Galactokinase promiscuity: a question of flexibility?

Galactokinase catalyses the first committed step of the Leloir pathway, i.e. the ATP-dependent phosphorylation of α-D-galactose at C1-OH. Reduced galactokinase activity results in the inherited metabolic disease type II galactosaemia. However, inhibition of galactokinase is considered a viable approach to treating more severe forms of galactosaemia (types I and III). Considerable progress has been made in the identification of high affinity, selective inhibitors. Although the structure of galactokinase from a variety of species is known, its catalytic mechanism remains uncertain. Although the bulk of evidence suggests that the reaction proceeds via an active site base mechanism, some experimental and theoretical studies contradict this. The enzyme has potential as a biocatalyst in the production of sugar 1-phosphates. This potential is limited by its high specificity. A variety of approaches have been taken to identify galactokinase variants which are more promiscuous. These have broadened galactokinase's specificity to include a wide range of D- and L-sugars. Initial studies suggest that some of these alterations result in increased flexibility at the active site. It is suggested that modulation of protein flexibility is at least as important as structural modifications in determining the success or failure of enzyme engineering.

‘Harmonising the precaution out of the precautionary principle? The need for maintaining flexibility and an interpretative role’

The precautionary principle has the potential to act as a valuable tool in food law. It operates in areas of scientific uncertainty, calling for protective measures where there are potential threats to human health (or the environment). However, the manner of the principle’s incorporation and implementation within legislation are key to its effectiveness and general legitimacy. Specific considerations include the role and nature of risk assessments, assessors, sources of evidence, divergent opinions, risk communication, other legitimate factors and the weighting of interests. However, more fundamentally, the crystallisation of approaches and removal of all flexibility would undermine the principle’s central tenets. Firstly, principles crucially play a guiding and interpretative role. Secondly, reflexive modernisation and continuing scientific uncertainty call for the precautionary principle’s continued application – precautionary measures do not end the precautionary principle’s relevance. This can be partially achieved through the legislation so as to facilitate later precautionary measures, e.g. through temporary authorisations, derogations and safeguard clauses. However, crucially, it requires that the legislation also be interpreted in light of the precautionary principle. This paper investigates the logic behind the Court of Justice of the EU’s judgments and the circumstances that enable or deter the Court in taking, or permitting, stronger precautionary approaches. Although apparently inconsistent, a number of contextual factors including the legislative provisions and actors involved influence the judgments substantially. The analysis provides insight into improving the principle’s incorporation to facilitate its continued application and maintenance of flexibility, whilst bearing in mind the general desirability of objectivity and legal certainty.