Work

Engineering of an Evolved Artificial Formolase Enzyme To Facilitate In Vivo Synthetic C1 Metabolism

Year: 2025

Type: article

Abstract: High Resolution Image Download MS PowerPoint Slide To accomplish a sustainable circular economy, the conversion of CO 2 to products through biotechnological processes is a promising approach. One method uses synthetic methylotrophic microbes that use CO 2 -based methanol as the carbon source. However, its toxicity, as well as its byproduct formaldehyde, poses a challenge. To address this, a synthetic enzyme called formolase (FLS) has been designed to convert formaldehyde directly into dihydroxyacetone. So far, its utilization is limited due to low velocity, high K M, and the formation of the byproduct glycolaldehyde. To enable the in vivo application of formolase, the enzyme was subjected to iterative rounds of directed evolution. The key to success was a combined ″semirational” approach, which led to the identification of an advanced formolase variant exhibiting higher expression levels, enabling better formaldehyde tolerance in E. coli, and a 30% decrease in K M for the formose reaction. Furthermore, this double mutant exhibits an accelerated conversion of glycolaldehyde to dihydroxyacetone, which is particularly relevant for its future application in synthetic methylotrophy.

Source: ACS Sustainable Chemistry & Engineering

DOI: 10.1021/acssuschemeng.5c03359

Authors: Vanessa Wegat, Samed Güner, Manuel Döring, Jonathan T. Fabarius, Arne Roth, Volker Sieber

Institutions: Fraunhofer Institute for Interfacial Engineering and Biotechnology, Technical University of Munich

Language: English

Topic: Enzyme Catalysis and Immobilization

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Responsible consumption and production

Open Access status: hybrid

Estimated APC paid: $4,500