Investigation of transaminase-based synthesis of furfurylamines
Time: Fri 2024-09-27 10.00
Location: Kollegiesalen, Brinellvägen 8, Stockholm
Video link: https://kth-se.zoom.us/j/63781939217
Language: English
Subject area: Biotechnology
Doctoral student: Luisa M. Merz , Industriell bioteknologi, Berglund Group
Opponent: Assoc. Prof. Ioannis Pavlidis, University of Crete, Grekland
Supervisor: Professor Per Berglund, Industriell bioteknologi; Dr. Luuk van Langen, ViaZym B.V.
QC 2024-09-02
Abstract
Biocatalysis presents a significant opportunity to enhance the sustainability of the chemical industry. Using enzymes for chemical production aligns closely with the principles of green chemistry, enabling a rapid, selective and low-waste synthesis of valuable compounds. Transaminases in particular offer the potential for selective, sustainable, and economical amine production. However, several challenges hinder their broader industrial application. This thesis investigates the transaminase from Silicibacter pomeroyi (SpATA) and how its applicability can be improved.
A critical challenge for transaminases is their stability, as the dimeric enzyme can readily dissociate, leading to a total loss of functionality. Various factors influencing the stability of SpATA were examined, demonstrating that buffer choice, cofactor concentration, and light exposure significantly impact enzyme stability. Optimising these conditions extended the half-life of SpATA to over 700 hours.
For large-scale applications, enzyme reusability is essential due to the significant cost of enzyme production. Therefore, SpATA, along with three other transaminases, was immobilised on a carrier, achieving excellent reusability without any loss of activity over five reaction cycles. Additionally, SpATA's application in a flow-reactor was successful, further demonstrating its industrial potential.
To enhance the sustainability of the transaminase reaction, the use of the “smart amine donor” cadaverine was investigated. High yields of the compound HMFA were achieved using equimolar amounts of cadaverine and HMF. To improve the economic viability of the process, an in-situ cascade reaction involving lysine decarboxylase and SpATA was developed for cadaverine production.
In summary, this thesis presents the development of a stable, reusable biocatalyst capable of achieving high product yields, significantly advancing the application of transaminases in sustainable industrial processes.