The analysis of the dynamic behaviour of genome-scale models of metabolism (GEMs) currently presents considerable challenges because of the difficulties of simulating such large and complex networks. Bacterial GEMs can comprise about 5000 reactions and metabolites, and encode a huge variety of growth conditions; such models cannot be used without sophisticated tool support. This article is intended to aid modellers, both specialist and non-specialist in computerized methods, to identify and apply a suitable combination of tools for the dynamic behaviour analysis of large-scale metabolic designs. We describe a methodology and related workflow based on publicly available tools to profile and analyse whole-genome-scale biochemical models. We use an efficient approximative stochastic simulation method to overcome problems associated with the dynamic simulation of GEMs. In addition, we apply simulative model checking using temporal logic property libraries, clustering and data analysis, over time series of reaction rates and metabolite concentrations. We extend this to consider the evolution of reaction-oriented properties of subnets over time, including dead subnets and functional subsystems. This enables the generation of abstract views of the behaviour of these models, which can be large-up to whole genome in size-and therefore impractical to analyse informally by eye. We demonstrate our methodology by applying it to a reduced model of the whole-genome metabolism of Escherichia coli K-12 under different growth conditions. The overall context of our work is in the area of model-based design methods for metabolic engineering and synthetic biology.

Systems and synthetic biology require multiscale biomodel engineering approaches to integrate diverse spatial and temporal scales which will help to better understand and describe the various interactions in biological systems. Our BioModelKit framework for modular biomodel engineering allows compo sing multiscale models from a set of modules, each describing an individual biomolecular component in the form of a Petri net. In this framework, we now propose a feature for spatial modelling of molecular biosystems. The spatial model represents the coordinates and movement of individual biomolecular components on a defined grid. Here, the distance between components constrains their ability to interact with each other. We use coloured Petri nets to scale the spatial model, such that each molecular component can exist in an arbitrary number of instances. The grid can encode various regular and irregular structures according to the cellular arrangement and geometry. Furthermore, the grid can be divided into compartments to represent the compartmentalisation of the cell.

We present an approach to improve the efficiency of stochastic simulation for large and dense biochemical reaction networks. We use stochastic Petri nets as modelling framework, but the proposed simulation approach is not limited to Petri net representations. The underlying continuous-time Markov c hain (CTMC) is converted to an equivalent discrete-time Markov chain (DTMC); this itself gains no efficiency. We improve the efficiency via discrete-time leaps, even though this results in an approximate method. The discrete-time leaps are done by applying the maximum firing rule; this reduces drastically the number of steps. The presented algorithm is implemented in our modelling and simulation tool Snoopy, as well as in our advanced analysis and model checking tool MARCIE. We demonstrate the approach on models of different sizes and complexities.

We consider localised DNA computation, where a DNA strand walks along a binary decision graph to compute a binary function. One of the challenges for the design of reliable walker circuits consists in leakage transitions, which occur when a walker jumps into another branch of the decision graph. We automatically identify leakage transitions, which allows for a detailed qualitative and quantitative assessment of circuit designs, design comparison, and design optimisation. The ability to identify leakage transitions is an important step in the process of optimising DNA circuit layouts where the aim is to minimise the computational error inherent in a circuit while minimising the area of the circuit. Our 2D modelling approach of DNA walker circuits relies on coloured stochastic Petri nets which enable functionality, topology and dimensionality all to be integrated in one two-dimensional model. Our modelling and analysis approach can be easily extended to 3-dimensional walker systems.

SUMMARY: To investigate biomolecular networks, Snoopy provides a unifying Petri net framework comprising a family of related Petri net classes. Models can be hierarchically structured, allowing for the mastering of larger networks. To move easily between the qualitative, stochastic and continuous modelling paradigms, models can be converted into each other. We get models sharing structure, but specialized by their kinetic information. The analysis and iterative reverse engineering of biomolecular networks is supported by the simultaneous use of several Petri net classes, while the graphical user interface adapts dynamically to the active one. Built-in animation and simulation are complemented by exports to various analysis tools. Snoopy facilitates the addition of new Petri net classes thanks to its generic design. AVAILABILITY: Our tool with Petri net samples is available free of charge for non-commercial use at http://www-dssz.informatik.tu-cottbus.de/snoopy.html; supported operating systems: Mac OS X, Windows and Linux (selected distributions).