Constrained Allocation Flux Balance Analysis

New experimental results on bacterial growth inspire a novel top-down approach to study cell metabolism, combining mass balance and proteomic constraints to extend and complement Flux Balance Analysis. We introduce here Constrained Allocation Flux Balance Analysis, CAFBA, in which the biosynthetic costs associated to growth are accounted for in an effective way through a single additional genome-wide constraint. Its roots lie in the experimentally observed pattern of proteome allocation for metabolic functions, allowing to bridge regulation and metabolism in a transparent way under the principle of growth-rate maximization. We provide a simple method to solve CAFBA efficiently and propose an "ensemble averaging" procedure to account for unknown protein costs. Applying this approach to modeling E. coli metabolism, we find that, as the growth rate increases, CAFBA solutions cross over from respiratory, growth-yield maximizing states (preferred at slow growth) to fermentative states with carbon overflow (preferred at fast growth). In addition, CAFBA allows for quantitatively accurate predictions on the rate of acetate excretion and growth yield based on only 3 parameters determined by empirical growth laws.Comment: 21 pages, 6 figures (main) + 33 pages, various figures and tables (supporting); for the supplementary MatLab code, see http://tinyurl.com/h763es

Constrained Allocation Flux Balance Analysis

New experimental results on bacterial growth inspire a novel top-down approach to study cell metabolism, combining mass balance and proteomic constraints to extend and complement Flux Balance Analysis. We introduce here Constrained Allocation Flux Balance Analysis, CAFBA, in which the biosynthetic costs associated to growth are accounted for in an effective way through a single additional genome-wide constraint. Its roots lie in the experimentally observed pattern of proteome allocation for metabolic functions, allowing to bridge regulation and metabolism in a transparent way under the principle of growth-rate maximization. We provide a simple method to solve CAFBA efficiently and propose an “ensemble averaging” procedure to account for unknown protein costs. Applying this approach to modeling E. coli metabolism, we find that, as the growth rate increases, CAFBA solutions cross over from respiratory, growth-yield maximizing states (preferred at slow growth) to fermentative states with carbon overflow (preferred at fast growth). In addition, CAFBA allows for quantitatively accurate predictions on the rate of acetate excretion and growth yield based on only 3 parameters determined by empirical growth laws

Quantifying the entropic cost of cellular growth control

We quantify the amount of regulation required to control growth in living cells by a Maximum Entropy approach to the space of underlying metabolic states described by genome-scale models. Results obtained for E. coli and human cells are consistent with experiments and point to different regulatory strategies by which growth can be fostered or repressed. Moreover we explicitly connect the `inverse temperature' that controls MaxEnt distributions to the growth dynamics, showing that the initial size of a colony may be crucial in determining how an exponentially growing population organizes the phenotypic space.Comment: 3 page

Quantifying the benefit of a proteome reserve in fluctuating environments.

The overexpression of proteins is a major burden for fast-growing bacteria. Paradoxically, recent characterization of the proteome of Escherichia coli found many proteins expressed in excess of what appears to be optimal for exponential growth. Here, we quantitatively investigate the possibility that this overexpression constitutes a strategic reserve kept by starving cells to quickly meet demand upon sudden improvement in growth conditions. For cells exposed to repeated famine-and-feast cycles, we derive a simple relation between the duration of feast and the allocation of the ribosomal protein reserve to maximize the overall gain in biomass during the feast

Statistics of optimal information flow in ensembles of regulatory motifs

Genetic regulatory circuits universally cope with different sources of noise that limit their ability to coordinate input and output signals. In many cases, optimal regulatory performance can be thought to correspond to configurations of variables and parameters that maximize the mutual information between inputs and outputs. Such optima have been well characterized in several biologically relevant cases over the past decade. Here we use methods of statistical field theory to calculate the statistics of the maximal mutual information (the `capacity') achievable by tuning the input variable only in an ensemble of regulatory motifs, such that a single controller regulates N targets. Assuming (i) sufficiently large N, (ii) quenched random kinetic parameters, and (iii) small noise affecting the input-output channels, we can accurately reproduce numerical simulations both for the mean capacity and for the whole distribution. Our results provide insight into the inherent variability in effectiveness occurring in regulatory systems with heterogeneous kinetic parameters.Comment: 14 pages, 6 figure

Genome-scale models as a vehicle for knowledge transfer from microbial to mammalian cell systems

