Naslov
Application of a mathematical model for the optimization of a multi-enzymatic reaction; The case study of L-homoserine synthesis

Autori
Česnik, Morana ; Sudar, Martina ; Hernandez, Karel ; Clapes, Pere ; Charnock, Simon ; Vasić-Rački, Đurđa ; Findrik Blažević, Zvjezdana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, stručni

Izvornik
5th multistep enzyme catalyzed processes congress : book of abstracts / Rother, Dörte ; Sehl, Torsten - Aachen : RWTH Aachen University, 2021, 26-26

Skup
5th Multistep Enzyme Catalyzed Processes Congress (MECP 2021)

Mjesto i datum
Online, 13.09.2021. - 16.09.2021

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
cascade reaction ; L-homoserine ; mathematical model ; optimization

Sažetak
Cascade multi-step reactions have gained a lot of attention in the last decade due to their numerous advantages against traditional organic synthesis methods. However, the number of dependencies between different variables increases with the number of enzymes and makes cascades highly complex to investigate, optimize and carry out. Consequently, for industrial application, it is important to understand their complexity from the viewpoint of enzyme reaction engineering. In this work the modelling methodology was applied on the synthesis of L-homoserine, which is an important intermediate and a building block in chemical and pharmaceutical industry. Its cascade synthesis catalyzed by an aldolase and transaminase in one pot starting from pyruvate, formaldehyde and L- alanine, was investigated in detail (Scheme 1), with enzymes, as well as whole lyophilized E. coli cells as catalysts. Scheme 1 The reaction scheme of the synthesis of L-homoserine. Reactions catalyzed by enzymes as cell free extracts (CFE) and E. coli cells containing the same co-expressed enzymes were optimized based on the developed mathematical model. The model was also used for the selection of the proper reactor set-up. Optimized reaction was carried out in the fed- batch reactor with continuous formaldehyde feed and two additions of L-alanine and pyruvate. The results were not only reproducible in our lab, but also in the lab of a project partner, and up- scaled experiments confirmed the applicability and potential of developed mathematical model for L- homoserine synthesis. The modeling approach enabled obtaining 672 mM of L-homoserine and volume productivity of 76.23 g L–1 d–1 after 25 hours in the experiment when CFE-enzymes were used as biocatalysts. Similar results were obtained in the experiment with lyophilized whole cells ; i.e. 640.74 mM of L- homoserine and volume productivity of 62 g L–1 d– 1. Obtained process metrics present an excellent result with a potential for an industrial use. Project CARBAZYMES-This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 635595.

Izvorni jezik
Engleski

Znanstvena područja
Kemijsko inženjerstvo, Biotehnologija

POVEZANOST RADA