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Lignocellulose forms plant cell walls, and its three constituent polymers, cellulose, hemicellulose and lignin, represent the largest renewable organic carbon pool in the terrestrial biosphere. Insights into biological lignocellulose deconstruction inform understandings of global carbon sequestration dynamics and provide inspiration for biotechnologies seeking to address the current climate crisis by producing renewable chemicals from plant biomass. Organisms in diverse environments disassemble lignocellulose, and carbohydrate degradation processes are well defined, but biological lignin deconstruction is described only in aerobic systems. It is currently unclear whether anaerobic lignin deconstruction is impossible because of biochemical constraints or, alternatively, has not yet been measured. We applied whole cell-wall nuclear magnetic resonance, gel-permeation chromatography and transcriptome sequencing to interrogate the apparent paradox that anaerobic fungi (Neocallimastigomycetes), well-documented lignocellulose degradation specialists, are unable to modify lignin. We find that Neocallimastigomycetes anaerobically break chemical bonds in grass and hardwood lignins, and we further associate upregulated gene products with the observed lignocellulose deconstruction. These findings alter perceptions of lignin deconstruction by anaerobes and provide opportunities to advance decarbonization biotechnologies that depend on depolymerizing lignocellulose.

While many heterologous molecules can be produced at trace concentrations via microbial bioconversion processes, maximizing their titers, rates, and yields from lignin-derived carbon streams remains challenging. Growth coupling can not only increase titers and yields but also shift the production period from stationary phase to growth phase. These methods for designing growth-coupling strains however require multi-gene edits for implementation which may be perceived as impractical. Here, we computationally evaluated 4,114 potential solutions for growth couplingpara-coumarate to indigoidine production and prototype two cut sets inPseudomonas putidaKT2440. We used adaptive laboratory evolution (ALE) on the initial triple deletion strain to restore growth onp-CA. Using X-ray tomography on this post-ALE strain we revealed increased cell density and decreased cell volume. Proteomics identified upregulated peroxidases that mitigate reactive oxygen species formation. Nine iterative stepwise modifications further informed by model-guided and rational approaches realized a growth coupled strain that produced 7.3 g/L indigoidine at 77% MTY inpara-coumarate minimal media. These ensemble strategies provide a blueprint for producing target molecules at high product titers, rates, and yields.

Standard procedures for evaluating environmental impact involve comparison between before and after conditions or scenarios or between treatment and control site pairs. In many cases, however, endogenous directional change (natural succession) is expected to occur at a significant rate over the period of concern, particularly for manmade systems such as impoundments. Static evaluations do not provide an adequate approach to such problems. A new evaluation frame is proposed. Nominal system behavior over time is characterized by a stochastic envelope around a nominal trajectory. We show that both the state variance and the sampling variance can change over time. In this context, environmental regulations can be framed as constraints, targets, or conformance to ideal trajectories. Statistical tests for determining noncompliance are explored relative to process variance, sample error, and sample size. Criteria are elucidated for choosing properties to monitor, sample size, and sampling interval.

Traditional fisheries management practices assume that increasing a minimum size limit will increase the sustained yield of larger fish by reducing the harvest of younger fish and increasing the recruitment into the older age groups. Simulation modeling of the northern pike (Esox lucius) population in the St. Lawrence River indicates that the yield of older fish actually declines under these conditions. In the absence of density-dependent compensation, the imposition of a 660-mm minimum size limit resulted in a greater decline in total population size than that predicted under a 508-mm limit over a 50-year period. A decrease in yield was caused by an increase in the harvest of older females and a subsequent decrease in egg production. To guard against possible population decline or shift in the age structure that reduces the stability and productivity of an existing fishery, it is suggested that reproductive potential be examined whenever minimum size limits are considered.