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β-N-Acetylglucosaminidase has been purified from the walls of Bacillus subtilis 168 and compared with the other known autolysin, N-acetylmuramyl-l-alanine amidase (amidase). The β-N-acetylglucosaminidase was a dimer in LiCl buffers with a sub-unit molecular weight of 90000 and a pH optimum of about 5·0. It was very sensitive to proteolytic enzymes and was critically activated by 0·1 to 0·2 m-LiCl. It was insoluble in concentrations of LiCl lower than 0·05 to 0·1 m. It was less strongly bound to walls than was the amidase, which was a monomer of molecular weight 30000 to 40000 in LiCl buffers. The βN-acetylglucosaminidase is an endoenzyme and showed no exo-activity. Lysozyme-like enzyme (muramidase) activity was undetectable in the wall extracts examined.
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