Advancing Biocatalysis with CYP450 A Computational Insight into Biofuel Production Water Dynamics and Ionic Liquid Effects

Abstract

In this thesis, we first attempted to investigate the fundamental structural aspect by which the enzyme CYP450OleT is catalytically unique in its family. The work shown in this Chapter resolves the ambiguity about the structural differences that control the decarboxylation reaction in CYP450OleT. Thereafter, we moved to computationally resolve the enigmatic decarboxylation in H85Q CYP450OleT , for which experimentalists have conflicting results. In a different chapter, we have explored the mysterious monooxygenase activity in the fusion of a redox protein with CYP450OleT in the formation of active oxidant Cpd I. In all the previous chapters, we have observed water molecules are important for the CYP450OleT enzyme to function, which excites us to benchmark the performance of computationally available water models (TIP3P, SPC/E, OPC) for different CYP450s. In a similar manner, we have further explored the different classes of solvents such as Ionic Liquids (ILs), and their effect on the stability and the properties of Cyt-C enzymes. In a nutshell, the present thesis unfolds various mysteries of CYP450OleT enzyme, which will be helpful for future metabolic engineering work towards better production of hydrocarbons for industrial applications. Benchmarking of different water models will be helpful for the accurate modelling of water in different enzymes. Moreover, the use of ionic liquid will provide a deep insight into the storage of environmentally sensitive Cyt-C enzymes.