylates such as n-caproate and n-caprylate are valuable chemicals, which can be produced from renewable feedstock by anaerobic fermentation and lactate-based microbial chain elongation. Acidogenic microbiota involved in lactate-based chain elongation and their interplay with lactic acid bacteria have not been characterized in detail yet. Here, the metabolic and community dynamics were studied in a continuous bioreactor with xylan and lactate as sole carbon sources. Four succession stages were observed during 148 days of operation. After an adaptation period of 36 days, a relatively stable period of 28 days (stage I) was reached with n-butyrate, n-caproate and n-caprylate productivities of 7.2, 8.2 and 1.8 gCOD L–1 d–1, respectively. After a transition period, the process changed to another period (stage II), during which 46% more n-butyrate, 51% less n-caproate and 67% less n-caprylate were produced. Co-occurrence networks of species based on 16S rRNA amplicon sequences and correlations with process parameters were analyzed to infer ecological interactions and potential metabolic functions. Diverse functions including hydrolysis of xylan, primary fermentation of xylose to acids (e.g., to acetate by Syntrophococcus, to n-butyrate by Lachnospiraceae, and to lactate by Lactobacillus) and chain-elongation with lactate (by Ruminiclostridium 5 and Pseudoramibacter) were inferred from the metabolic network. In stage I, the sub-network characterized by strongest positive correlations was mainly related to the production of n-caproate and n-caprylate. Lactic acid bacteria of the genus Olsenella co-occurred with potentially chain-elongating bacteria of the genus Pseudoramibacter, and their abundance was positively correlated with n-caproate and n-caprylate concentrations. A new sub-network appeared in stage II, which was mainly related to n-butyrate production and revealed a network of different lactic acid bacteria (Bifidobacterium) and potential n-butyrate producers (Clostridium sensu stricto 12). The synergy effects between lactate-producing and lactate-consuming bacteria constitute a division of labor cooperation of mutual benefit. Besides cooperation, competition between different taxa determined the bacterial community assembly over the four succession stages in this resource-limited system. During long-term reactor operation under constant conditions, chain-elongating bacteria were outcompeted by butyrate-producing bacteria, leading to the increase of n-butyrate yield at the cost of medium-chain carboxylate yields in this closed model system.
Highlights
The production of platform chemicals and fuels from renewable resources is a major focus of a circular economy
The community dynamics analyzed by terminal restriction fragment length polymorphism (T-RFLP) fingerprinting is shown as Non-metric multidimensional scaling (NMDS) plots (Figure 2 for RsaI and Supplementary Figure S1 for MspI)
From 25 samples analyzed by amplicon sequencing, in total 909,240 sequence reads were obtained, which were assigned to 95 amplicon sequence variant (ASV) from high-quality sequence reads
Summary
The production of platform chemicals and fuels from renewable resources is a major focus of a circular economy. The carboxylate platform offers the opportunity to sustainably produce biobased chemicals such as medium-chain carboxylates (MCCs), which are mainly produced from coconut and palm kernel oils (Anneken et al, 2006). In a process known as chain elongation (CE), intermediates of acidogenesis such as acetate (C2) and n-butyrate (C4) can be elongated to MCCs by adding acetyl-CoA in reverse β-oxidation cycles (Spirito et al, 2014). C2 or C4 need to be transformed to acetyl-CoA or butyryl-CoA, respectively, as initial substrate for elongation in the reverse β-oxidation. Ethanol has been well described as electron donor providing energy for coupling acetyl-CoA formation and elongating acyl-CoA units (Seedorf et al, 2008). Feedstocks that are rich in lactate (e.g., ensiled plant biomass) or lactate-precursors (e.g., carbohydrates) are promising substrates for the production of MCCs
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