A 2-kb SmaI DNA fragment was cloned from the cosmid library of Streptomyces ansochromogenes. This DNA fragment contains a complete open reading frame which is 1275 bp in length, designated sanS (GenBank accession no. AF322179). The deduced SanS protein consists of 424 amino acids and belongs to a superfamily of enzymes with an unusual ATP-grasp fold. The disruption and complementation of sanS indicated that sanS is essential for nikkomycin biosynthesis in Streptomyces ansochromogenes. The sanS gene was subcloned into expression vector pET23b and overexpressed in E. coli BL21 (DE3). The protein was then purified and showed ATPase activity.