Amine transaminases (ATAs), belonging to the class III transaminases within the superfamily of pyridoxal-5 '-phosphate-dependent enzymes, catalyze transamination reactions between amino donors and amino acceptors. These enzymes are particularly appealing for their role in stereospecific synthesis of chiral amines. However, the stability of most ATAs is not satisfying, limiting their suitability for industrial applications. Among them, the amine transaminase from Silicibacter pomeroyi (Sp-ATA) has drawn attention due to its high activity and broad substrate scope under mild conditions and high pH. Nevertheless, maintaining the activity at higher temperatures is a challenge. Previous studies to enhance enzyme function through directed evolution have shown promising results, yet predicting the cooperative effects of individual stabilizing mutations remains challenging. An alternative strategy is ancestral sequence reconstruction (ASR), which is based on gene sequences to create a more or less artificial phylogenetic tree. This study aims to leverage ASR techniques to explore the thermostability, solvent tolerance, and substrate profile of Sp-ATA, to find more stable transaminases. By using Sp-ATA as a template and incorporating insights from ancestral sequences, this strategy offers a promising approach for developing robust biocatalysts suitable for industrial applications.

The fashion industry is becoming one of the largest emitters worldwide due to its high consumption of raw materials, its effluents, and the fact that every garment will eventually contribute to the vast amount of waste being incinerated or accumulating in landfills. Although fiber-to-fiber recycling processes are being developed, the mechanical properties of the textile fibers are typically degraded with each such recycle. Thus, tertiary recycling alternatives where textiles are depolymerized to convert them into valuable products are needed to provide end-of-life alternatives and to achieve circularity in the fashion industry. We have developed a method whereby cotton waste textiles are depolymerized to form a glucose solution, using sulfuric acid as the sole catalyst, with a high yield (>70%). The glucose solution produced in this process has a high concentration (>100 g/L), which reduces the purification cost and makes the process industrially relevant. This method can be applied regardless of the quality of the fibers and could therefore process other cellulosic fibers such as viscose. The glucose produced could subsequently be fermented into butanediol or caprolactam, precursors for the production of synthetic textile fibers, thus retaining the value of the waste textiles within the textile value chain.