With the plethora of omics data becoming available for mammalian cell and, increasingly, human cell systems, Genome-scale metabolic models (GEMs) have emerged as a useful tool for their organisation and analysis. The systems biology community has developed an array of tools for the solution, interrogation and customisation of GEMs as well as algorithms that enable the design of cells with desired phenotypes based on the multi-omics information contained in these models. However, these tools have largely found application in microbial cells systems, which benefit from smaller model size and ease of experimentation. Herein, we discuss the major outstanding challenges in the use of GEMs as a vehicle for accurately analysing data for mammalian cell systems and transferring methodologies that would enable their use to design strains and processes. We provide insights on the opportunities and limitations of applying GEMs to human cell systems for advancing our understanding of health and disease. We further propose their integration with data-driven tools and their enrichment with cellular functions beyond metabolism, which would, in theory, more accurately describe how resources are allocated intracellularly

Advances in constraint-based models: methods for improved predictive power based on resource allocation constraints

The concept of metabolic models with resource allocation constraints has been around for over a decade and has clear advantages even when implementation is relatively rudimentary. Nonetheless, the number of organisms for which such a model is reconstructed is low. Various approaches exist, from coarse-grained consideration of enzyme usage to finegrained description of protein translation. These approaches are reviewed here, with a particular focus on user-friendly solutions that can introduce resource allocation constraints to metabolic models of any organism. The availability of kcat data is a major hurdle, where recent advances might help to fill in the numerous gaps that exist for this data, especially for nonmodel organisms

Microbial inefficient substrate use through the perspective of resource allocation models

Microorganisms extract energy from substrates following strategies that may seem suboptimal at first glance. Beyond the so-called yield-rate trade-off, resource allocation models, which focus on assigning different functional roles to the limited number of enzymes that a cell can support, offer a framework to interpret the inefficient substrate use by microorganisms. We review here relevant examples of substrate conversions where a significant part of the available energy is not utilised and how resource allocation models offer a mechanistic interpretation thereof, notably for open mixed cultures. Future developments are identified, in particular, the challenge of considering metabolic flexibility towards uncertain environmental changes instead of strict fixed optimality objectives, with the final goal of increasing the prediction capabilities of resource allocation models. Finally, we highlight the relevance of resource allocation to understand and enable a promising biorefinery platform revolving around lactate, which would increase the flexibility of waste-to-chemical biorefinery schemese authors would like to acknowledge the support of the Spanish Ministry of Education (FPU14/05457) and project CONSERVAL (INTERREG V-A Spain-Portugal, POCTEP), co-financed by the ERDF (Ref: 2352). The authors belong to the Galician Competitive Research Group (ED431C2017/029) and to the CRETUS Strategic Partnership (ED431E 2018/01), both programmes are co-funded by Xunta de Galicia and ERDF (EU)S

Universal scaling relation and criticality in metabolism and growth of Escherichia coli

The metabolic network plays a crucial role in regulating bacterial metabolism and growth, but it is subject to inherent molecular stochasticity. Previous studies have utilized flux balance analysis and the maximum entropy method to predict metabolic fluxes and growth rates, while the underlying principles governing bacterial metabolism and growth, especially the criticality hypothesis, remain unclear. In this study, we employ a maximum entropy approach to investigate the universality in various constraint-based metabolic networks of Escherichia coli. Our findings reveal the existence of universal scaling relations across different nutritional environments and metabolic network models, similar to the universality observed in physics. By analyzing single-cell data, we confirm that metabolism of Escherichia coli operates close to the state with maximum Fisher information, which serves as a signature of criticality. This critical state provides functional advantages such as high sensitivity and long-range correlation. Moreover, we demonstrate that a metabolic system operating at criticality takes a compromise solution between growth and adaptation, thereby serving as a survival strategy in fluctuating environments.Comment: 10 pages, 6 figure

Interrogating metabolism as an electron flow system

Metabolism is generally considered as a neatly organised system of modular pathways, shaped by evolution under selection for optimal cellular growth. This view falls short of explaining and predicting a number of key observations about the structure and dynamics of metabolism. We highlight these limitations of a pathway-centric view on metabolism and summarise studies suggesting how these could be overcome by viewing metabolism as a thermodynamically and kinetically constrained, dynamical flow system. Such a systems-level, first-principles based view of metabolism can open up new avenues of metabolic engineering and cures for metabolic diseases and allow better insights to a myriad of physiological processes that are ultimately linked to metabolism. Towards further developing this view, we call for a closer interaction among physical and biological disciplines and an increased use of electrochemical and biophysical approaches to interrogate cellular metabolism together with the microenvironment in which it exists