Optimisation of halogenase enzyme activity by application of a genetic algorithm.
A genetic algorithm (GA) was applied for the optimisation of an enzyme assay composition respectively the enzyme activity of a recombinantly produced FADH(2)-dependent halogenating enzyme. The examined enzyme belongs to the class of halogenases and is capable to halogenate tryptophan regioselective in position 5. Therefore, the expressed trp-5-halogenase can be an interesting tool in the manufacturing of serotonin precursors. The application of stochastic search strategies (e.g. GAs) is well suited for fast determination of the global optimum in multidimensional search spaces, where statistical approaches or even the popular classical one-factor-at-a-time method often failures by misleading to local optima. The concentrations of six different medium components were optimised and the maximum yield of the halogenated tryptophan could be increased from 3.5 up to 65%.
Muffler
Kai
K
Institute of Bioprocess Engineering, TU Kaiserslautern, Gottlieb-Daimler-Strasse 44, D-67663 Kaiserslautern, Germany. muffler@rhrk.uni-kl.de
Retzlaff
Marco
M
van Pée
Karl-Heinz
KH
Ulber
Roland
R
D000465:Algorithms; D019020:Directed Molecular Evolution; D008956:Models, Chemical; D008957:Models, Genetic; D010088:Oxidoreductases / Q000737:chemistry* / Q000235:genetics*; D012701:Serotonin / Q000096:biosynthesis / Q000737:chemistry; D013379:Substrate Specificity; D014364:Tryptophan / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000737:chemistry
D012701:Serotonin; D014364:Tryptophan; D010088:Oxidoreductases; C407363:tryptophan halogenase