The nagB gene encoding beta-N-acetylglucosaminidase from S. thermoviolaceus OPC-520 was cloned and sequenced. The nagB gene could encode a protein of 541 amino acids with a calculated molecular mass of 58274. NagB revealed significant similarities to beta-N-acetylhexosaminidases and chitobiases from bacteria, which are classified into family 20 glycosyl hydrolases. NagB effectively hydrolyzed all of the chitin oligosaccharides from dimer to hexamer.