biostudies-literatureUnknown10(1)Fai Alex Wong CThe utilization of biocatalysts in biotechnological applications often necessitates their heterologous expression in suitable host organisms. However, the range of standardized microbial hosts for recombinant protein production remains limited, with most being mesophilic and suboptimal for certain protein types. Although the thermophilic bacterium <i>Thermus thermophilus</i> has long been established as a valuable extremophile host, thanks to its high-temperature tolerance, robust growth, and extensively characterized proteome, its genetic toolkit has predominantly depended on a limited set of native promoters. To overcome this bottleneck, we have expanded the available regulatory repertoire in <i>T. thermophilus</i> by developing novel artificial 5[Formula: see text] regulatory sequences Synthetic biology (Oxford, England)ysaf016https://www.ebi.ac.uk/biostudies/studies/S-EPMC12704435biostudies-literatureEngineering artificial 5' regulatory sequences for thermostable protein expression in the extremophile &lt;i&gt;Thermus thermophilus&lt;/i&gt;.PMC12704435Singh Dahiya GFai Alex Wong CLale RZhang STietze LfalseEngineering artificial 5' regulatory sequences for thermostable protein expression in the extremophile &lt;i&gt;Thermus thermophilus&lt;/i&gt;.The utilization of biocatalysts in biotechnological applications often necessitates their heterologous expression in suitable host organisms. However, the range of standardized microbial hosts for recombinant protein production remains limited, with most being mesophilic and suboptimal for certain protein types. Although the thermophilic bacterium <i>Thermus thermophilus</i> has long been established as a valuable extremophile host, thanks to its high-temperature tolerance, robust growth, and extensively characterized proteome, its genetic toolkit has predominantly depended on a limited set of native promoters. To overcome this bottleneck, we have expanded the available regulatory repertoire in <i>T. thermophilus</i> by developing novel artificial 5[Formula: see text] regulatory sequences 2025-01-01T00:00:00Z20252026-06-06T04:19:51.974Z2026-05-25T03:12:04.232ZS-EPMC127044354140442010.1093/synbio/ysaf016