Rational conversion of substrate and product specificity in a Salvia monoterpene synthase: structural insights into the evolution of terpene synthase function

Terpene synthases are responsible for the biosynthesis of the complex chemical defense arsenal of plants and microorganisms. How do these enzymes, which all appear to share a common terpene synthase fold, specify the many different products made almost entirely from one of only three substrates? Elucidation of the structure of 1,8-cineole synthase from Salvia fruticosa (Sf-CinS1) combined with analysis of functional and phylogenetic relationships of enzymes within Salvia species identified active-site residues responsible for product specificity. Thus, Sf-CinS1 was successfully converted to a sabinene synthase with a minimum number of rationally predicted substitutions, while identification of the Asn side chain essential for water activation introduced 1,8-cineole and alpha-terpineol activity to Salvia pomifera sabinene synthase. A major contribution to product specificity in Sf-CinS1 appears to come from a local deformation within one of the helices forming the active site. This deformation is observed in all other mono- or sesquiterpene structures available, pointing to a conserved mechanism. Moreover, a single amino acid substitution enlarged the active-site cavity enough to accommodate the larger farnesyl pyrophosphate substrate and led to the efficient synthesis of sesquiterpenes, while alternate single substitutions of this critical amino acid yielded five additional terpene synthases.

The capability-affordance model: a method for analysis and modelling of capabilities and affordances

Existing capability models lack qualitative and quantitative means to compare business capabilities. This paper extends previous work and uses affordance theories to consistently model and analyse capabilities. We use the concept of objective and subjective affordances to model capability as a tuple of a set of resource affordance system mechanisms and action paths, dependent on one or more critical affordance factors. We identify an affordance chain of subjective affordances by which affordances work together to enable an action and an affordance path that links action affordances to create a capability system. We define the mechanism and path underlying capability. We show how affordance modelling notation, AMN, can represent affordances comprising a capability. We propose a method to quantitatively and qualitatively compare capabilities using efficiency, effectiveness and quality metrics. The method is demonstrated by a medical example comparing the capability of syringe and needless anaesthetic systems.