Development of a thermostable β-glucuronidase-based reporter system for monitoring gene expression in hyperthermophiles

Mesophilic glucuronidases are the most widely used reporters of gene expression in plants, but unsuitable as reporters in (hyper-)thermophiles due their insufficient thermal stability. Here we present the native 66.8?kDa thermostable beta-glucuronidase of Sulfolobus solfataricus. The enzyme activity is characterized in a wide temperature range ideal for, but not limited to, in vivo genetic study of hyperthermophiles. As a proof of concept, we demonstrate its use as a reporter of gene expression in Sulfolobus, by monitoring a promoter fusion created with the beta-glucuronidase coding gene gusB and a copper-responsive promoter. Biotechnol. Bioeng. 2012; 109:18811886. (C) 2012 Wiley Periodicals, Inc.