Applied Microbiology and Biotechnology, Vol.93, No.6, 2403-2410, 2012
A thermostable recombinant transaldolase with high activity over a broad pH range
Thermophilic enzymes are in high demand for various applications due to their prolonged lifetimes and high reaction rates at elevated temperatures. In this work, an open reading frame TM0295, which encodes a putative transaldolase (TAL) from a hyper-thermophilic microorganism, Thermotoga maritima, was cloned and expressed in Escherichia coli. The enzyme activity of transaldolase at high temperatures (e.g., at 80 A degrees C) was reported here for the first time. The recombinant T. maritima transaldolase was extremely thermostable, with a half-life time of 198 and 13.0 h at 60 A degrees C and 80 A degrees C, respectively. The estimated total turn-over number was 1.5 x 10(6) mol of product per mol of enzyme at 80 A degrees C. This enzyme also exhibited high activities within a broad pH range of 6.0-9.0. This ultra-thermostable TAL with high activity shows great potential for use in such applications as the production of enzymatic biofuels production and the synthesis of high-value carbohydrates by cell-free synthetic pathway biotransformation.
Keywords:Building block;Cell-free synthetic biology;Transaldolase activity;Thermotoga maritima;Thermostability