TY  - INPR
PB  - Wiley
Y1  - 2025/02/20/
A1  - Tonoli, A
A1  - Anselmi, S
A1  - Ward, JM
A1  - Hailes, HC
A1  - Jeffries, JWE
N2  - Transketolases (TKs) are thiamine diphosphate (ThDP)-dependent enzymes that catalyze the transfer of two-carbon units in a stereoselective manner, making them valuable biocatalysts for sustainable processes. Most known TKs are about 650 amino acids long; however, a second type found in Archaea and many Bacteria consists of two proteins, each of about 300 amino acids. Exploring the unique features and differences of split TKs may help in assessing their potential use in biocatalysis and for uncovering new reactivities. Additionally, it could provide valuable information on how their structure relates to their function, especially compared to full-length TKs. In this study, we significantly expanded the known repertoire of split TKs approximately 14-fold to the best of our knowledge, by identifying and providing accessions of nearly 500 putative split-TK subunit pairs. Moreover, we doubled the number of experimentally produced and tested split TKs by cloning, purifying, and testing ten candidates retrieved from genomes and in-house metagenomes. Interestingly, pQR2809 and pQR2812, derived from hyperthermophilic organisms, showed enhanced thermostability compared to other TK examples in the literature, maintaining partial activity after heating at 90 °C or 100 °C for 1 hour, respectively.
AV  - public
JF  - ChemBioChem
SN  - 1439-4227
N1  - © 2025 The Author(s). ChemBioChem published by Wiley-VCH GmbH

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
ID  - discovery10205686
TI  - Expanding the Enzymatic Toolbox for Carboligation: Increasing the Diversity of the ?Split? Transketolase Sequence Space
KW  - Biocatalysis
KW  -  Enzymes
KW  -  Metagenomics
KW  -  Thermostability
KW  -  Transketolase
UR  - https://doi.org/10.1002/cbic.202401028
ER  -